1.Screening of the specific aptamer of human CD20 extracellular protein expressed in Escherichia coli by systematic evolution of ligands by exponential enrichment.
Fan CHEN ; Fan YANG ; Lei GAO ; Yue HU ; Yun XUE ; Jing ZHOU ; Jianhua KANG ; Wei WANG
Chinese Journal of Biotechnology 2025;41(4):1467-1477
CD20 is a surface marker protein of B-cell lymphoma, and its extracellular region is the target of specific antibodies and drugs. To obtain a cheap and easily modified specific preparation targeting CD20, we optimized the gene of CD20 extracellular region according to codon degeneracy to facilitate its expression in Escherichia coli. The optimized gene was cloned into pGEX-4T-1 vector, and the recombinant vector was transformed into E. coli BL21(DE3) for expression. The purified protein was identified by SDS-PAGE and Western blotting. Systematic evolution of ligands by exponential enrichment (SELEX) was employed to screen the ssDNA aptamer that specifically binds to the fusion protein, and the affinity of the aptamer to CD20 was detected by flow cytometry. Then, the cytotoxicity test was carried out to examine the inhibitory effect of the aptamer on B lymphoma cells. In this study, we established the prokaryotic expression method of CD20 and obtained the aptamer specifically binding to the extracellular region of CD20, which laid a foundation for the development of therapeutic drugs targeting CD20.
Humans
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Escherichia coli/metabolism*
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SELEX Aptamer Technique/methods*
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Aptamers, Nucleotide/genetics*
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Antigens, CD20/metabolism*
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Ligands
2.Influence of muscle activation on lumbar injury under a specific +Gz load
Kang LEI ; Chen-Zhu ZHAO ; Fan LI ; Shu-Lin LIU ; Peng YANG
Chinese Journal of Traumatology 2024;27(4):218-225
Purpose::The present study aimed to analyze the influence of muscle activation on lumbar injury under a specific +Gz load.Methods::A hybrid finite element human body model with detailed lumbar anatomy and lumbar muscle activation capabilities was developed. Using the specific +Gz loading acceleration as input, the kinematic and biomechanical responses of the occupant's lower back were studied for both activated and deactivated states of the lumbar muscles.Results::The results indicated that activating the major lumbar muscles enhanced the stability of the occupant's torso, which delayed the contact between the occupant's head and the headrest. Lumbar muscle activation led to higher strain and stress output in the lumbar spine under +Gz load, such as the maximum Von Mises stress of the vertebrae and intervertebral discs increased by 177.9% and 161.8%, respectively, and the damage response index increased by 84.5%.Conclusion::In both simulations, the occupant's risk of lumbar injury does not exceed 10% probability. Therefore, the activation of muscles could provide good protection for maintaining the lumbar spine and reduce the effect of acceleration in vehicle travel direction.
3.Interpretation of consensus guidelines for the diagnosis and management of succinic semialdehyde dehydroge-nase deficiency formulated by the international SSADHD consensus group in 2024
Beibei KANG ; Lei XU ; Qiang YU ; Yanping FAN ; Lijie ZHU ; Xuemei ZHENG ; Jianguo CAO ; Jiaqin WANG
Chinese Journal of Applied Clinical Pediatrics 2024;39(10):738-742
Succinic semialdehyde dehydrogenase deficiency (SSADHD) is a rare autosomal recessive neurometabolic disease.Pathogenic mutations in ALDH5A1 genes lead to abnormalities in the structure, activity and function of succinic semialdehyde dehydrogenase, resulting in a series of neurological damage.Due to the rarity of SSADHD and the huge differences in its clinical manifestations, it often leads to misdiagnosis or delayed diagnosis, and the treatment is mainly symptomatic.There is no specific drug or treatment.In March 2024, the SSADHD consensus group, composed of SSADHD researchers from 19 institutions in 11 countries and regions, released the " Consensus Guidelines for the Diagnosis and Management of Succinic Semialdehyde Dehydrogenase Deficiency" , which elaborates on the definition, epidemiology, clinical manifestations, diagnosis, and treatment of SSADHD, aiming to standardize and unify the diagnosis and management of SSADHD.This article interprets the key contents of the guidelines, in order to provide guidance for the early screening, diagnosis and treatment of SSADHD in China.
4.Research status of non-coding RNA in viral myocarditis
Xiao-Long HE ; Xin-Xin HU ; Fan-Ning WANG ; Wen-Xin WANG ; Guo-Lei ZHOU ; Kang YI ; Tao YOU
The Chinese Journal of Clinical Pharmacology 2024;40(14):2143-2147
Viral myocarditis(VMC)is the leading cause of dilated cardiomyopathy,which can lead to heart failure and sudden cardiac death.With the development of high-throughput sequencing technology,non-coding RNA(ncRNA)plays an important role in the occurrence and development of VMC.ncRNA promotes the occurrence and development of VMC by regulating viral replication,immune cell function,myocardial cell death,myocardial interstitial fibrosis,and other pathological processes.This article reviews the research progress of ncRNA in VMC and provides new ideas for the pathogenesis,diagnosis,and treatment of VMC.
5.A prospective study on the association between lifestyles and mortality risk in adults in Henan Province
Lei FAN ; Minjie QI ; Tianfang XING ; Gang HOU ; Hanxue ZHANG ; Sen LIANG ; Li HAN ; Wenxie DING ; Kai KANG ; Zhiwei HAN
Chinese Journal of Epidemiology 2024;45(8):1052-1058
Objective:To analyze the association between healthy lifestyle and mortality among Henan Province 35-74 years old individuals.Methods:Data from the programme of screening and intervention subjects with high-risk cardiovascular disease 99 133 adults were analyzed in a provincial cohort study of 16 counties. Four healthy lifestyle behaviors were assessed based on a questionnaire survey. Information on mortality endpoints was retrieved from the national death surveillance system. Cox proportional hazards regression models were used to estimate the associations between healthy lifestyles, mortality risk and population attributable fraction (PAF).Results:Out of the adult participants in Henan, 50.6% adhered to a healthy lifestyle, and only 0.1% adhered to 4 healthy lifestyle behaviours. During a mean of 4.5 years, 2 685 all-cause death and 1 283 cardiovascular deaths were documented. The decreased risk of mortality among individuals with non-smoking, moderate drinking, adequate exercise and healthy diet were 0.85 (95% CI: 0.77-0.94), 0.75 (95% CI: 0.63-0.89), 0.73 (95% CI: 0.67-0.79) and 0.86 (95% CI: 0.77-0.96), while the adjusted PAF for all-cause deaths were 5.2% (95% CI: 2.5%-7.9%), 24.0% (95% CI: 10.7%-36.4%), 19.4% (95% CI: 13.8%-24.8%) and 12.3% (95% CI: 3.4%-20.9%), respectively. A combined healthy lifestyle can bring more health benefits. Adherence to 4 healthy lifestyle behaviours could avoid 49.1% of all-cause death. Conclusion:Adherence to a healthy lifestyle can reduce the risk of death, and participants with a healthy lifestyle had a lower mortality risk.
6.Safety and efficacy of neoadjuvant chemotherapy combined with immunotherapy in 101 patients with muscle-invasive bladder cancer
Chaosheng GAN ; Tao LI ; Junjie FAN ; Zhangdong JIANG ; Guojing WANG ; Ke XU ; Qiyuan KANG ; Yangqingqing ZHOU ; Yuefeng DU ; Jinhai FAN ; Lei LI ; Dalin HE ; Kaijie WU
Journal of Modern Urology 2024;29(9):790-796
Objective To explore the safety and efficacy of neoadjuvant chemotherapy(NAC)combined with immunotherapy before radical cystectomy plus pelvic lymph nodes dissection(RC-PLND)for muscle-invasive bladder cancer(MIBC).Methods The clinical data of 101 patients with MIBC who underwent neoadjuvant therapy followed by RC-PLND in the Department of Urology,the First Affiliated Hospital of Xi'an Jiaotong University during Jan.2019 and Dec.2023 were retrospectively analyzed,including 71 patients(70.3%)who received NAC(NAC group)and 30(29.7%)who received NAC combined with immunotherapy(NAC combine immunotherapy group).The clinical and pathological data and adverse events during neoadjuvant therapy were compared.Logistic regression analysis was used to explore the independent predictors of pathological complete response(pCR)and pathological partial response(pPR).Results There were no significant differences in the baseline data between the two groups(P>0.05).However,the proportion of multiple tumors in patients receiving NAC before surgery was significantly higher than that in the NAC combined immunotherapy group(69.0%vs.46.7%,P=0.034).Compared with NAC group,NAC combined with immunotherapy group had significantly improved rate of pathological downstaging and pPR(60.6%vs.83.3%,P=0.026;45.1%vs.70.0%,P=0.022).Furthermore,the rate of pCR in patients undergoing NAC combined immunotherapy was higher than those undergoing NAC,but the difference was not significant(53.3%vs.33.8%,P=0.067).Logistic regression analysis revealed that clinical T-stage and tumor diameter were independent predictors of pCR and pPR(P<0.05).In addition,the most common adverse events during neoadjuvant therapy were anemia,decreased white blood cells,nausea,and vomiting,but most of them were grade 1-2 and could be relieved through symptomatic treatment.Conclusion NAC combined with immunotherapy is safe and effective,which can improve the rate of pathological downstaging,pPR and pCR,without increasing the incidence of adverse reactions.
7.Impact of ultrasound-guided thoracic paravertebral nerve block combined with general anesthesia in patients undergoing thoracoscopic surgery and its potential mechanisms
Xiaomeng ZHU ; Sa FAN ; Lei KANG ; Yilan ZHANG ; Chao YUAN ; Fei HUANG
Journal of Clinical Medicine in Practice 2024;28(14):44-48
Objective To investigate the effects and potential mechanisms of ultrasound-guided thoracic paravertebral block (TPVB) combined with general anesthesia in patients undergoing thoracoscopic surgery. Methods A total of 122 patients undergoing thoracoscopic surgery were selected and randomly divided into control group and tudy group, with 61 patients in each group.The control group received general anesthesia, while the study group received ultrasound-guided TPVB combined with general anesthesia.Clinical data and perioperative indicators were recorded and compared between the two groups.Mean arterial pressure (MAP), heart rate (HR), Visual Analogue Scale (VAS) scores before surgery, 24 hours and 72 hours after surgery, and inflammatory factors[interleukin-6(IL-6), interleukin-10(IL-10), tumor necrosis factor-α(TNF-α)], oxidative stress indicators[malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px)]levels before surgery and 72 hours after surgery were compared between the two groups.Postoperative adverse reactions were recorded.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of key molecules mRNA in the Nrf2/Keap1/ARE pathway in the peripheral blood of patients in the two groups. Results There were no statistically significant differences in operative time and intraoperative blood loss between the two groups (
8.A third dose of inactivated vaccine augments the potency, breadth, and duration of anamnestic responses against SARS-CoV-2.
Zijing JIA ; Kang WANG ; Minxiang XIE ; Jiajing WU ; Yaling HU ; Yunjiao ZHOU ; Ayijiang YISIMAYI ; Wangjun FU ; Lei WANG ; Pan LIU ; Kaiyue FAN ; Ruihong CHEN ; Lin WANG ; Jing LI ; Yao WANG ; Xiaoqin GE ; Qianqian ZHANG ; Jianbo WU ; Nan WANG ; Wei WU ; Yidan GAO ; Jingyun MIAO ; Yinan JIANG ; Lili QIN ; Ling ZHU ; Weijin HUANG ; Yanjun ZHANG ; Huan ZHANG ; Baisheng LI ; Qiang GAO ; Xiaoliang Sunney XIE ; Youchun WANG ; Yunlong CAO ; Qiao WANG ; Xiangxi WANG
Protein & Cell 2024;15(12):930-937
9.The effect of on-demand glucocorticoid strategy on the occurrence and outcome of p-ALG-associated serum sickness in aplastic anemia
Xiawan YANG ; Kang ZHOU ; Jianping LI ; Huihui FAN ; Wenrui YANG ; Lei YE ; Yuan LI ; Yang LI ; Guangxin PENG ; Yang YANG ; Youzhen XIONG ; Xin ZHAO ; Liping JING ; Li ZHANG ; Fengkui ZHANG
Chinese Journal of Hematology 2023;44(3):211-215
Objective:To investigate the effect of on-demand glucocorticoid strategy on the occurrence and outcome of porcine anti-lymphocyte globulin (p-ALG) -associated serum sickness in aplastic anemia (AA) .Methods:The data of AA patients who received in the Anemia Diagnosis and Treatment Center of Haematology Hospital, CAMS & PUMC from January 2019 to January 2022 were collected. Among them, 35 patients were enrolled in the on-demand group, with the glucocorticoid strategy adjusted based on the occurrence and severity of serum sickness; 105 patients were recruited in the usual group by matching the age and disease diagnosis according to 1∶3 ratio in patients who received a conventional glucocorticoid strategy in the same period. The incidences, clinical manifestations, treatment outcomes of serum sickness, and glucocorticoid dosage between the two groups were analyzed.Results:The incidences of serum sickness in the on-demand group and the usual group were 65.7% and 54.3% ( P=0.237) , respectively. The median onset of serum sickness was the same [12 (9, 13) d vs the 12 (10, 13) d, P=0.552], and clinical symptoms and signs, primarily joint, and/or muscle pain, fever, and rash were similar. Severity grades were both dominated by Grades 1-2 (62.8% vs 51.4%) , with only a few Grade 3 (2.9% vs 2.9%) , and no Grades 4-5. No significant difference in the serum sickness distribution ( P=0.530) . The median duration of serum sickness was the same [5 (3, 7) d vs 5 (3, 6) d, P=0.529], and all patients were completely cured after glucocorticoid therapy. In patients without serum sickness, the average dosage of prophylactic glucocorticoid per patient in the usual group was (469.48 ±193.57) mg (0 in the on-demand group) . When compared to the usual group, the average therapeutic glucocorticoid dosage per patient in the on-demand group was significantly lower [ (125.91±77.70) mg vs (653.90±285.56) mg, P<0.001]. Conclusions:In comparison to the usual glucocorticoid strategy, the on-demand treatment strategy could significantly reduce glucocorticoid dosage without increasing the incidence of serum sickness; in addition, the duration of serum sickness and the incidence of above Grade 2-serum sickness were similar.
10.Metagenomic next-generation sequencing of plasma for the identification of bloodstream infectious pathogens in severe aplastic anemia
Yuan LI ; Youzhen XIONG ; Huihui FAN ; Liping JING ; Jianping LI ; Qingsong LIN ; Chunhui XU ; Ying LI ; Lei YE ; Meng JIAO ; Yang YANG ; Yang LI ; Wenrui YANG ; Guangxin PENG ; Kang ZHOU ; Xin ZHAO ; Li ZHANG ; Fengkui ZHANG
Chinese Journal of Hematology 2023;44(3):236-241
Objective:To analyze the diagnostic value of cell-free plasma metagenomic next-generation sequencing (mNGS) pathogen identification for severe aplastic anemia (SAA) bloodstream infection.Methods:From February 2021 to February 2022, mNGS and conventional detection methods (blood culture, etc.) were used to detect 33 samples from 29 consecutive AA patients admitted to the Anemia Diagnosis and Treatment Center of the Hematology Hospital of the Chinese Academy of Medical Sciences to assess the diagnostic consistency of mNGS and conventional detection, as well as the impact on clinical treatment benefits and clinical accuracy.Results:①Among the 33 samples evaluated by mNGS and conventional detection methods, 25 cases (75.76%) carried potential pathogenic microorganisms. A total of 72 pathogenic microorganisms were identified from all cases, of which 65 (90.28%) were detected only by mNGS. ②All 33 cases were evaluated for diagnostic consistency, of which 2 cases (6.06%) were Composite, 18 cases (54.55%) were mNGS only, 2 cases (6.06%) were Conventional method only, 1 case (3.03%) was both common compliances (mNGS/Conventional testing) , and 10 cases (30.3%) were completely non-conforming (None) . ③All 33 cases were evaluated for clinical treatment benefit. Among them, 8 cases (24.24%) received Initiation of targeted treatment, 1 case (3.03%) received Treatment de-escalation, 13 cases (39.39%) received Confirmation, and the remaining 11 cases (33.33%) received No clinical benefit. ④ The sensitivity of 80.77%, specificity of 70.00%, positive predictive value of 63.64%, negative predictive value of 84.85%, positive likelihood ratio of 2.692, and negative likelihood ratio of 0.275 distinguished mNGS from conventional detection methods (21/12 vs 5/28, P<0.001) . Conclusion:mNGS can not only contribute to accurately diagnosing bloodstream infection in patients with aplastic anemia, but can also help to guide accurate anti-infection treatment, and the clinical accuracy is high.


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