1.Recombinant expression of Sphingobium yanoikuyae esterase SyEst870 capable of degrading carbamate pesticides.
Xiaoqian XIE ; Yin FENG ; Yuanyuan ZHOU ; Xin YAN ; Xiaoqin YUAN ; Wuxia QIU ; Xinfang MAO ; Zhongyuan LIU
Chinese Journal of Biotechnology 2025;41(4):1605-1620
Carbamate pesticides, a new type of broad-spectrum pesticides for controlling pests, mites, and weeds, are developed to address the shortcomings of organochlorine and organophosphorus pesticides. Their widespread use and slow degradation have led to environmental pollution, causing damage to ecosystems and human health. Managing pesticide residues is a pressing issue in the current environmental protection. This study aims to investigate the expression of SyEst870, a member of the SGNH/GDSL hydrolase family in Sphingobium yanoikuyae, in a prokaryotic system and evaluate the ability of the recombinant protein to degrade carbamate pesticides. The prokaryotic expression vector pET-32a-SyEst870 was constructed and transformed into the Escherichia coli BL21 for heterologous expression. The purified protein was studied in terms of enzyme activity and effects of temperature, pH, and metal ions on the enzyme activity, with p-nitrophenol acetate as the substrate and based on the standard curve of p-nitrophenol. LC-MS (liquid chromatography-mass spectrometry) was employed to examine the degradation effects of SyEst870 on carbaryl, metolcarb, and isoprocarb. GC-MS (gas chromatography-mass spectrometry) was employed to detect the degradation products of SyEst870 for the three pesticides. The soluble protein SyEst870 was successfully obtained through the heterologous expression in Escherichia coli, which yielded an enzyme with the activity of 677.5 U after affinity chromatography. SyEst870 exhibited degradation rates of 82.34%, 84.43%, and 92.87% for carbaryl, metolcarb, and isoprocarb, respectively, at an initial concentration of 100 mg/L within 24 h at 30 ℃ and pH 7.0. The primary degradation products of carbaryl were identified as α-naphthol and methyl isocyanate. Metolcarb was mainly degraded into m-cresol and methyl isocyanate, and isoprocarb was mainly degraded into 2-isopropylphenol and methyl isocyanate. Compared with the half-life of carbamate pesticides in the natural environment, which ranges from a few days to several weeks, the recombinant protein SyEst870 can rapidly eliminate the residues of carbamate pesticides. This study lays a foundation for addressing pesticide residues in the environment and in fruits and vegetables.
Escherichia coli/metabolism*
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Sphingomonadaceae/genetics*
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Recombinant Proteins/metabolism*
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Biodegradation, Environmental
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Esterases/metabolism*
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Pesticides/isolation & purification*
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Carbamates/isolation & purification*
2.Single-Cell Mapping of Brain Myeloid Cell Subsets Reveals Key Transcriptomic Changes Favoring Neuroplasticity after Ischemic Stroke.
Fangxi LIU ; Xi CHENG ; Chuansheng ZHAO ; Xiaoqian ZHANG ; Chang LIU ; Shanshan ZHONG ; Zhouyang LIU ; Xinyu LIN ; Wei QIU ; Xiuchun ZHANG
Neuroscience Bulletin 2024;40(1):65-78
Interactions between brain-resident and peripheral infiltrated immune cells are thought to contribute to neuroplasticity after cerebral ischemia. However, conventional bulk sequencing makes it challenging to depict this complex immune network. Using single-cell RNA sequencing, we mapped compositional and transcriptional features of peri-infarct immune cells. Microglia were the predominant cell type in the peri-infarct region, displaying a more diverse activation pattern than the typical pro- and anti-inflammatory state, with axon tract-associated microglia (ATMs) being associated with neuronal regeneration. Trajectory inference suggested that infiltrated monocyte-derived macrophages (MDMs) exhibited a gradual fate trajectory transition to activated MDMs. Inter-cellular crosstalk between MDMs and microglia orchestrated anti-inflammatory and repair-promoting microglia phenotypes and promoted post-stroke neurogenesis, with SOX2 and related Akt/CREB signaling as the underlying mechanisms. This description of the brain's immune landscape and its relationship with neurogenesis provides new insight into promoting neural repair by regulating neuroinflammatory responses.
Humans
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Ischemic Stroke
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Brain/metabolism*
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Macrophages
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Brain Ischemia/metabolism*
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Microglia/metabolism*
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Gene Expression Profiling
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Anti-Inflammatory Agents
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Neuronal Plasticity/physiology*
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Infarction/metabolism*
3.Influencing factors and predictive model construction of recurrence within one year after discontinuation of medication in children with cough variant asthma after regular treatment
Ye QIU ; Xiaoqian YANG ; Yachuan CAO ; Ying CUI
Journal of Clinical Medicine in Practice 2023;27(24):42-47
Objective To analyze the influencing factors of recurrence within one year after dis-continuation of regular treatment of children's cough variant asthma(CVA)and to construct a predic-tive model,in order to provide guidance for doctors to evaluate the risk of recurrence after CVA treat-ment in children and develop prevention strategies.Methods A total of 215 children with CVA were selected as the study objects.According to whether recurrence occurred within one year after discontin-uation of regular treatment,the patients were divided into recurrence group(n=89)and non-recur-rence group(n=126).Data of two groups were collected for univariate and multivariate Logistic re-gression analysis to screen out the influencing factors of recurrence in children with CVA.The GBM prediction model was constructed by using R software to run the gradient boosting model(GBM)algo-rithm.Receiver operating characteristic(ROC)curve was used to analyze and compare the predictive efficiency of GBM model and Logistic regression model.Results The recurrence rate of 215 children with CVA within one year after regular treatment was 41.40%(89/215).Allergy history(OR=2.870,95%CI,1.442 to 5.714),repeated respiratory infection history(OR=4.132,95%CI,2.045 to 8.348),parents'cognitive level of CVA prevention and control(OR=1.063,95%CI,1.011 to 1.118),eosinophils(OR=0.801,95%CI,0.674 to 0.952),forced expiratory flow at 50%of vi-tal capacity(FEF50)(OR=1.055,95%CI,1.012 to 1.099),maximal midexpiratory flow rate 75/25(MMEF75/25)(OR=1.054,95%CI,1.009 to 1.102),and fractional exhaled nitric oxide(FeNO)(OR=0.883,95%CI,0.834 to 0.935)were influencing factors for recurrence within one year after regular treatment of children's CVA(P<0.05).According to ROC curve analysis,the area under the curve of GBM model was 0.872,which was larger than 0.827 of Logistic regression model.Conclusion GBM prediction model is established based on the influencing factors of recur-rence within one year after regular treatment of CVA in children,which can be used as a reference for clinical measures to prevent recurrence in children after treatment of CVA.
4.Influencing factors and predictive model construction of recurrence within one year after discontinuation of medication in children with cough variant asthma after regular treatment
Ye QIU ; Xiaoqian YANG ; Yachuan CAO ; Ying CUI
Journal of Clinical Medicine in Practice 2023;27(24):42-47
Objective To analyze the influencing factors of recurrence within one year after dis-continuation of regular treatment of children's cough variant asthma(CVA)and to construct a predic-tive model,in order to provide guidance for doctors to evaluate the risk of recurrence after CVA treat-ment in children and develop prevention strategies.Methods A total of 215 children with CVA were selected as the study objects.According to whether recurrence occurred within one year after discontin-uation of regular treatment,the patients were divided into recurrence group(n=89)and non-recur-rence group(n=126).Data of two groups were collected for univariate and multivariate Logistic re-gression analysis to screen out the influencing factors of recurrence in children with CVA.The GBM prediction model was constructed by using R software to run the gradient boosting model(GBM)algo-rithm.Receiver operating characteristic(ROC)curve was used to analyze and compare the predictive efficiency of GBM model and Logistic regression model.Results The recurrence rate of 215 children with CVA within one year after regular treatment was 41.40%(89/215).Allergy history(OR=2.870,95%CI,1.442 to 5.714),repeated respiratory infection history(OR=4.132,95%CI,2.045 to 8.348),parents'cognitive level of CVA prevention and control(OR=1.063,95%CI,1.011 to 1.118),eosinophils(OR=0.801,95%CI,0.674 to 0.952),forced expiratory flow at 50%of vi-tal capacity(FEF50)(OR=1.055,95%CI,1.012 to 1.099),maximal midexpiratory flow rate 75/25(MMEF75/25)(OR=1.054,95%CI,1.009 to 1.102),and fractional exhaled nitric oxide(FeNO)(OR=0.883,95%CI,0.834 to 0.935)were influencing factors for recurrence within one year after regular treatment of children's CVA(P<0.05).According to ROC curve analysis,the area under the curve of GBM model was 0.872,which was larger than 0.827 of Logistic regression model.Conclusion GBM prediction model is established based on the influencing factors of recur-rence within one year after regular treatment of CVA in children,which can be used as a reference for clinical measures to prevent recurrence in children after treatment of CVA.
5.Specialty system of special physical education at master's level based on World Health Organization rehabilitation competency framework
Kai HUANG ; Yan LU ; Zhouying QIU ; Xiaoqian JIANG ; Chuanping HAO ; Zhe LI
Chinese Journal of Rehabilitation Theory and Practice 2022;28(3):318-326
Objective To explore the specialty system of special physical education at master's level.Methods Professional competency for special physical education was developed using the World Health Organization Rehabilitation Competency Framework (RCF), and was applied to the exploration of theoretical structure and methodological system for the construction of special physical education program at master's level.Results Based on the RCF, a competency framework for special physical education teachers was constructed, involving in five areas: practice, professionalism, learning and development, management and leadership, and research. The proficiency of competencies and activities should attain the corresponding level. The construction of special physical education program should be built to highlight the characteristics of RCF and focus on the training of competency in four areas: education objectives, instructional methods, the curriculum of the program and instructional assessment.Conclusion A competency-based program at the master's level in special physical education has been constructed using the RCF. The program uses the RCF to specify professional competency standards for special physical education teachers, and this can be transformed into the content of the competency-based program for special physical education at the master's level. The RCF is used to construct a competency-based special physical education system, which can be used to specify the educational objectives, instructional modalities, specialized core curriculum, and content and methods of instructional assessment in the design of a special physical education program at the master's level.
6.Construction and expression of the eukaryotic expression vector of wild-type p27 and lacking the nuclear localization signal p27
Xinyan JIAO ; Ying ZHANG ; Qiu SHENG ; Miao ZHANG ; Zejian YANG ; Xiaoqian GAO ; Qian ZHAO ; Bo WANG ; Peijun LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2022;43(4):516-521
【Objective】 To construct the eukaryotic expression vector carrying the human wild-type p27 and lacking nuclear localization signal p27△NLS coding sequences, and the express them in HEK293T cells, which may contribute to investigating the different locations and roles of p27 in the cytoplasm and nucleus. 【Methods】 Total RNA was prepared from human breast cancer MCF7 cells, and cDNA was obtained by reverse transcription-polymerase chain reaction (RT-PCR). After amplification of the p27 CDs and non-NLS fragments by PCR, full length p27WT (CDKN1B, NM_004064.5) and p27△NLS coding regions were obtained. PCR products were then subcloned into the eukaryotic expression vector pCMV-Blank. After identification with bacterial PCR, double restriction enzyme digestion and sequencing, they were defined officially as pCMV-p27WT and pCMV-p27△NLS, respectively. The recombinant plasmids were transfected into HEK293T cells by electroporation. After 48 h, the levels of p27 protein in the cytoplasm and nucleus were detected by Western blotting. 【Results】 The sequencing results showed that the sequences of p27WT and p27△NLS inserted into the plasmids were both correctly consistent with that of NM_004064.5. After transfection with pCMV-p27WT, total p27 protein expression was increased and distributed in both the cytoplasm and nucleus of HEK293T cells. After transfection with pCMV-p27△NLS, p27 protein was significantly increased and almost entirely localized in the cytoplasm of HEK293T cells. 【Conclusion】 The eukaryotic expression plasmids of human p27WT and p27△NLS coding sequences were successfully constructed and overexpressed in HEK293T cells. This research may lay a foundation for investigating the biological function of p27 in the cell cycle progression of tumor cells.
7.Does matching relation exist between the length and the tilting angle of terminal implants in the all-on-four protocol? stress distributions by 3D finite element analysis.
Xiaomei LI ; Zhizhong CAO ; Xiaoqian QIU ; Zhen TANG ; Lulu GONG ; Dalin WANG
The Journal of Advanced Prosthodontics 2015;7(3):240-248
PURPOSE: To explore whether there is matching relation between the length and the tilting angle of terminal implants in the All-on-Four protocol by studying the effects of different implant configurations on stress distributions of implant, bone, and framework. MATERIALS AND METHODS: Four implants were employed to support a full-arch fixed prosthesis and five three-dimensional finite element models were established with CT images, based on the length (S and L) and distal tilt angle (0degrees, 30degrees and 45degrees) of terminal implants for an edentulous mandible, which named: Tilt0-S, Tilt30-S, Tilt30-L, Tilt45-S and Tilt45-L. An oblique 240 N was loaded at second molar. The von Mises Stresses were analyzed. The implants were consecutively named #1 to #4 from the loading point. RESULTS: 1) Tilt0-S had the greatest stress on the implants, with the other groups exhibiting variable reductions; the four implants of Tilt45-L demonstrated the greatest reduction in stress. 2) Tilt0-S had the greatest stress at bone around #1 implant neck, and Tilt45-L exhibited the least stress, which was a 36.3% reduction compared to Tilt0-S. 3) The greatest stress in the framework was found on the cantilevers distal to #1 implant. Tilt45-S exhibited the least stress. CONCLUSION: Matching different length and tilting angle of the terminal implants led to variable stress reductions on implants, bone and the superstructure. By optimizing implant configuration, the reduction of stress on implants and surrounding bone could be maximized. Under the present condition, Tilt45-L was the preferred configuration. Further clinical testings are required.
Finite Element Analysis*
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Mandible
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Molar
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Neck
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Prostheses and Implants
8.Analysis of Bortezomib-containing combinations in newly-diagnosed multiple myeloma patients: a comparative study.
Lili XU ; Huiying QIU ; Xiaoxia HU ; Shuqing LYU ; Xianmin SONG ; Li CHEN ; Weiping ZHANG ; Lei GAO ; Xiaoqian XU ; Jianmin WANG ; Jianmin YANG
Chinese Journal of Hematology 2014;35(5):448-450
9.Mandibular first molar implant finite element models established by reverse engineering method
Qiang ZHU ; Dalin WANG ; Xiaoqian QIU ; Shaohai WANG
Chinese Journal of Tissue Engineering Research 2010;14(4):637-640
BACKGROUND: Reverse engineering modeling has been widely used in previous studies. However, there are a few studies on implantation modeling, especially in aspects of mandibular molar implantation. The mandibular molar has high miss rate in dentition. Thus, to establish three-dimensional finite element models of a jaw bones with mandibular molar implant denture using reverse engineering shows a great parameter value for research of mandibular molar implantation repair. OBJECTIVE: To establish a three-dimensional finite element model of a mandibular molar with dental implant inserted in mandibular molar area using reverse engineering, and to provide mathematical model basis for biomechanics researches of stomatology.METHODS: Establish three-dimensional model, divide mesh and calculate by Solidworks 2007 through scanning mandible by screw jaw using helix CT, obtaining data from images, separating data, rebuilding three-dimension using Mimics software,optimizing the design of mesh by Magics software, and dealing with solid model by Pro/E preprocessor.RESULTS AND CONCLUSION: A three-dimensional finite element model of mandibular first molar implant with regular depth was established, which has high geometrical similarity, good properties of mechanics and mesh, and can be easily modified. This can provide excellent model support for studying the stress analysis of different designs on restoring mandibular first molar implant.
10.The effect of the pillar implant system in the management of snoring and OSAHS patients.
Shaohua CHEN ; Siyi ZHANG ; Qianhui QIU ; Xiaoli SHENG ; Xiaoqian WANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2008;22(12):539-541
OBJECTIVE:
To explore the effect of the pillar implant system in the management of snoring and OSAHS patients.
METHOD:
All patients are divided into three groups by PSG: mild to moderate OSAHS, severe OSAHS and snoring. PSG and snoring visual analogue scale (VAS) were assessed preoperatively and postoperatively, and the postoperative PSG was performed after 6 months.
RESULT:
The values of VAS in all of the patients were reduced. AHI was reduced from 17.03 +/- 10.63 preoperatively to 11.17 +/- 9.52 postoperatively in 28 patients with mild to moderate OSAHS and the total effective ratio was 67.9%. AHI was reduced from 62.93 +/- 9.21 preoperatively to 55.42 +/- 13.77 postoperatively in 6 patients with severe OSAHS and the effective ratio was 16.7%.
CONCLUSION
The pillar implant system had a significant effect on mild to moderate OSAHS patients.
Adult
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Aged
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Female
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Humans
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Male
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Middle Aged
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Prostheses and Implants
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Prosthesis Implantation
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Sleep Apnea, Obstructive
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surgery
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Snoring
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surgery


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