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.Recent advances in the study of metabolic changes induced by arsenic exposure
Yifei SHAO ; Yuanyuan XIE ; Kun QIU ; Yiting CAI ; Fayu LU ; Shuhua XI
Chinese Journal of Endemiology 2024;43(6):513-516
Arsenic is a widely occurring metalloid element in the natural environment and is one of the primary carcinogens identified by the World Health Organization (WHO), but the specific carcinogenic mechanism is currently unclear. In recent years, through toxicological studies on arsenic, it has been found that exposure to arsenic can affect cellular metabolism in the body, which may be closely related to the carcinogenic mechanism of arsenic. Therefore, the authors review the research progress on arsenic exposure-induced effects on glucose metabolism, lipid metabolism, and amino acid metabolism, with a view to providing a theoretical basis for the study of the mechanism of arsenic carcinogenesis.
3.The role of NLRP3 signaling pathway in allergic rhinoconjunctivitis
Yubo GONG ; Xiaohua GUO ; Wen-Jun LU ; Yuanchao LI ; Changyu QIU ; Yuanyuan SHI ; Liping XIA ; Lin SHI ; Wei WU ; Ling LUO
The Journal of Practical Medicine 2024;40(14):1922-1927
Objective The objective of this study was to establish a mouse model of allergic rhinoconjunctivitis and investigate the role of the NLRP3 signaling pathway in allergic rhinoconjunctivitis.Methods Thirty-three female C57 mice(SPF)were randomLy divided into 3 groups:the control group,the experimental group,and the NLRP3-/-group.On days 0,4,7,14,and 21,the experimental group and NLRP3-/-group received a 0.2 mL intraperitoneal injection of medicine containing OVA(100 μg)and adjuvant Al(OH)3(4 mg),respectively.After an interval of 3 days,each eye and nose were dosed with 10 μL of 5%OVA for five consecutive days a week to induce allergic symptoms.During sensitization and excitation stages,the control group was replaced with an equiva-lent amount of PBS.Ocular and nasal symptoms were observed and scored.The levels of OVA-specific IgE,IL-4,IL-17,and IL-18 in serum were measured using ELISA,while changes in palpebral conjunctiva and nasal mucosa were assessed by hematoxylin-eosin staining.The expression of NLRP3 mRNA in conjunctival tissue and nasal mucosa was determined using real-time PCR analysis.Statistical analysis was performed using SPSS17.0 software with P<0.05 considered as statistically significant difference.Results The experimental group and NLRP3-/-group exhibited induced nasal and ocular allergic symptoms.In the experimental group,the duration of nasal allergy symptoms was(10.500±1.080)days,while the duration of eye allergy symptoms was(20.300±2.058)days.In the NLRP3-/-group,the duration of nasal allergy symptoms was(13.400±1.955)days,and for eye allergy symp-toms it was(20.900±2.132)days.The duration of nasal allergies in the NLRP3-/-group significantly exceeded that in the experimental group(P<0.05),whereas there were no significant differences observed in eye allergy durations between these two groups(P>0.05).Levels of OVA-specific IgE,IL-4,and IL-17 were significantly higher in both the experimental and NLRP3-/-groups compared to those in the control group(P<0.05).Additionally,serum IL-18 content increased significantly in the experimental group when compared with both control and NLRP3-/-groups(P<0.05).Conjunctival tissue lesions as well as nasal mucosa damage were evident in both experimental and NLRP3-/-groups.mRNA expression levels of NLRP3 within conjunctival tissue and nasal mucosa from the experimental group showed a significant increase when compared to those from both control and NLRP3-/-groups(P<0.05).Conclusion Allergic rhinoconjunctivitis pathogenesis is influenced by various factors;however,the involvement of NLPR3 signaling pathway promotes its development.
4.Infiltration and immunosuppressive function of tumor-associated B cells in gastric cancer patients
Yuxian LI ; Zhenquan DUAN ; Ying WANG ; Xueling TAN ; Xiaohong YU ; Yuanyuan ZHANG ; Baohang ZHU ; Yuan QIU ; Liusheng PENG ; Quanming ZOU
Journal of Army Medical University 2024;46(9):1034-1040
Objective To investigate the distribution of B cells in both tumor and non-tumor tissues of gastric cancer patients,analyze their phenotypic characteristics and explore the impact on T cell proliferation.Methods Immunohistochemical staining was utilized to detect the expression of B cell surface marker CD 19 in tumor and non-tumor tissues from 33 gastric cancer patients.The expression levels of chemokine receptors and immunoglobulin molecules on B cells in both tumor and non-tumor tissues were measured using flow cytometry.Chemotaxis experiments were conducted to examine the role of the CXCL12-CXCR4 axis in B cell chemotaxis.B cells isolated and purified from both tissue types were co-cultured with autologous peripheral T cells to assess their effect on T cell proliferation.Results There were significantly more B cells infiltrated in tumor tissues than those infitrated in the non-tumor tissues of gastric cancer patients(P<0.01),and CXCR4 was highly expressed on tumor-infiltrating B cells compared with B cells derived from non-tumor tissues(P<0.05).The Cancer Genome Atlas(TCGA)analysis indicated that the expression level of CXCL12 in tumor tissues was positively correlated with the expression level of CD19 in gastric cancer patients(r=0.15,P<0.01).And the expression level of CXCL12 in tumor tissues of the gastric cancer patients was also positively correlated with the number of B cells infiltrated in tumor tissues.Chemotaxis experiments confirmed that the CXCL12-CXCR4 axis was involved in promoting B cell chemotaxis(P<0.05).Although B cells in tumor and non-tumor tissues had similar levels of IgM,IgG,and IgA expression,tumor-infiltrating B cells significantly inhibited the proliferation of T cells when compared with B cells derived from non-tumor tissues(P<0.01).Conclusion There are more B cells infiltrated in gastric cancer tissues,which may be recruited to tumor tissues through the CXCL12-CXCR4 axis,and then inhibit T cell proliferation to promote the progression of gastric cancer.
5.Continuous vital signs monitoring using wireless wearable devices in patients after video-assisted thoracoscopic surgery for lung cancer: A prospective self-control study
Xiaoli MEI ; Yuchen HUANG ; Jian ZHOU ; Yuanyuan SONG ; Ailin LUO ; Mei YANG ; E ZHENG ; Yang QIU ; Beinuo WANG ; Zhenghao DONG ; Hu LIAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(02):229-235
Objective To explore the reliability and safety of continuous monitoring of vital signs in patients using wireless wearable monitoring devices after video-assisted thoracoscopic surgery (VATS) for lung cancer. Methods The patients undergoing VATS for lung cancer in West China Hospital, Sichuan University from May to August 2023 were prospectively enrolled. Both wireless wearable and traditional wired devices were used to monitor the vital signs of patients after surgery. Spearman correlation analysis, paired sample t test and ratio Bland-Altman method were used to test the correlation, difference and consistency of monitoring data measured by the two devices. The effective monitoring rate of the wireless wearable device within 12 hours was calculated to test the reliability of its continuous monitoring. Results A total of 20 patients were enrolled, including 15 females and 5 males with an average age of 46.20±11.52 years. Data collected by the two monitoring devices were significantly correlated (P<0.001). Respiratory rate and blood oxygen saturation data collected by the two devices showed no statistical difference (P>0.05), while heart rate measured by wireless wearable device was slightly lower (=−0.307±1.073, P<0.001), and the blood pressure (=1.259±5.354, P<0.001) and body temperature(=0.115±0.231, P<0.001) were slightly higher. The mean ratios of heart rate, respiratory rate, blood oxygen saturation, blood pressure and body temperature collected by the two devices were 0.996, 1.004, 1.000, 1.014, and 1.003, respectively. The 95% limits of agreement (LoA) and 95% confidence interval of 95%LoA of each indicator were within the clinically acceptable limit. The effective monitoring rate of each vital signs within 12 hours was above 98%. Conclusion The wireless wearable device has a high accuracy and reliability for continuous monitoring vital signs of patients after VATS for lung cancer, which provides a security guarantee for subsequent large-scale clinical application and further research.
6.Effect and its mechanism of esketamine on anxiety and depression in mice
Jingwen Zhou ; Yuanhai Li ; Gaolin Qiu ; Wen Cai ; Yuanyuan Zhao ; Xiaoqiong Xia
Acta Universitatis Medicinalis Anhui 2024;59(1):106-110
Objective :
To explore the effect of esketamine on anxiety-depressive-like behavior in mice and its rela- tionship with inflammation.
Methods :
SPF grade healthy adult male C57BL/6J mice,weighing 20 -24 g,were used in the exprement.The random number table method was used to divide into 5 groups (n = 8) : control group ( Con group) ,esketamine group (ESK group) ,model group ( LPS group) ,model + esketamine prevention group (LPS + ESK1 group) and model + esketamine treatment group ( LPS + ESK2 group) .An inflammation-induced anxiety-depression model was prepared by intraperitoneal injection of LPS 0. 83 mg / kg.The ESK group was injec- ted with esketamine 10 mg / kg ; LPS group was injected with LPS 0. 83 mg / kg ; LPS + ESK1 group was injected with LPS 0. 83 mg / kg before 24 hours intraperitoneal injection of esketamine 10 mg / kg ; and the LPS + ESK2 group was injected with LPS 0. 83 mg / kg and 30 minutes later with esketamine 10 mg / kg.24 h after intraperitoneal injec- tion of LPS,the anxiety-depression-like behaviors of mice were measured using behavioral experiments.At the end of behavioral experiments,the spleen was taken immediately ; hippocampal tissues were taken and enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of interleukin-1 β (IL-1 β) ,tumor necrosis factor al- pha (TNF-α) and interleukin 6 (IL-6) and neuronal pathological changes in hippocampal tissues were observed by HE staining.
Results :
Compared with the Con group,mice in the LPS group showed increased anxiety and depres- sion-like behavior (P<0. 05) ,increased spleen weight / body weight (P <0. 05 ) ,increased hippocampal tissue concentrations of IL-1 β , TNF-α and IL-6 (P<0. 05) ,and increased neuronal degeneration necrosis,there was no statistically significant difference in these indicators in the ESK group compared with the Con group.Compared with the LPS group,mice in the LPS + ESK1 and LPS + ESK2 groups showed reduced anxiety-depression-like behavior (P<0. 05) ,decreased splenic weight / body weight (P <0. 05) ,hippocampal tissue IL-1 β , TNF-α , IL- 6 con- centrations were reduced (P<0. 05) ,and neuronal degeneration necrosis was reduced.Compared with the LPS + ESK1 group,the LPS + ESK2 group showed an increase in the distance travelled in the central area of the open field experiment and the distance into the open arm of the elevated cross maze experiment (P<0. 05) ,a decrease in IL-6 and TNF-α concentrations (P<0. 05) ,and a reduction in the degree of neuronal damage.
Conclusion
Esketamine ameliorates LPS-induced anxiety-depression-like behavior and neuronal damage in mice by a mechanism that may be related to reduced inflammation.
7.Analysis on prevalence of chronic kidney disease in adults in Qingdao, 2019-2020
Longfei QIU ; Yuanyuan ZHAO ; Hua ZHANG ; Xiaojia XUE ; Yang ZHAO ; Yaoyao XU ; Haiping DUAN
Chinese Journal of Epidemiology 2024;45(5):694-699
Objective:To understand the prevalence of chronic kidney disease (CKD) in adults in Qingdao.Methods:A multi-stage stratified random sampling method was used to select 6 240 local residents aged ≥18 years in Qingdao as study subjects from May 2019 to September 2020, the demographic data of the study subjects were collected by face-to-face survey method. The prevalence of CKD in adults in Qingdao were analyzed using software SPSS 25.0.Results:A total of 5 996 adults in Qingdao were included in this study. The overall prevalence rate of CKD in the adults was 8.22%. The prevalence rates of CKD in men and women were 7.70% and 8.74%, respectively. The prevalence rate of CKD was 10.28% in urban residents and 7.25% in rural residents, the differences in the prevalence of CKD among different age, educational level and marital status groups were significant ( P<0.001). The prevalence of CKD tended to increase with age and decrease with the increase of education level. Conclusions:The prevalence of CKD in adults of Qingdao was relatively high. It is necessary to actively carry out the early prevention and treatment of CKD and strengthen the screening and prevention of CKD to reduce the incidence and slow development of CKD.
8.Application of remote insulin injection service in home management of patients with type 2 diabetes mellitus
Jinwei QIU ; Xiaoling QU ; Yuanyuan LI
Chinese Journal of Modern Nursing 2024;30(1):101-105
Objective:To explore the application effect of remote insulin injection service in home management of type 2 diabetes mellitus.Methods:A total of 134 patients with type 2 diabetes who were hospitalized in Department of Endocrinology in Beijing Friendship Hospital Affiliated to Capital Medical University from March to December 2022 were selected as research objects by the convenient sampling method. According to the random number table method, the patients were divided into the control group and the observation group, with 67 cases in each group. After discharge, the control group received routine out-of-hospital home follow-up management, and the observation group received remote insulin injection service management on the basis of the control group. The management period was three months. Differences in glycemic indicators, Scale of the Diabetes Self-Care Activities (SDSCA) score, insulin injection compliance and incidence of insulin injection-related negative events were compared between the two groups.Results:A total of 134 questionnaires were sent out in this study, and 134 were effectively collected, with an effective recovery rate of 100.00%. After management, fasting blood glucose and glycosylated hemoglobin in the observation group were lower than those in the control group, and SDSCA score was higher than that in the control group, and the differences were statistically significant ( P<0.01). Moreover, fasting blood glucose and glycosylated hemoglobin in observation group after management were lower than before management, and SDSCA score was higher than before management, and the differences were statistically significant ( P<0.05). The insulin injection compliance of the observation group was better than that of the control group, and the difference was statistically significant ( Z=2.485, P=0.013). The total incidence of negative events in the observation group were lower than those in the control group, and the differences were statistically significant ( P<0.01) . Conclusions:Remote insulin injection service can effectively improve the blood glucose control level of patients with type 2 diabetes mellitus at home, improve self-management ability and insulin injection compliance and reduce the incidence of negative events related to insulin injection, which is worthy of clinical promotion and application.
9.Construction of recombinant CVI988 vector vaccine integrating IBDV-VP2 gene
Lele GONG ; Xinxiang HUANG ; Yunzhe KANG ; Lele WANG ; Xiangqi QIU ; Yuanyuan ZHANG ; Meijie GAO ; Wenhui ZHU ; Yulin ZHANG ; Guoqing ZHUANG ; Aijun SUN
Chinese Journal of Veterinary Science 2024;44(9):1865-1871
The emergence of high virulent mutant strains of infectious bursal disease virus(IBDV)becomes a serious threat to the poultry industry.However,the live attenuated IBDV vaccine can potentially revert to a virulent strain.Therefore,it is a necessary to develop safe and effective IB-DV-associated vaccines.The construction of a recombinant Marek's disease(MD)vaccine strain,CVI988,expressing the IBDV VP2 protein,can protect against disease induced by both IBDV and Marek's disease virus(MDV).Here,the IBDV-VP2 gene was integrated into the UL55 locus of CVI988 by bacterial artificial chromosome(BAC)technique,resulting in the recombinant virus CVI988 BAC-VP2.The recombinant virus was characterized by PCR,IFA and subsequently the bi-ological properties of the recombinant virus were investigated.The results showed that the recom-binant virus CVI988 BAC-VP2 was successfully rescued.The VP2 protein stably expressed in chick-en embryo fibroblasts(CEF).The growth kinetics and plague size assays showed that there was comparable replication ability between recombinant virus and parental virus.This study provides the basis for the development of a low-cost vaccine against both IBDV and MDV infections.
10.Advances in safety studies of gluteal fat grafting
Julei ZHANG ; Xiaodong LI ; Zeliang HE ; Chengliang ZHANG ; Yuanyuan YAO ; Lingling LIU ; Liang’en AN ; Jin LI ; Shulin QIU
Chinese Journal of Plastic Surgery 2023;39(5):561-564
The gluteal fat grafting technique continues to grow in popularity, along with which the safety issue should not be ignored. It drew attentions of researchers that due to the existence of large vessels, fat embolism or fat embolism syndrome occurred more frequently for gluteal fat grafting, which might lead to death. In this situation, researchers performed a series of study including retrospective case series, systematic analysis, questionnaire survey and anatomical study, and summarized a series of safety principles for gluteal fat grafting, such as scanning high-risk patients; staying subcutaneously injection, avoiding intramuscular injection; using cannula size over 4 mm; keeping cannula parallel to gluteus maximus; injecting while withdrawing; and avoiding excessive local pressure. Based on the literature review, this article analyzes and summarizes the safety of gluteal fat grafting.


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