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1.
Chinese Journal of Schistosomiasis Control ; (6): 1-6, 2024.
Article in Chinese | WPRIM | ID: wpr-1013562

ABSTRACT

The goal of achieving elimination of schistosomiasis across all endemic counties in China by 2030 was proposed in the Outline of the Healthy China 2030 Plan. On June 16, 2023, the Action Plan to Accelerate the Elimination of Schistosomiasis in China (2023—2030) was jointly issued by National Disease Control and Prevention Administration and other 10 ministries, which deployed the targets and key tasks of the national schistosomiasis elimination programme in China. This article describes the progress of the national schistosomiasis control programme, analyzes the opportunities to eliminate schistosomiasis, and proposes targeted recommendations to tackle the challenges of schistosomiasis elimination, so as to accelerate the process towards schistosomiasis elimination and facilitate the building of a healthy China.

2.
Shanghai Journal of Preventive Medicine ; (12): 619-625, 2023.
Article in Chinese | WPRIM | ID: wpr-988894

ABSTRACT

ObjectiveTo analyze the epidemic trends and current status in the prevalence of schistosomiasis in Jiangxi Province from 2002 to 2021, and to provide evidence for formulating and optimizing schistosomiasis control strategies in the province. MethodsEpidemiological data of schistosomiasis in Jiangxi Province from 2002 to 2021 were collected. The Joinpoint regression model was used to investigate the epidemic trends of schistosomiasis, and the annual average percent change (AAPC) was used to assess the changing trend of schistosomiasis infection in humans and bovines. SPSS 25.0 software was used to analyze the target achievement status, infection rate of schistosomiasis in humans and the areas with snails in 2021 to evaluate the differences among various districts and epidemic regions. ResultsBy 2021, 24 counties (61.50%) in the province had achieved the elimination goal, with hilly and marsh counties reaching 82.60% and 31.30%, respectively. The seropositive rate of schistosomiasis in human was 3.51% across the province. However, the seropositive rate in marshland areas (4.77%) was significantly higher than that in hilly areas (1.23 %) (χ2=3 827.51, P<0.001). The total area with snails in the province was 84 938.32 hm2, including 82 196.86 hm2 in marshland areas and 2 741.46 hm2 in hilly areas. From 2002 to 2021, the infection rate of schistosomiasis in the humans and bovines exhibited a declining trend, with AAPC values of -48.70 and -39.70, respectively. The decline rate in the hilly areas was higher than that in marshland areas. However, the area with snail showed an upward trend (AAPC=0.50), with a growth rate of 5.80% in hilly areas, which was greater than that in marshland areas (0.40%). ConclusionFrom 2002 to 2021, the infection rate of schistosomiasis in humans and bovines in Jiangxi Province continued to decline, while the areas with snails increased. The prevention focus should still be on the marshland endemic areas, and attention should also be given to the risk of schistosomiasis in hilly endemic areas. Therefore, it is necessary to continue implementing a comprehensive prevention and control strategy that emphasizes controlling the sources of infection and adopts tailored measures based on local conditions and precise policies.

3.
Chinese Journal of Schistosomiasis Control ; (6): 641-645, 2023.
Article in Chinese | WPRIM | ID: wpr-1011424

ABSTRACT

Schistosomiasis control is not only a disease control programme, but also a great social practice activity in China. During the evolution of national schistosomiasis control programmes, the special schistosomiasis control culture has been cultivated and developed, which contains the spiritual connotation of government-led, people-oriented, respect for science and integration of all efforts. The publication of Chairman Mao Zedong’s two poems entitled “Farewell to the God of Plague” and the post-script in 1958 was a sign for the formation and development of Chinese schistosomiasis control culture, which always lead the orientation of development and practice of schistosomiasis control culture building. The schistosomiasis control culture provides powerful spiritual motivation and supports to schistosomiasis control programmes in China, and improving the building of schistosomiasis control culture is of great significance to strengthen our belief in achieving the goal of schistosomiasis elimination, mobilize all social resources, accelerate the progress towards elimination of schistosomiasis and facilitate the high-quality development of healthcare services. Chinese schistosomiasis control spirit is the refinement from the cultural connotation of the long-term schistosomiasis control programmes in China, and is the most essential and concentrated embodiment of the schistosomiasis control culture. This article describes the great significance of two poems entitled “Farewell to the God of Plague”, summarizes the connotation and role of schistosomiasis control spirit, and introduces the practice, development and innovation of schistosomiasis control culture building in Jiangxi Province.

4.
Chinese Journal of Applied Clinical Pediatrics ; (24): 1903-1906, 2021.
Article in Chinese | WPRIM | ID: wpr-930354

ABSTRACT

Respiratory viruses can cause a variety of serious respiratory infections and diseases of tissues and organs outside the respiratory tract, raising a potentially severe threat to the society.Virus replication and survival rely on the internal mechanism of host cells, and the latter also produce a variety of restriction factors that target viral invasion, genome transcription and replication, and assembly and release to block viral infection.Herein, this study reviewed the research progress of the antiviral effects of the host restriction factors of common respiratory viruses and their underlying mechanisms.

5.
Chinese Journal of Radiology ; (12): 124-128, 2020.
Article in Chinese | WPRIM | ID: wpr-799429

ABSTRACT

Objective@#To analyze the function of essential hypertension′s working memory network by using task-fMRI and independent component analysis (ICA).@*Methods@#According to the enrollment criteria, 24 hypertensive patients and 23 healthy controls (May 2016 to January 2017) in the 900 Hospital of the Joint Service Support Force were collected, who underwent task-state functional MRI data acquisition of N-back by using Siemens 3.0 T MRI. All subjects completed neuropsychological cognitive tests, blood pressure measurements and biochemical tests before MRI. Task-state functional MRI was analyzed by SPM 12, and independent component analysis was performed by GIFT 4.0 software. The independent-samples t-test was used to analyze the maps of the two groups′s task-state fMRI and ICA. The results were adjusted by multiple comparison correction of Alphasim, P<0.05 showed that the difference was statistically significant.@*Results@#Compared with the healthy group, the activation of left superior frontal gyrus and left middle frontal gyrus in hypertension group was enhanced (P<0.05), but there was no obvious weakened activation area. In the independent component analysis, the functional connectivity of left middle frontal gyrus and left anterior central gyrus in hypertension group was significantly lower than that in healthy control group (P<0.05), and there was no significant enhancement of functional connectivity.@*Conclusions@#The working memory network of patients with essential hypertension may change abnormally due to the influence of long-term hypertension. This kind of damage can compensate through the functional enhancement of brain regions, thus maintaining the normal cognitive level of hypertensive patients.

6.
Chinese Journal of Radiology ; (12): 124-128, 2020.
Article in Chinese | WPRIM | ID: wpr-868261

ABSTRACT

Objective:To analyze the function of essential hypertension′s working memory network by using task-fMRI and independent component analysis (ICA).Methods:According to the enrollment criteria, 24 hypertensive patients and 23 healthy controls (May 2016 to January 2017) in the 900 Hospital of the Joint Service Support Force were collected, who underwent task-state functional MRI data acquisition of N-back by using Siemens 3.0 T MRI. All subjects completed neuropsychological cognitive tests, blood pressure measurements and biochemical tests before MRI. Task-state functional MRI was analyzed by SPM 12, and independent component analysis was performed by GIFT 4.0 software. The independent-samples t-test was used to analyze the maps of the two groups′s task-state fMRI and ICA. The results were adjusted by multiple comparison correction of Alphasim, P<0.05 showed that the difference was statistically significant. Results:Compared with the healthy group, the activation of left superior frontal gyrus and left middle frontal gyrus in hypertension group was enhanced ( P<0.05), but there was no obvious weakened activation area. In the independent component analysis, the functional connectivity of left middle frontal gyrus and left anterior central gyrus in hypertension group was significantly lower than that in healthy control group ( P<0.05), and there was no significant enhancement of functional connectivity. Conclusions:The working memory network of patients with essential hypertension may change abnormally due to the influence of long-term hypertension. This kind of damage can compensate through the functional enhancement of brain regions, thus maintaining the normal cognitive level of hypertensive patients.

7.
Chinese Journal of Anesthesiology ; (12): 1181-1184, 2019.
Article in Chinese | WPRIM | ID: wpr-824684

ABSTRACT

Objective To evaluate the role of hippocampal CD200 receptor 1(CD200R1)in peri-operative neurocognitive disorders(PND)in mice.Methods Sixty clean-grade male C57BL/6 mice,aged 9-10 months,weighing 32-38 g,were used in the study.The experiment was performed in two parts.Ex-periment Ⅰ Thirty-six mice were divided into 2 groups(n=18 each)using a random number table meth-od: control group(group C)and PND group.Group C only received isoflurane anesthesia.Partial left lo-bectomy of the liver was performed under isoflurane anesthesia in group PND.Contextual fear conditioning test was performed at 1,3 and 7 days after surgery,and the freezing time was recorded.The mice were then sacrificed,and the hippocampus was isolated for determination of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)contents(by enzyme-linked immunosorbent assay)and CD200 and CD200R1 expression(by Western blot).ExperimentⅡ Twenty-four mice were divided into 2 groups(n=12 each)using a random number table method: CD200-Fc group and IgG1-Fc group.Recombinant proteins CD200-Fc and human IgG1-Fc were injected into the lateral cerebral ventricle in CD200-Fc group and IgG1-Fc group,respectively.Partial left lobectomy of the liver was performed after the end of injection.Contextual fear conditioning test was performed at 1 and 3 days after surgery,and the freezing time was recorded.Re-sults Experiment Ⅰ Compared with group C,the freezing time in the contextual fear conditioning test was significantly shortened,and the contents of IL-1β were increased at 1 and 3 days after surgery,the contents of TNF-α were increased at 3 and 7 days after surgery,and the expression of CD200 and CD200R1 was up-regulated at 1 day after surgery in group PND(P<0.05).ExperimentⅡ Compared with IgG1-Fc group,the freezing time in the contextual fear conditioning test was significantly prolonged at 1 day after surgery in CD200-Fc group(P<0.05).Conclusion Up-regulated expression of hippocampal CD200R1 is the endogenous protective mechanism of PND in mice.

8.
Chinese Journal of Anesthesiology ; (12): 1181-1184, 2019.
Article in Chinese | WPRIM | ID: wpr-797053

ABSTRACT

Objective@#To evaluate the role of hippocampal CD200 receptor 1 (CD200R1) in perioperative neurocognitive disorders (PND) in mice.@*Methods@#Sixty clean-grade male C57BL/6 mice, aged 9-10 months, weighing 32-38 g, were used in the study.The experiment was performed in two parts.Experiment Ⅰ Thirty-six mice were divided into 2 groups (n=18 each) using a random number table method: control group (group C) and PND group.Group C only received isoflurane anesthesia.Partial left lobectomy of the liver was performed under isoflurane anesthesia in group PND.Contextual fear conditioning test was performed at 1, 3 and 7 days after surgery, and the freezing time was recorded.The mice were then sacrificed, and the hippocampus was isolated for determination of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) contents (by enzyme-linked immunosorbent assay) and CD200 and CD200R1 expression (by Western blot). Experiment Ⅱ Twenty-four mice were divided into 2 groups (n=12 each) using a random number table method: CD200-Fc group and IgG1-Fc group.Recombinant proteins CD200-Fc and human IgG1-Fc were injected into the lateral cerebral ventricle in CD200-Fc group and IgG1-Fc group, respectively.Partial left lobectomy of the liver was performed after the end of injection.Contextual fear conditioning test was performed at 1 and 3 days after surgery, and the freezing time was recorded.@*Results@#Experiment Ⅰ Compared with group C, the freezing time in the contextual fear conditioning test was significantly shortened, and the contents of IL-1β were increased at 1 and 3 days after surgery, the contents of TNF-α were increased at 3 and 7 days after surgery, and the expression of CD200 and CD200R1 was up-regulated at 1 day after surgery in group PND (P<0.05). Experiment Ⅱ Compared with IgG1-Fc group, the freezing time in the contextual fear conditioning test was significantly prolonged at 1 day after surgery in CD200-Fc group (P<0.05).@*Conclusion@#Up-regulated expression of hippocampal CD200R1 is the endogenous protective mechanism of PND in mice.

9.
Chinese Journal of Schistosomiasis Control ; (6): 397-401, 2017.
Article in Chinese | WPRIM | ID: wpr-615677

ABSTRACT

China's prevention and control of parasitic diseases has made remarkable achievements. However,the prevalence and transmission of parasitic diseases is impacted by the complicated natural and social factors of environment ,natural disas-ters,population movements,and so on. Therefore,there are still the risks of the outbreak of emergency parasitic diseases af-fairs,which may affect the control effectiveness of parasitic diseases and endanger the social stability seriously. In this article , we aim at the analysis of typical cases of emergency parasitic disease affairs and their impacts on public health security in China in recently years,and we also elaborate the disposal characteristics of emergency parasitic disease affairs,and propose the estab-lishment of response system to emergency parasitic disease affairs in China,including the organizational structure and response flow path,and in addition,point out that,in the future,we should strengthen the system construction and measures of the re-sponse system to emergency parasitic disease affairs,so as to control the risk and harm of parasitic disease spread as much as possible and to realize the early intervention and proper disposal of emergency parasitic disease affairs.

10.
Chinese Journal of Nuclear Medicine and Molecular Imaging ; (6): 538-543, 2017.
Article in Chinese | WPRIM | ID: wpr-607126

ABSTRACT

Objective To evaluate the diagnostic value of the SPM-based interhemispheric asymmetry analysis (SIAA) of 18F-FDG PET brain imaging in localization of temporal lobe epileptic foci.Methods A total of 73 patients (42 males,31 females,average age: (27.10±9.78) years) diagnosed as temporal lobe epilepsy (TLE) and 30 healthy volunteers (17 males,13 females,average age: (33.79±5.46) years) from March 2012 to July 2015 were enrolled in this retrospective study.18F-FDG PET brain imaging was performed on all subjects to localize temporal lobe epileptic foci.The results of 18F-FDG-PET were analyzed by SIAA,visual analysis and asymmetry index (AI) analysis.The accuracy of the three methods was evaluated according to the postoperative pathological results and curative effect.χ2 test and partition of χ2 test were used to analyze the data.Results The accuracies of localization with SIAA,visual analysis and AI analysis were 87.67%(64/73),75.34%(55/73) and 69.86%(51/73),respectively (χ2=6.993,P0.017).Conclusion The SIAA of 18F-FDG PET brain imaging may be a more objective and accurate method in localization of TLE foci.

11.
The Journal of Practical Medicine ; (24): 1074-1077, 2017.
Article in Chinese | WPRIM | ID: wpr-619075

ABSTRACT

Objective To evaluate the effect of silencing ACAT1 gene on colon cancer cells proliferation,migration,invasion and colon cancer development by using the small interference RNA (siRNA) in colon cancer cell line HT-29.Methods Acyl coenzyme A cholesterol acyltransferase 1 (ACAT1) gene was silenced in HT-29 cell lines using Hiperfect transfection reagent.The expression level of ACAT1 was detected by real time PCR.CFSE and transwell assays were used to evaluate the effect of ACAT1 gene interfering on cells proliferation,mi gration and invasion.Result ACAT1 mRNA expression decreased obviously after siRNA interference.Compared with pre-transfection,proliferation,migration and invasion of colon cancer cells have been significantly inhibited (P < 0.05).Conclusion ACAT1 gene interference reduced proliferation,migration and of invasion of HT29 cells,which provide a new potential target for colon cancer treatment.

12.
Chinese Journal of Schistosomiasis Control ; (6): 644-647,682, 2016.
Article in Chinese | WPRIM | ID: wpr-605925

ABSTRACT

Objective To explore the cut?off value of the indirect haemagglutination test(IHA)method for schistosomiasis japonica diagnosis in different endemic areas. Methods Totally 55 nature villages of the lake?type endemic counties,Yugan and Xinzi,in Poyang Lake Region of Jiangxi Province were chosen as the study fields,and all the villagers over 5 years old were parallelly examined by Kato?Katz method+miracidial hatching test and IHA method. The detection data were analyzed by the correlation analysis,and the threshold values of the IHA method in different endemic areas were decided by the receiver operat?ing characteristic(ROC)curve. Results The positive rate of stool examinations of the villagers was correlated with the distribu?tion trend of the antibody level of whole population(r=0.588,P0.05). The antibody level of stool?negative population during the period of 2008 to 2011 detect?ed by IHA method dropped year by year,and the annual difference was statistically significant(F=3.650,P0.05). When the positive rates were5%,the specificity of diagnosis could be improved when 1∶80,1∶20 and 1∶10 were used as the cut?off val? ues of IHA correspondingly. Conclusion The different threshold values for diagnosis of schistosomiasis japonica should be con?sidered while using IHA method to screen out patients in different endemic areas.

13.
Chinese Journal of Radiology ; (12): 576-580, 2016.
Article in Chinese | WPRIM | ID: wpr-502017

ABSTRACT

Objective To explore early aberrant change of the function and grey matter volume within default-mode network in patients with type 2 diabetes mellitus (T2DM) by Independent component analysis and voxel-based morphometry.Methods Twenty-nine T2DM patients and thirty-three healthy control subjects were recruited in this study between May 2014 and January 2015,and the demographic and clinical data (fasting plasma glucose and HbA1c level) were obtained.General cognition was assessed by MMSE and MoCA in all subjects.Resting-state fMRI and T1-weighted imaging were performed in all subjects.Then the z-maps of default-mode network were obtained from Resting-state fMRI by independent component analysis,and the grey matter volumes were extracted by VBM.Finally,two-sample t-test was performed in the function and grey matter volume of DMN between two groups,and pearson correlation coefficients among zFC values extracted from significantly different brain regions,grey matter volumes and durations of T2DM patients were calculated.Results Patients with T2DM showed decreased Z value around the right anterior cingulated cortex and the right medial pre-frontal cortex(cluster=121,t=-3.81,P< 0.05).There was no significant difference in grey matter volume between the two groups,and no correlation between Z value or duration and grey matter volume was found.Conclusions It suggests the abnormality of connectivity of default-mode network may precede the abnormality of grey matter volume in type 2 diabetes mellitus patients.The DMN might be a new potential tool in monitoring the change of cognition in patients with type 2 diabetes mellitus.

14.
Chinese Journal of Schistosomiasis Control ; (6): 456-460, 2016.
Article in Chinese | WPRIM | ID: wpr-495737

ABSTRACT

This report analyzes the status of schistosomiasis epidemic,surveillance and challenges in transmission?interrupt?ed areas of Jiangxi Province. There are 24 schistosomiasis transmission interrupted counties(cities,districts)among 39 endem?ic counties(cities,districts). Among the 24 counties,there are 6 counties with Oncomelania hupensis snails and 461 villages endemic,with 1.29 million residents. Since 2009,there have not been acute and chronic cases of schistosomiasis and all the pa?tients are historical advanced schistosomiasis cases. The area with snail habitats is 37.02 hm2. Since 2015,the schistosomiasis surveillance has covered the entire transmission interrupted areas. However,there are still many challenges,such as endemic in?stability,weak surveillance capacity,lack of surveillance awareness and other issues. Towards the schistosomiasis elimination, the surveillance should be based on the snail monitoring mainly,supplemented by the mobile population monitoring,and in ad?dition,the capacity building,standardized report of endemic,and snail habitat reform should also be further strengthened.

15.
Chinese Journal of Schistosomiasis Control ; (6): 11-17, 2016.
Article in Chinese | WPRIM | ID: wpr-491851

ABSTRACT

Objective To explore the immune mechanism of negative results of immune tests of schistosomiasis japonica pa?tients. Methods Totally 142 schistosomiasis patients(positive stool examinations)of Poyang Lake region were tested by ELI?SA method,and the ROC curve was applied to determine the high and low response of the patients. The levels of cellular immu?nity and cytokines of high and low responders were compared. Results Totally eight schistosomiasis patients were found as low responders. Besides SWAP?IgA(t= -1.588,P > 0.1),the levels of isotype antibodies were significantly lower in the low re?sponders compared with those in the high responders(t = -14.517 to -2.866,all P 0.05)compared with those in the high responders. The differences of IFN?γ and IL?10 between the high and low responders were both not significant(t= -2.426 to 0.216,all P >0.05). Conclusions There is a significant difference between the high and low responders only in the levels of isotype antibod?ies. One of the reasons of low response in the immune tests is the much lower antibody level after the antigen?antibody compound is completely formulated.

16.
Chinese Journal of Schistosomiasis Control ; (6): 58-61, 2016.
Article in Chinese | WPRIM | ID: wpr-491799

ABSTRACT

Objective To design and develop new types of health education materials which are suitable for fishermen and boatmen in endemic areas of marshland and lake regions,and to observe their application effects. Methods A total of 292 adult fishmen and boatmen who lived in Houshan Village,Yugan County,the schistosomiasis endemic area of Poyang Lake Re?gion,were selected randomly and investigated by questionnaires to understand the status of their knowledge,attitudes,practic?es on schistosomiasis control as well as the channels for getting information on schistosomiasis control and the materials that they were willing to accept. Then the information and materials suitable for the target population were developed together by the re?searchers and the volunteers of the villagers through focus group discussions,personal interviews and the Delphi method. Re?sults A series of participatory health education materials of schistosomiasis control targeted to the fishmen and boatmen were developed,including 2 live posters,2 picture puzzles,2 short opusculums and one song about schistosomiasis control. The field application showed that 98.97%,84.38%,78.35% and 80.93% of the participants considered those materials had scientificity, intelligibility,interestingness and practicability,respectively. Conclusion The participatory health education materials of schistosomiasis control is suitable for fishmen and boatmen,which can be used for reference by other endemic areas in marsh?land and lake regions.

17.
Chinese Journal of Schistosomiasis Control ; (6): 561-564,574, 2014.
Article in Chinese | WPRIM | ID: wpr-600245

ABSTRACT

Schistosomiasis is closely related to natural environmental factors. The changes of environmental factors such as temperature hydrology vegetation soil etc. all impact the scope and extent of schistosomiasis transmission. Poyang Lake is the largest freshwater lake and one of the major endemic areas with schistosomiasis in China. With global warming the imple-mentation of the Three Gorges Dam operation and the Poyang Lake Ecological Economic Planning the natural environment in Poyang Lake area has been and will continue to change especially the water environment and climate environment which are more closely related to the schistosomiasis transmission. These changes to some extent have affected and will continue to affect the prevalence and transmission of schistosomiasis. This article reviews the relationship between the natural environment and its changes and schistosomiasis transmission in the Poyang Lake region.

18.
Chinese Journal of Schistosomiasis Control ; (6): 479-481, 2014.
Article in Chinese | WPRIM | ID: wpr-459684

ABSTRACT

In this study,we summarized the results from the retrospective investigation on endemic situation of schistosomia-sis that was implemented in nine provinces(autonomous region),China in 2009,demonstrated the role of these retrospective in-vestigations in accelerating the progress of schistosomiasis control in China,and clarified the great significance of the investiga-tion for summarizing the experiences for the control of schistosomiasis,and analyzing the changing patterns and affecting factors of endemic status of schistosomiasis in China. In addition,these retrospective investigations provide reliable evidence for revis-ing the Criteria of Schistosomiasis Control and Elimination,and for the more accurate and scientific assessment of the effec-tiveness of schistosomiasis control in China.

19.
Chinese Journal of Schistosomiasis Control ; (6): 250-253, 2014.
Article in Chinese | WPRIM | ID: wpr-451091

ABSTRACT

Objective To understand the characteristics of schistosome cercarial infested water contact of residents in Poyang Lake endemic areas after the operation of Three Gorges Project,so as to provide the evidence for formulating a well-directed con-trol strategy. Methods The residents in two villages in schistosomiasis endemic areas were selected and surveyed for their activi-ties of infested water contacts by questionnaire. Results The proportion and frequency of infested water contacts of residents in Poyang Lake region were high and the average rate of infested water contacts was 69.77%,and 98.96%of infested water contacts occurred in spring,summer and autumn. The main ways of infested water contacts were fishing/swimming/playing for adult men, washing clothes/fishing for adult women,and swimming/playing for school children,respectively. The rates of infested water con-tacts were 73.33%,71.63%and 61.87%in adult men,women and school children,respectively. The frequencies of infested water contacts>1 time/week,1 time/week-1 time/month,and<1 time/month were 62.61%,23.74%and 13.66%respectively. Conclu-sions The infested water contacts mainly occurs in spring,summer and autumn,and the main way of infested water contacts was fishing. Farming and mowing grass,which had been the primary way,now become the secondary way for the adult residents.

20.
Chinese Journal of Microbiology and Immunology ; (12): 112-116, 2013.
Article in Chinese | WPRIM | ID: wpr-436454

ABSTRACT

Objective To construct the prokaryotic expression and purification protocols for Pseudomonas aeruginosa type E flagellin (FlgE) and to study its bioactivity.Methods With analysis of Pseudomonas aeruginosa flagellin FlgE sequences,the whole-length FlgE gene was amplified from Pseudomonas aeruginosa genomic DNA by using PCR and primers with proper restriction enzyme sites.The amplified FlgE fragment and prokaryotic expression plasmid pET24a were digested with Nde Ⅰ and Hind ⅢⅣ respectively.The target fragment and vector were recovered and ligated to obtain the recombinant plasmid pET24a-FlgE.DNA sequencing of positive clone confirmed that the target gene and the junctions with vectors were all correct.The plasmid pET24a-FlgE was transformed into BL21 bacteria.The culture conditions like temperature,rotation speed,inducer concentrations,time length were optimized to achieve maximal expression of the target recombinant FlgE with 6×His tag at C terminal.FlgE-His proteins were purified using His-Trap affinity chromatography columns and identified by SDS-PAGE.The purified proteins were further subjected to endotoxin elimination with proper kits.The purified recombinant FlgE was added to cultured corneal epithelial cells for 4 h and the expression of several inflammation-related molecules was examined by using real-time quantitative PCR.Results The recombinant plasmid pET24a-FlgE was successfully constructed and high level FlgE expression was achieved in BL21 with rotation at 16℃ and 1 mmol/L isopropyl-β-D-glucopyranosyl-galactosidase induction for 20 h.Purified recombinant FlgE-His was obtained and used for primary bioactivity assay.After treatment of corneal epithelial cells with 20 μg/ml FlgE for 4 h,the expression of inflammatory cytokines IL-6,IL-8 were significantly increased.Inactivation of the FlgE with ethanol abolished its stimulatory activity.Conclusion The prokaryotic expression and purification system for recombinant Pseudomonas aeruginosa flagellin FlgE was set up,and the recombinant FlgE stimulated the expression of inflammatory factors in corneal epithelial cells.

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