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As a type of highly plastic innate immune cells, macrophages may be differentiated into M1 and M2 macrophages upon different stimuli, and M2 macrophages are involved in immune regulation, tissue remodeling and regeneration, and wound healing. Previous epidemiological studies have shown a significant negative correlation between the prevalence of helminth infections and the incidence of inflammatory diseases, such as allergy and autoimmune diseases. As a common type of intestinal helminths, hookworm infection may trigger high levels of type II host immune responses, with alternative activation of macrophages, which are effective to inhibit the development and progression of inflammatory diseases. This review summarizes the advances in alternative activation of macrophages in hookworm therapy for inflammatory diseases.
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Fibroblast growth factor receptors (FGFRs) have emerged as promising targets for anticancer therapy. In this study, we synthesized and evaluated the biological activity of 66 pyrazolo[3,4-
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Soil-transmitted nematodiasis was once widely prevalent in Jiangsu Province, which seriously threatened human health and hindered socioeconomic development. The control efforts over decades resulted in a remarkable decline in the prevalence of soil-transmitted nematode human infections in Jiangsu Province, with a reduction from 59.32% in 1989 to 0.12% in 2019, and the human prevalence remains at < 0.5% since 2013. Since 1987, an integrated strategy has been adopted for the control of soil-transmitted nematodiasis in Jiangsu Province; however, the core interventions varies at different stages, which mainly include deworming, water and sanitation service improvement, health education, and monitoring and assessment. The criteria of effective soil-transmitted nematodiasis control had been achieved in all epidemic counties (districts) of Jiangsu Province by 2019. Further actions to strengthen health education and monitoring and implement precision control measures are required to consolidate the achievements of soil-transmitted nematodiasis control and eliminate the harm of soil-transmitted nematodiasis to humans. This review summarizes the epidemiology, control progress and evolution of control strategy of soil-transmitted nematodiasis in Jiangsu Province.
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Emodin is a common Chinese medicine compound with anti-inflammatory, antibacterial, anti-oxidant and lipid-lowering effects. Modern studies have found that emodin activates adenylate-activated protein kinase (AMPK) signaling molecules and regulates transcriptional factors and biological functions of relevant pathways. Nonalcoholic fatty liver disease is a chronic liver disease with a high incidence in China. With the global prevalence of obesity and metabolic syndrome, the development of nonalcoholic fatty liver disease (NAFLD) is closely related to the expression of the metabolism-related signal molecule AMPK. AMPK is a key enzyme in glycolipid metabolism that can involve different stages of NAFLD development to non-alcoholic steatohepatitis (NASH) by regulating energy metabolism in the body. In recent years, many studies have suggested that the activation of AMPK signaling molecules is related to the function realization of emodin, and lipid synthesis, fatty acid oxidation, insulin sensitivity and mitochondrial function-related transcription factors affected by AMPK downstream signaling molecules and other biological effects can be interacted with each other. The detailed mechanism of action associated with AMPK activation provides new thought about the treatment of NAFLD by emodin. This paper mainly summarizes the research progress of emodin by participating in the various stages of NAFLD by AMPK-related signaling pathways through literature retrieval and comprehensive analysis. It lays a foundation for further research on the therapeutic effect and mechanism of emodin on NAFLD.
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Objective To investigate the polarization of human acute monocytic leukemia THP-1 cells-derived macrophages induced by Nippostrongylus brasiliensis proteins in vitro, so as to provide insights into the elucidation of the mechanisms underlying host immune responses to hookworm infections. Methods The in-vitro culture of N. brasiliensis was established and maintained in the laboratory, and the third- (L3) and fifth-stage larvae (L5) were collected under a sterile condition for preparation of L3 and L5 proteins. The in-vitro culture of THP-1 cells was established, stimulated with 500 ng/mL PMA to yield M0 macrophages that were adherent to the plate wall. The LPS + IFN-γ group, IL-4 + IL-13 group, L3 protein group and L5 protein group were given stimulation with 500 ng/mL LPS plus 100 ng/mL IFN-γ, IL-4 and IL-13 (both 100 ng/mL), L3 protein (5 mg/mL) and L5 protein (5 mg/mL), respectively, while the negative control group was given no stimulation. The cell morphology was observed using microscopy, the mRNA expression of M1/M2 macrophages-specific genes was quantified using a quantitative real-time PCR (qPCR) assay, and the surface markers of M1/M2 macrophages were detected using flow cytometry, while the levels of cytokines secreted by M1/M2 macrophages were measured using enzyme-linked immunosorbent assay (ELISA) following stimulations, so as to examine the polarization of THP-1-derived macrophages induced by N. brasiliensis proteins in vitro. Results Following stimulation with PMA, THP-1 cells appeared wall-adherent M0 macrophages, and polarized to typical M1 macrophages following stimulation with LPS + IFN-γ, and typical M2 macrophages following stimulation with IL-4 + IL-13, IL-3 protein or L5 protein. There was a significant difference in the proportion of M1 macrophages among the negative control group, the LPS + IFN-γ group, the IL-4 + IL-13 group, the L3 protein group and the L5 protein group (χ2 = 3 721.00, P < 0.001), with the highest proportion detected in the LPS + IFN-γ group, and there was also a significant difference in the proportion of M2 macrophages among groups (χ2 = 105.43, P < 0.001). There were significant differences among groups in terms of the mRNA expression of CCL2 (F = 191.95, P < 0.001), TNF-α (F = 129.95, P < 0.001), IL-12b (F = 82.89, P < 0.001), PPARγ (F = 11.30, P < 0.001), IL-10 (F = 9.51, P < 0.001) and Mrc1 genes (F = 12.35, P < 0.001). In addition, there were significant differences in the proportion of positive CD86 and CD206 expression among groups (χ2 = 24 004.33 and 832.50, P < 0.001). Higher IL-1β and TNF-α levels were measured in the LPS + IFN-γ group than in the IL-4 + IL-13 group, the L3 protein group and the L5 protein group (P < 0.001), and greater TGF-β1 and IL-10 levels were seen in the IL-4 + IL-13 group, the L3 protein group and the L5 protein group than in the negative control group and the LPS + IFN-γ group (P < 0.05). Conclusions Both L3 and L5 proteins of N. brasiliensis may induce the polarization of THP-1-derived macrophages to M2 type in vitro.
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Objective To establish a recombinase-aided isothermal amplification (RAA) assay for the nucleic acid detection of Angiostrongylus cantonensis. Methods The internal transcribed spacer-1 (ITS1) gene sequence of A. cantonensis was used as the detection target sequence, and the specific primers and probes were designed and synthesized, followed by screening of the primers and probes with the highest specificity, to establish the basic and fluorescent RAA assay for nucleic acid detection of A. cantonensis. The sensitivity of the fluorescent RAA assay was evaluated by using the target gene fragment sequence-contained recombinant plasmids at various copy numbers and the genomic DNA from A. cantonensis as the template DNA samples, and the specificity of the fluorescent RAA assay was evaluated by using the genomic DNA from A. cantonensis, Schistosoma mansoni, Ascaris lumbricoides, Clonorchis sinensis, Echinococcus granulosus and Ancylostoma duodenale, as well as Pomacea canaliculata and Biomphalaria straminea snail tissues as the template DNA samples. Results A fluorescent RAA assay was successfully established for nucleic acid detection of A. cantonensis, which achieved real-time amplification of the specific DNA fragment of A. cantonensis within 20 min at 37 ℃. By using the target gene fragment sequence-contained recombinant plasmids at various copy numbers and the genomic DNA from A. cantonensis as the DNA templates, the lowest detection limits of the fluorescent RAA assay were 10 copies/μL of recombinant plasmids and 100 pg/μL of genomic DNA, respectively. The fluorescent RAA assay was negative for detection of the genomic DNA from A. cantonensis, S. mansoni, A. lumbricoides, C. sinensis, E. granulosus, A. duodenale, and P. canaliculata and B. straminea snail tissues. Conclusions A simple, rapid fluorescent RAA assay has been successfully established, which has a high sensitivity and specificity for the nucleic acid detection of A. cantonensis.
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Objective To establish a novel nucleic acid assay for detection of Giardia lamblia based on the recombinase-aided isothermal amplification (RAA) assay, and evaluate its sensitivity and specificity for detection of G. lamblia. Methods The specific primer sequences and florescent probes were designed and synthesized based on the G. lamblia β-giardin gene as the target gene, and a fluorescent RAA assay was established. The recombinant plasmids at various copies (containing the β-giardin gene target sequence) and the genomic DNA of G. lamblia at various concentrations were used as templates for the fluorescent RAA assay to assess the sensitivity, and the genomic DNA from G. lamblia, Schistosoma japonicum, Clonorchis sinensis, Cryptosporidium parvum, Ascaris lumbricoides, Salmonella and Shigella was used as templates to assess the specificity of the fluorescent RAA assay. Results A novel fluorescent RAA assay was successfully established for detection of G. lamblia, which allowed the rapid and specific amplification of the target gene fragments at 39 ℃ within 20 min. The sensitivities of the fluorescent RAA assay were 102 copies/μL and 1 pg/μL for detection of the recombinant plasmid and G. lamblia genomic DNA, respectively, and the fluorescent RAA assay was negative for detection of the genomic DNA from S. japonicum, C. sinensis, C. parvum, A. lumbricoides, Salmonella and Shigella, which showed a high specificity. Conclusions A fluorescent RAA assay, which is simple, sensitive and specific, is successfully established for nucleic acid detection of G. lamblia.
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Objective To establish a nucleic acid assay for detection of Echinococcus granulosus based on recombinase-aided isothermal amplification (RAA) assay. Methods The 12S rRNA gene of E. granulosus was selected as the target gene, and the specific primers and fluorescent probes for RAA assay were designed, screened and synthesized to establish a fluorescent RAA assay for detection of E. granulosus. The sensitivity of the fluorescent RAA assay was evaluated using different copy numbers of target gene sequence-contained recombinant plasmids and various concentrations of E. granulosus genomic DNA as templates, and the specificity of the fluorescent RAA assay was evaluated using the genomic DNA from E. granulosus, E. multilocularis, Schistosoma japonicum, S. mansoni, Ancylostoma duodenale, Clonorchis sinensis, Taenia saginata, Spirometra mansoni and Taenia solium as templates. Results A fluorescent RAA assay was successfully established for detection of E. granulosus, which achieved specific amplification of E. granulosus genomic DNA within 20 min at 39 ℃. The lowest detection limit of the fluorescent RAA assay was 10 copies/μL of recombinant plasmids and 0.1 ng/μL E. granulosus genomic DNA, which exhibited a high sensitivity, and the fluorescent RAA assay was all negative for the genomic DNA from E. multilocularis, S. japonicum, S. mansoni, A. duodenale, C. sinensis, T. saginata, Spirometra mansoni and T. solium, which exhibited a high specificity. In addition, this fluorescent RAA assay successfully detected genomic DNA from E. granulosus cysts. Conclusions A rapid, sensitive and specific fluorescent RAA assay is successfully established for nucleic acid detection of E. granulosus.
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Objective To investigate the risk of Anisakis infections among high-risk populations along the coastal areas of Jiangsu Province, so as to develop the strategy for the prevention and control of anisakiasis in the province. Methods Three counties along the coastal areas of Jiangsu Province were selected as the study sites in 2018, including Rudong County in Nantong City, Haizhou District in Lianyungang City and Dongtai City in Yancheng City. The knowledge, attitude and practice (KAP) of anisakiasis prevention and control, and the prevalence of serum specific IgG antibody against Anisakis were investigated among high-risk populations among these three study sites, including fishermen, fish seller and people who liked eating fresh and live marine fish. Factors affecting the prevalence of the specific IgG antibody against Anisakis were identified using a multiple logistic regression model. In addition, Anisakis larvae infections were detected in fresh and live marine fish samples collected from local markets, and the prevalence and intensity of Anisakis infections were estimated. Results A total of 625 high-risk populations were investigated, including 349 men (55.8%). Only 13.0% of the subjects heard about anisakiasis, and a low awareness rate of anisakiasis prevention and control knowledge was seen among these three types of high-risk populations. There were 21.6% of the subjects eating raw or half-cooked marine fish, 5.8% eating undercooked marine fish, 3.2% presenting vomiting, nausea and diarrhea after eating marine fish, 5.1% developing systemic allergic symptoms, and 65.6% using the same chopping board for raw and cooked food. The sero-prevalence of the anti-Anisakis IgG antibody was 7.0% among the study subjects. Multiple logistic regression analysis identified education level [OR = 0.687, 95% CI (0.478, 0.987)] and development of systemic allergic symptoms [OR = 4.641, 95% CI(1.411, 15.268)]as factors affecting the positive anti-Anisakis IgG antibody among the study subjects. Among 494 fresh and live marine fish detected, the prevalence and intensity of Anisakis larvae infection was 64.0% and 8.1 larvae per fish, with high prevalence seen in Trichiurus haumela and Pneumatophorus japonicas. Conclusions The awareness of anisakiasis prevention and control knowledge is low among the high-risk populations living along the coastal areas of Jiangsu Province, and there are high-risk behaviors, such as eating raw or half-cooked food, using the same chopping board for raw and cooked food. In addition, the prevalence of Anisakis infections is high in the marine fish in these areas. Therefore, the health education and health promotion for anisakiasis prevention and control should be intensified.
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Objective: To observe the effect of puerarin on phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and glycogen synthase kinase-3β (GSK-3β) in insulin resistant HepG2 cells. Method: HepG2 cells were treated with palmitic acid 0.5 mmol·L-1 and insulin 9×10-4 U·L-1 to induce insulin resistant condition for 24 h. Cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay to determine the concentration of puerarin. This experiment included normal control group, model control group and puerarin groups of different doses (40, 80, 160,320 μg·L-1). Glucose detection kit was used to detect the content of glucose in cell culture supernatant. Tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) levels in supernatant of cell culture medium were detected by enzyme-linked immunosorbent assay (ELISA). Hepatic glycogen assay kit was used for detecting the hepatic glycogen content in HepG2 cells. Western blot was applied to detect protein expression levels of PI3K, Akt, p-Akt, GSK-3β and p-GSK-3β. Result: Compared with those in the normal control group, the glucose consumption rate was significantly down-regulated in HepG2 cells in the model control group (PPα and IL-6 were increased in supernatant of cell culture medium (PPβ protein expression was up-regulated (PPα and IL-6 were reduced in supernatant of cell culture medium (PPβ protein expression was down-regulated, but its phosphorylation inactivation was increased (PConclusion: Puerarin alleviates the insulin resistance of HepG2 cells by strengthening the PI3K/Akt/GSK-3β signal transduction process and increasing the glycogen content in hepatocytes.
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Hookworm infection remains a global health concern, which threatens human health. Hookworm infection is widely prevalent across the world, notably in tropical and subtropical areas. Recently, with the in-depth study of the immunity of parasitic infections, the“bidirectional effect”of host immune responses induced by helminth infections (including hookworm infections) has become increasingly prominent. On one hand, an immune response is induced in the host to kill the infected worms; on the other hand, the host produces a series of immunological changes that are conducive to the maintenance of parasite survival. The immune state of the host is regulated by various complicated mechanisms, and this may lead to the reduction in the incidence of allergic and autoimmune diseases or alleviation of the disease symptoms, which provide new insights into the management of these allergic and autoimmune diseases. The present article reviewed the advances of host immune responses induced by hook-worm infection and its potential values in the treatment of allergic asthma, inflammatory bowel disease and rheumatoid arthritis.
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Objective To establish a recombinase aided isothermal amplification (RAA) assay for detection of Clonorchis sinensis. Methods The 18S ribosomal RNA (18S rRNA) sequence of C. sinensis was used as the target sequence, and specific primers and probes were designed, synthesized and screened to establish a rapid fluorescent RAA assay for the detection of C. sinensis. Then, the sensitivity of the fluorescent RAA assay was evaluated using the recombinant plasmids containing various copy numbers of DNA fragments and C. sinensis genomic DNA at various concentrations, and the specificity of the fluorescent RAA as say was evaluated using the genomic DNA of Ascaris lumbricoides, Echinococcus granulosus, Schistosoma japonicum, Ancylostoma duodenale and S. mansoni as templates. DNA samples were extracted from the feces containing C. sinensis eggs and freshwater fish containing metacercaria for the fluorescent RAA assay, and the performance for detection of C. sinensis-infected samples was preliminarily assessed in the field. Results A fluorescent RAA assay for detection of C. sinensis was successfully established, which was feasible for specific amplification of C. sinensis genomic DNA at 39 °C within 20 min. The lowest detection limit was 10 copies/μL if the recombinant plasmid containing various copy numbers of DNA fragments was used as a template, and the lowest detection limit was 3 pg/μL if the C. sinensis genomic DNA at various concentrations served as a template. All detections were negative if the genomic DNA of A. lumbricoides, E. granulosus, S. japonicum, A. duodenale, and S. mansoni was used as templates. In addition, the fluorescent RAA assay showed a high performance for the detection of C. sinensis-infected samples in the field, which successfully detected C. sinensis-infected human and rat fecal samples and Pseudorasbora parva samples. Conclusion A fluorescent RAA assay is successfully established, which is simple, rapid, sensitivity and specific for detection of C. sinensis.
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Objective To establish a recombinase-aided isothermal amplification (RAA) assay for detection of Cryptosporidium. Methods Based on Cryptosporidium-specific 18S rRNA selected as the target gene to be detected, and the primer sequences and fluorescent probes designed using the software Amplfix, and a fluorescent RAA assay was established and optimized. The fluorescent RAA assay was performed to detect 18S RNA target sequence-contained recombinant plasmids at various copies, genomic DNA of Cryptosporidium oocysts at various concentrations, and genomic DNA extracted from various numbers of Cryptosporidium oocysts to assess the sensitivity of the assay, and to detect genomic DNA extracted from Cryptosporidium oocysts, Giardia lamblia cysts, Schistosoma japonicum eggs, Ascaris lumbricoides eggs, Clonorchis sinensis eggs, Salmonella and Shigella to determine the specificity of the assay. Results A fluorescent RAA assay was successfully established, which was effective to amplify the specific 18S RNA gene fragments of Cryptosporidium within 20 min at 39 ℃. The lowest limits of the fluorescent RAA assay were 102 copies/μL for detection of 18S RNA target sequence-contained recombinant plasmids at various copies, 1 pg/μL for detection of genomic DNA of Cryptosporidium oocysts at various concentrations, and one Cryptosporidium oocyst/μL for detection of genomic DNA extracted from various numbers of Cryptosporidium oocysts, and the fluorescent RAA assay was all negative for detection of genomic DNA from G. lamblia cysts, S. japonicum eggs, A. lumbricoides eggs, C. sinensis eggs, Salmonella and Shigella. Conclusion A novel fluorescent RAA assay is successfully established, which is simple, rapid, sensitive and specific to detect genomic DNA of Cryptosporidium oocysts.
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Hookworm infection remains a global health concern, which threatens human health. Hookworm infection is widely prevalent across the world, notably in tropical and subtropical areas. Recently, with the in-depth study of the immunity of parasitic infections, the“bidirectional effect”of host immune responses induced by helminth infections (including hookworm infections) has become increasingly prominent. On one hand, an immune response is induced in the host to kill the infected worms; on the other hand, the host produces a series of immunological changes that are conducive to the maintenance of parasite survival. The immune state of the host is regulated by various complicated mechanisms, and this may lead to the reduction in the incidence of allergic and autoimmune diseases or alleviation of the disease symptoms, which provide new insights into the management of these allergic and autoimmune diseases. The present article reviewed the advances of host immune responses induced by hook-worm infection and its potential values in the treatment of allergic asthma, inflammatory bowel disease and rheumatoid arthritis.
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Objective To establish a recombinase aided isothermal amplification (RAA) assay for detection of Clonorchis sinensis. Methods The 18S ribosomal RNA (18S rRNA) sequence of C. sinensis was used as the target sequence, and specific primers and probes were designed, synthesized and screened to establish a rapid fluorescent RAA assay for the detection of C. sinensis. Then, the sensitivity of the fluorescent RAA assay was evaluated using the recombinant plasmids containing various copy numbers of DNA fragments and C. sinensis genomic DNA at various concentrations, and the specificity of the fluorescent RAA as say was evaluated using the genomic DNA of Ascaris lumbricoides, Echinococcus granulosus, Schistosoma japonicum, Ancylostoma duodenale and S. mansoni as templates. DNA samples were extracted from the feces containing C. sinensis eggs and freshwater fish containing metacercaria for the fluorescent RAA assay, and the performance for detection of C. sinensis-infected samples was preliminarily assessed in the field. Results A fluorescent RAA assay for detection of C. sinensis was successfully established, which was feasible for specific amplification of C. sinensis genomic DNA at 39 °C within 20 min. The lowest detection limit was 10 copies/μL if the recombinant plasmid containing various copy numbers of DNA fragments was used as a template, and the lowest detection limit was 3 pg/μL if the C. sinensis genomic DNA at various concentrations served as a template. All detections were negative if the genomic DNA of A. lumbricoides, E. granulosus, S. japonicum, A. duodenale, and S. mansoni was used as templates. In addition, the fluorescent RAA assay showed a high performance for the detection of C. sinensis-infected samples in the field, which successfully detected C. sinensis-infected human and rat fecal samples and Pseudorasbora parva samples. Conclusion A fluorescent RAA assay is successfully established, which is simple, rapid, sensitivity and specific for detection of C. sinensis.
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Objective To establish a recombinase-aided isothermal amplification (RAA) assay for detection of Cryptosporidium. Methods Based on Cryptosporidium-specific 18S rRNA selected as the target gene to be detected, and the primer sequences and fluorescent probes designed using the software Amplfix, and a fluorescent RAA assay was established and optimized. The fluorescent RAA assay was performed to detect 18S RNA target sequence-contained recombinant plasmids at various copies, genomic DNA of Cryptosporidium oocysts at various concentrations, and genomic DNA extracted from various numbers of Cryptosporidium oocysts to assess the sensitivity of the assay, and to detect genomic DNA extracted from Cryptosporidium oocysts, Giardia lamblia cysts, Schistosoma japonicum eggs, Ascaris lumbricoides eggs, Clonorchis sinensis eggs, Salmonella and Shigella to determine the specificity of the assay. Results A fluorescent RAA assay was successfully established, which was effective to amplify the specific 18S RNA gene fragments of Cryptosporidium within 20 min at 39 ℃. The lowest limits of the fluorescent RAA assay were 102 copies/μL for detection of 18S RNA target sequence-contained recombinant plasmids at various copies, 1 pg/μL for detection of genomic DNA of Cryptosporidium oocysts at various concentrations, and one Cryptosporidium oocyst/μL for detection of genomic DNA extracted from various numbers of Cryptosporidium oocysts, and the fluorescent RAA assay was all negative for detection of genomic DNA from G. lamblia cysts, S. japonicum eggs, A. lumbricoides eggs, C. sinensis eggs, Salmonella and Shigella. Conclusion A novel fluorescent RAA assay is successfully established, which is simple, rapid, sensitive and specific to detect genomic DNA of Cryptosporidium oocysts.
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Objective To optimize the process conditions of the oil extraction ofseabuckthorn oil and to evaluate its antioxidant activity by anti-free radical action.Methods The extraction time and particle size of sea-buckseed oil were optimized by using the response surface software design-expert.Its antio xidant activity was studied through its anti-dpph free radical reaction.Results The best process of seabuckthorn seed oil was extracting time 3 h,material liquid than 1:8,extraction temperature 80 C,about 30 mesh size,the yield is highest at 11.13％.The optimum reaction time was 8 min in control,and with the increase of concentration,seabuckthorn oil antioxidant activity increased,when the addition amount of 4.00 ml sample,clearance rate as high as 77.62％.Conclusions This method is simple and reliable,the extraction rate is high,and the test results show that the oil has obvious anti-oxidative effect,which can be used as whitening and wrinkling products to delay the aging of human body.
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Objective To report the surgical nursing strategy of a patient undergoing resection of a rare giant neurofibroma.Methods Through the preoperative comprehensive assessment of patients,multidisciplinary joint consultation and proposed possible surgical risks and countermeasures,specialist to formulate surgical plans,surgical care and nursing key points,difficulties and countermeasures.Results The psychological nursing,detailed and perfect the preoperative preparation work,the personnel allocation was sufficient,the division of labor was clear,the disease changes were actively observed in the operation,the venous pathway was strictly managed,the accurate record,the risk that might occur in the active prevention operation and the treatment plan,the postoperative closely monitors the condition.Good observation and nursing in the operation area to prevent infection.Conclusions In view of difficult and complicated operation,through the comprehensive formulation of surgical nursing measures and strict execution in surgery,it is the guarantee for the smooth completion of the operation,the operation process is smooth,the successful implementation of tumor resection,the effect is satisfactory,the patient is returned to the ward,and there is no special discomfort.
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Objective To establish a novel method for the detection of Schistosoma japonicum specific gene fragments by re-combinase aided isothermal amplification(RAA).Methods The gene fragment SjG28 of S.japonicum was selected as the tar-get gene fragment to be detected,and the primers were designed according to the mechanism of RAA reaction.The reaction of isothermal amplification of S.japonicum was established and optimized.Then this method was applied to amplify and detect the specific gene fragment in the gradient diluent SjG28-recombiant plasmids and different concentrations of S.japonicum genomic DNA to estimate the sensitivity of this method.The samples were also detected by polymerase chain reaction(PCR)in parallel as control.This method was applied to detect the genomic DNA of S.mansoni,Ascaris lumbricoides,and Ancylostoma duodenale to evaluate the specificity.Results The specific gene fragment was amplified from genomic DNA of adult worms and eggs of S.japonicum by recombinase aided isothermal amplification reaction established in this study.The reaction can be completed with-in 30 minutes and the minimum detectable template was 20 copies of plasmids or 0.5 ng of genomic DNA per microliter.Other parasites'genomic DNAs,such as S.mansoni,A.lumbricoides,An.duodenale and healthy human blood genomic DNA were not able to be detected by this method.Conclusion A novel method for the detection of S.japonicum specific gene fragments by re-combinase aided isothermal amplification is established in this study,which can be carried out conveniently and rapidly with a considerable sensitivity and specificity,showing the prospect for application in the diagnosis of schistosomiasis japonica.
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Objective To understand the current status of chronic filariasis patients in Jiangsu Province so as to provide basic data for following-up care for them. Methods The patients were followed up one by one according to history archives between June and July, 2018, and the clue investigation was also conducted. The base data of the patients was collected through a face-to-face questionnaire survey and analyzed. Results There were still 3 160 chronic filariasis patients in Jiangsu Province. Among them, the male accounted for 40.0%, and 91.8% of the patients were older adults aged 60 years or above. From the aspect of regional distribution, Suqian (24.2%), Huai’an (19.5%), Suzhou (17.3%), Xuzhou (11.2%), and Yancheng (9.8%) were the five top high prefectures. The patients with simple lymphatic inflammation or lymphadenitis, simple lymphedema or elephantiasis, simple chyluria, simple hydrocele of tunica vaginalis, and two symptoms or more accounted for 2.7%, 37.1%, 11.2%, 0.9%, and 48.1%, respectively. For the patients with lymphedema or elephantiasis, 97.8% of edema was seen in the lower limbs, and more than 90% of the edema stages were I-III. The number of current caring sites was 220, covering 2 091 patients. The average number of times of caring activities in this year was 3.2. The average cumulative time of caring activities among all the sites was 11.3 years. Conclusions The number of chronic filariasis patients has been dramatically decreased, most of the patients are old and have long disease durations. The caring sites have not covered all the patients. In order to release the symptoms and improve the life quality of the patients, all the patients should be taken care of in Jiangsu Province.