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Tao Z.-Y.,Bengbu Medical College | Tao Z.-Y.,Anhui Key Laboratory of Infection and Immunity | Xu S.,China Institute of Technology | Wang Y.-Y.,Bengbu Medical College | And 6 more authors.
Chinese Journal of Schistosomiasis Control | Year: 2014

Objective: To establish a method based on repetitive protein sequences and linear B cell epitope to predict and screen specific peptides of Plasmodium vivax. Methods: A P. vivax protein sequence database was reconstructed based on PlasmoDB data, and a customized software for searching of repetitive sequences was used to count the repetition times of each 16 aa peptide in the whole database, and the highly repetitive peptides were chosen to predict the potential linear B cell epitopes. The repetitive peptides with P. vivax specificity were selected as candidate antigen peptides to synthesize and to couple with KLH carrier protein for immunizing BALB/c mice. After the immunization, the antibody titers of the immunized mice were detected. Results: The repetitive information of 16 aa peptides was analyzed by screening of the total 5 432 peptide sequences in the P. vivax database. A total of 22 peptides were identified as candidate peptides from the top 1 000 repetitive peptides by linear B cell epitope prediction on the BcePred website. Through clustering analysis and similarity comparison, five potential P. vivax specific peptides were selected, synthesized and then coupled with KLH to immunize the mice. The antibody titers of the immunized mice induced by the 5 peptides were all above 1 : 9 000. Conclusion: The method for predicting and screening of specific peptides of P. vivax based on repetitive protein sequences and linear B cell epitope has been established successfully, and all the 5 peptides obtained by the method can induce the high titer antibody in mice.


Wang X.-M.,Bengbu Medical College | Wang X.-M.,Anhui Key Laboratory of Infection and Immunity | Xia L.-H.,Centers for Disease Control and Prevention | Fang Q.,Bengbu Medical College | And 9 more authors.
Chinese Journal of Schistosomiasis Control | Year: 2013

Objective: To understand the malaria epidemiologic characteristics in Wuhe County, Anhui Province from 2009 to 2011, so as to provide the evidence for formulating effective malaria control and prevention interventions. Methods: The data of malaria cases from the reporting system and malaria epidemiological investigations were collected and analyzed statistically in Wuhe County from 2009 to 2011. Results: The total number of malaria cases was 349 in Wuhe County from 2009 to 2011, and the incidence showed a downtrend. The sex ratio of patients was 1.42:1 (205 males vs. 144 females). The ranks of patient age groups were 11-20 years old at the first and 31-40 at the second, and the youngest was 1 year old, and the oldest was 100 years old. The seasonal factor was observed clearly, malaria cases were rare during the period of January to March, the case numbers increased in April and reached the peak during the period of June to September, and the cases from November to December accounted for only 2.01% of the total cases. Among the 349 patients, there were only 2 patients (0.57%) living in urban areas and the rest 347 patients (99.43%) all living in rural areas. The incidence of the urban population was 0.08/10000, and it was significantly lower than that of the rural population (1.78/10000, P < 0.05). The highest incidences occurred in hill townships, Zhuding Town and Xiaoxi Town, the annual incidences were 5.53/10000 and 4.78/10 000, respectively. The cases in hill townships accounted for 36.10% of the total cases. The most patients in these areas lived in brick or cement buildings without mosquito-proof doors or windows. They preferred sleeping outside during summer, and were generally lack of the malaria prevention knowledge. Conclusion: In Wuhe County, the malaria incidence is decreasing year by year, and the epidemiologic factors are related to the living conditions, mosquito-proof facility using, sleeping habit, and malaria awareness of the residents.


Wang X.-M.,Bengbu Medical College | Wang X.-M.,Anhui Key Laboratory of Infection and Immunity | Luo J.-K.,Anhui Key Laboratory of Infection and Immunity | Luo J.-K.,Bengbu Medical College | And 11 more authors.
Chinese Journal of Schistosomiasis Control | Year: 2014

Objective: To construct recombinant Mycobacterium smegmatis vaccine expressing Cysticercus cellulosae cC1 antigen. Methods: The recombinant pET28a-cC1 plasmid was extracted and double digested by Xho I and BamH I restriction enzymes, and shuttle plasmid pMV261 was extracted and double digested by Hind III and BamH I restriction enzymes. Both fragments were modified by Klenow fragment to form blunt end, then the large fragments of cC1 and pMV261 plasmid were purified and ligated by T4 ligase enzyme. The recombinant pMV261-cC1 plasmid was constructed and sequenced. Then the pMV261-cC1 plasmid was transformed into Mycobacterium smegmatis by the electrotransformation method. The recombinant cC1? Mycobacterium smegmatis was induced by heat and identified by the Western blotting method with the sera of cysticercosis patients. In addition, the growth states of the Mycobacterium smegmatis and the recombinant cC1-Mycobacterium smegmatis were compared and the growth curves were drawn. Results: The restriction enzyme and sequencing results showed that the recombinant pMV261-cC1 plasmid was successfully constructed. After heat induction, a 40 kD band was showed by PAGE analysis of cC1-Mycobacterium smegmatis. The Western blotting results showed that the sera of cysticercosis patients could recognize the 40 kDa band, which suggested that cC1 protein was expressed in Mycobacterium smegmatis. Compared with the Mycobacterium smegmatis, the recombinant cC1-Mycobacterium smegmatis showed no significant difference in proliferation characteristics. Conclusion The recombinant cC1-Mycobacterium smegmatis vaccine has been successfully constructed.


Liu D.,Bengbu Medical College | Liu D.,Anhui Key Laboratory of Infection and Immunity | Xia H.,Bengbu Medical College | Xia H.,Anhui Key Laboratory of Infection and Immunity | And 12 more authors.
Chinese Journal of Schistosomiasis Control | Year: 2012

Objective: To clone a Plasmodium vivax Duffy binding protein critical functional region II (PvDBPII) gene of central China isolate, and to express and identify the recombinant PvDBPII protein in vitro. Methods: PCR was performed to amplify PvDBPII from P. vivax DNA of a central China isolate and the PCR product was inserted into pET28a(+) vector. pET28a-PvDBPII recombinant plasmid was constructed and transformed into E. coli host BL21 (DE3+). IPTG was used to induce the recombinant PvDBPII protein fused with His tag, and the protein was purified by His-NTA affinity chromatography. The recombinant protein was identified by SDS-PAGE and Western blot. Results: The PCR product of PvDBPII gene was about 1.1 kb, meeting the expectation of predicted fragment size. The recombinant pET28a-PvDBPII plasmid was verified by sequencing that the insertion was correct both in direction and in frame, but with 4 non-synonymous mutations compared to reference P. vivax strain Sal-I. SDS-PAGE, and Western blot analysis showed that the recombinant PvDBPII protein was about 44 kDa, and could be recognized by pooled sera from vivax malaria patients. Conclusion: The PvDBPII gene of central China isolate is successfully cloned, and recombinant PvDBPII is expressed, thereby providing opportunity for further study on PvDBPII.


Jiang L.,Anhui Key Laboratory of Infection and Immunity | Yao C.,Bengbu Medical College | Jin Q.,Bengbu Medical College | Li B.,Bengbu Medical College
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology | Year: 2014

To investigate the effect of interleukin (IL)-17 on the apoptosis of neutrophils (PMNs) from peripheral blood of patients with pulmonary tuberculosis (TB) and explore the possible involved signaling pathway. The fresh isolated PMNs from peripheral blood of TB patients and healthy adults were cultured for 0-24 hours, and then stained with annexin V. The flow cytometry was used to measure cell apoptosis of PMNs. The fresh isolated PMNs were cultured with different concentrations of IL-17 for 24 hours, with or without pretreatment of MAPK inhibitor U0126 for 30 minutes, the apoptosis of PMNs was detected by flow cytometry. The apoptosis of PMNs in both TB patients and healthy adults increased with the culture time going on (P<0.05). The spontaneous apoptotic (annexin V+) rates of PMNs at 0, 6, 12 and 24 hours of culture time in TB patients were (4.49 ± 1.39)%, (21.89 ± 2.90)%, (39.96 ± 4.15)%, and (68.35 ± 7 .01)%, respectively, which were significantly higher than the corresponding ones in normal healthy groups, (2.65 ± 0.75)%, (11.00 ± 1.72)%, (25.84 ± 3.90)%, and (45.59 ± 4.10)%, respectively (P<0.05). After cultured with IL-17 for 24 hours, the apoptotic rates of PMNs in TB patients and healthy adults were respectively (59.81 ± 7.19)% and (34.65 ± 4.79)% in the group of IL-17 at 0.5 μg/L, and (51.62 ± 6.91)% and (29.04 ± 3.62)% in the group of IL-17 at 5 μg/L; they were significantly lower than those in control groups without IL-17, (68.35 ± 7.01)% and (45.59 ± 4.10)%, respectively, (P<0.05). However, in the group of IL-17 at 50 μg/L, the apoptosis rates of PMNs in TB patients and healthy adults were (76.04 ± 5.59)% and (53.24 ± 4.62)%, respectively, which were obviously higher than those of the control groups without IL-17 (P<0.05). Pretreatment of PMNs with U0126 mostly inhibited the anti-apoptosis effect of IL-17 at 5 μg/L on PMNs of TB patients and normal subjects. The apoptosis of PMNs in both TB patients and healthy adults increases along with the culture time, and the apoptosis rate of PMNs in TB patients is higher than that of healthy adults. The lower concentrations of IL-17 can delay, but the higher concentration of Il-17 can accelerate the apoptosis of PMNs in TB patients. The inhibitory role of IL-17 in apoptosis of PMNs may be involved in the ERK pathway.


PubMed | Bengbu Medical College and Anhui Key Laboratory of Infection and Immunity
Type: Journal Article | Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology | Year: 2014

To investigate the effect of interleukin (IL)-17 on the apoptosis of neutrophils (PMNs) from peripheral blood of patients with pulmonary tuberculosis (TB) and explore the possible involved signaling pathway.The fresh isolated PMNs from peripheral blood of TB patients and healthy adults were cultured for 0-24 hours, and then stained with annexin V. The flow cytometry was used to measure cell apoptosis of PMNs. The fresh isolated PMNs were cultured with different concentrations of IL-17 for 24 hours, with or without pretreatment of MAPK inhibitor U0126 for 30 minutes, the apoptosis of PMNs was detected by flow cytometry.The apoptosis of PMNs in both TB patients and healthy adults increased with the culture time going on (P<0.05). The spontaneous apoptotic (annexin V) rates of PMNs at 0, 6, 12 and 24 hours of culture time in TB patients were (4.49 1.39)%, (21.89 2.90)%, (39.96 4.15)%, and (68.35 7 .01)%, respectively, which were significantly higher than the corresponding ones in normal healthy groups, (2.65 0.75)%, (11.00 1.72)%, (25.84 3.90)%, and (45.59 4.10)%, respectively (P<0.05). After cultured with IL-17 for 24 hours, the apoptotic rates of PMNs in TB patients and healthy adults were respectively (59.81 7.19)% and (34.65 4.79)% in the group of IL-17 at 0.5 g/L, and (51.62 6.91)% and (29.04 3.62)% in the group of IL-17 at 5 g/L; they were significantly lower than those in control groups without IL-17, (68.35 7.01)% and (45.59 4.10)%, respectively, (P<0.05). However, in the group of IL-17 at 50 g/L, the apoptosis rates of PMNs in TB patients and healthy adults were (76.04 5.59)% and (53.24 4.62)%, respectively, which were obviously higher than those of the control groups without IL-17 (P<0.05). Pretreatment of PMNs with U0126 mostly inhibited the anti-apoptosis effect of IL-17 at 5 g/L on PMNs of TB patients and normal subjects.The apoptosis of PMNs in both TB patients and healthy adults increases along with the culture time, and the apoptosis rate of PMNs in TB patients is higher than that of healthy adults. The lower concentrations of IL-17 can delay, but the higher concentration of Il-17 can accelerate the apoptosis of PMNs in TB patients. The inhibitory role of IL-17 in apoptosis of PMNs may be involved in the ERK pathway.

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