Objective To construct a eukaryotic expression plasmid of pcDNA 3.1-Ag85A-CD226, and to use it as DNA vaccine then further evaluate its immunogenicity through oral administration in a mouse model.Methods The CD226-PCR2.1-ToPo plasmid was used as the template to clone CD 226 gene by PCR.The CD226 gene was then inserted into pcDNA 3.1-Ag85A plasmid to construct the recombinant plas-mid of pcDNA3.1-Ag85A-CD226.After identified by restriction enzyme analysis and sequencing , the re-combinant plasmid was transfected into HEK 293 cells by using lipofection .The expression of Ag85A-CD226 gene in HEK293 cells was detected by RT-PCR, Western blot and indirect immunofluorescence assay .The purified recombinant plasmid was used to prepare the Ag 85A-CD226 DNA vaccine by liposomal encapsula-tion.The vaccine was administered intragastrically to mice .The activities of NK cells , the cytokine levels in the supernatants of spleen cell cultures and the mRNA level of cytokines in the intestines were evaluated to analyze the immunogenicity of Ag85A-CD226 DNA vaccine.Results The Ag85A-CD226 DNA vaccine was prepared successfully .The expression of Ag85A-CD226 fusion protein was detected in HEK293 cells.The activities of NK cells from mice vaccinated with Ag 85A-CD226 DNA vaccine were higher than those from other control groups (P<0.01).The level of TNF-α, IFN-γand IL-2 in the supernatants of spleen cell cul-tures and in the intestines were significantly up-regulated in comparison with other control groups ( P <0.01).The level of IL-4 in the supernatants of spleen cell cultures was down-regulated in the experimental group (P<0.01), but the level of IL-4 in intestines showed no significant difference among the five groups (P>0.05).Conclusion The Ag85A-CD226 DNA vaccine could significantly enhance Th1 type immune responses systemically and in the intestine as in comparison with those vaccinated with single dose of Ag 85A DNA vaccine or CD226 DNA vaccine.

Trichinella spiralis,Ascaris lumbricoides,and Taenia crassiceps are all parasitic helminths that can infect humans.After invading host,the parasites can induce immune responses such as activation of immune cells and induction of cytokines in the host.The immune responses induced by parasitic helminths have similarities with anti-tumor immune responses.Recent studies have demonstrated that infection of Trichinella spiralis,Ascaris lumbricoides,or Taenia crassiceps can inhibit the development of several kinds of tumors to some extent.In this paper,the progress of anti-tumor effects induced by the above-mentioned helminths on liver cancer,lung cancer,breast cancer,malignant melanoma,myeloma,and other cancers is reviewed,which may provide a valuable reference for treatment of tumors by the immune response to helminthic infection.

Objective:To investigate the regulation and mechanism of chronic Trichinella spiralis ( Ts) infection on liver immunopathology in mice co-infected with Plasmodium berghei ANKA( PbA). Methods:According to body weight, 64 specific pathogen free female Kunming mice (6 - 8 weeks old, weighting 22 - 25 g) were divided into 4 groups by using random number table method. Control group: uninfected; Ts group: mice were mono-infected with 30 Ts larvae by oral gavage on day 0; PbA group: mice were mono-infected with 1 × 10 6PbA-infected red blood cells in 0.1 ml of phosphate buffer (PBS) administered by intraperitoneal injection on day 121; co-infected ( Ts+PbA) group: mice were infected with 30 Ts larvae by oral gavage and intraperitoneal injected with 1 × 10 6PbA-infected red blood cells in 0.1 ml PBS on day 121 after Ts infection. There were 16 mice in each group, in which 10 mice in each group were monitored for the survival rate. The peripheral red blood cell parasitemia of PbA group and Ts + PbA group were monitored every other day by light microscope examination of Giemsa-stained thin tail-blood smears from day 3 after PbA infection. Mice were sacrificed at day 135 after Ts infection and/or at day 15 after PbA infection, the mouse body weight and liver weight were measured, and the liver index were calculated. Ts-infected mice were monitored by a light microscope examination of diaphragm compression slide. Under a light microscope, the liver pathology and liver fibrosis of mice were observed and compared with hematoxylin-eosin (HE) staining and Sirius red staining, respectively. The F4/80 + Kupffer cells in liver of mice were examined by immunohistochemical staining. Results:After infection with Ts or PbA, Ts larvae cysts were observed in diaphragm tissues and PbA were observed in red blood cells under the light microscope. After PbA infection, there was no significant difference in survival rate between PbA group and Ts+ PbA group ( P > 0.05). Compared with PbA group, the peripheral red blood cell parasitemia was significantly decreased in Ts+ PbA group on days 11 and 15 after PbA infection (%: 27.104 ± 7.623 vs 45.032 ± 9.849, 60.218 ± 2.776 vs 76.778 ± 6.351, P < 0.05), and the liver index and the liver pathology score were significantly decreased in Ts+ PbA group ( P < 0.05). Sirius red staining showed that the positive area of liver fibrosis in Ts+ PbA group was significantly higher than that in PbA group ( P < 0.05). Immunohistochemical staining showed that the average optical density value of F4/80 + Kupffer cells in Ts+ PbA group was significantly higher than that in PbA group ( P < 0.01). Conclusion:Chronic Ts infection may reduce the peripheral red blood cell parasitemia, increase F4/80 + Kupffer cells expression in liver, and attenuate liver pathology in mice co-infected with PbA.

Parasitic diseases still remain the world's greatest health problems and cause huge economic burden in poor areas. The drugs currently used to treat protozoiases and helminthiases have certain defects, and it is urgent to develop more effective therapeutic drugs for these diseases. Berberine is one kind of important anti-inflammatory agents originally derived from Coptis rhizoma. The derivatives of berberine are obtained by modifying the structural site of berberine. In addition to its anti-inflammatory and anti-microbial activities, berberine and its derivatives also have significant anti-parasitic activity. In this paper, we summarized recent progress in the use of berberine and its derivatives against the infections of protozoa ( Leishmania spp ., Trypanosoma spp. , Toxoplasma gondii, Plasmodium falciparum, and Eimeria tenella) and helminths ( Schistosoma mansoni, Schistosoma. japonicum, Echinococcus granulosus, and Toxocara canis), which may providea useful reference for researchers in this field.

Objective:To investigate the therapeutic effect of artesunate on mice with acute Toxoplasma gondii ( T. gondii) infection. Methods:Based on body weight (16 - 18 g), sixty-four C57BL/6 female mice aged 6 - 8 weeks were divided into 4 groups by random number table method: uninfected control group without treatment; T. gondii-infected group (Tg group), each mouse was intraperitoneally (i.p.) infected with 100 tachyzoites of T. gondii RH strain; T. gondii-infected + artesunate treatment group (Tg + ART group), 3 hours after each mouse i.p. infected with 100 tachyzoites of T. gondii RH strain, artesunate solution was i.p. injected at a dose of 30 mg/kg, once a day for a total of 7 consecutive days; T. gondii-infected + sulfadiazine treatment group (Tg + SDZ group), 3 hours after each mouse i.p. infected with 100 tachyzoites of T. gondii RH strain, sulfadiazine solution was orally administrated at a dose of 100 mg/kg, once a day for a total of 7 consecutive days. There were 16 mice in each group, in which 10 mice were used to observe survival time and 6 mice were used to monitor body weight and collect tissue samples. Mice were weighed every day from day 1 post infection (p.i.); mice were sacrificed at day 7 p.i., the liver weights of mice were weighed and the liver indexes were calculated; liver tissues were paraffin-embedded, sectioned, and stained with hematoxylin-eosin (HE), and the pathological changes of liver tissues of mice in each group were observed under a light microscope. The expression levels of T. gondii major surface antigen 1 (SAG1) in the liver tissues of mice in each group were detected by real-time quantitative PCR for evaluating parasite load. Results:All mice in the uninfected control group were survived. The survival time was 7 - 9 days in Tg group, 8 - 11 days in Tg + ART group, and 9 - 13 days in Tg + SDZ group. Compared with Tg group, the survival times of mice in Tg + ART group and Tg + SDZ group were significantly longer ( P < 0.05). On day 7 p.i., compared with uninfected control group, Tg + ART group or Tg + SDZ group, the body weight of mice in Tg group was lower ( P < 0.05); however, there was no significant difference of body weight in Tg + ART group and Tg + SDZ group compared with uninfected control group ( P > 0.05). Compared with Tg group, Tg + ART group and Tg + SDZ group had lower liver indexes and SAG1 mRNA expression levels in the liver tissues ( P < 0.05 or < 0.001), and liver histopathological changes were milder. Compared with Tg + SDZ group, there was no significant difference in both liver index and SAG1 mRNA expression level in the liver tissue of Tg + ART group ( P > 0.05). Conclusion:Artesunate solution i.p. injection can prolong the survival time, reduce parasite load in the liver, and attenuate hepatic pathological damage, to a certain extent, of mice with acute T. gondii infection.

Objective To examine the effect of aqueous extracts of Scutellaria baicalensis Georgi and Radix paeoniae Alba on periodontitis mice and compare the results of the two herbs for the treatment of the periodontitis mice.Methods Sixty-four SPF 12-week-old male Kunming mice were selected and randomly divided into four groups:Control group (C) ;Experimental periodontitis group (P):the peridontitis models in Kunming mice were prepared by wrapping silk ligature and inoculating with putative periodontopathic bacteria; Scutellaria baicalensis Georgi treatment group(SG):periodontitis was induced by the same method described above,the mice were gavaged with Scutellaria baicalensis Georgi; Radix paeoniae Alba treatment group (RG):periodontitis was induced by the same method described above,the mice were gavaged with Radix paeoniae Alba.Four mice were sacrificed at each time point of the end of 4,6,8 and 10 weeks in each group.The histopathological changes of periodontal tissue were observed under microscope with HE staining.The level of serum IgG1 and IgG2a was measured by enzyme-linked immunosorbent assay(ELISA).Results A serious inflammatory response,alveolar progressive absorption and a large number of osteoclasts were observed in the experimental periodontitis group.However,in SG and RG,the inflammation of the periodontal tissue was decreased and tissue repair was significant.The level of serum IgG2a in SG(6 week:0.934 ± 0.006,8 week:0.743 ± 0.009,10 week:0.674 ± 0.008) and RG (6 week:1.023 ± 0.032,8 week:0.851 ± 0.032,10 week:0.790 ± 0.009) was significantly decreased after the mice were gavaged with the two herbs(P ＜ 0.01).The level of serum IgG2a in SG was significantly lower than that of RG(P ＜0.01).The level of serum IgG1 in SG(6 week:0.314 ±0.006,8 week:0.344 ± 0.004,10 week:0.367 ±0.006) and RG(6 week:0.287 ±0.005,8 week:0.303 ±0.058,10 week:0.336 ±0.006) were significantly increased(P ＜ 0.01).The level of serum IgG1 in SG was significantly higher than that of RG (P ＜ 0.0l).Conclusions Both the aqueous extracts of Scutellaria baicalensis Georgi and Radix paeoniae Alba showed therapeutic effect on periodontitis in mice.Scutellaria baicalensis Georgi was more effective than Radix paeoniae Alba.

Objective:To investigate the regulatory effect of non-specific immunity induced by Trichinella spiralis (Ts) on the immune response of small intestinal tissue of mice infected with Plasmodium berghei (Pb)ANKA. Methods:Thirty-six specific pathogen free female Kunming mice (6-8 weeks old, weighting 18-22 g) were randomly divided into 4 groups according to body weight by the random number table method, including control group, Ts-mono-infected group (Ts group), PbANKA-mono-infected group (Pb group), and Ts + Pb-co-infected group (Ts + Pb group), 9 mice in each group. The mice were fed normal food, water and normal feed. The control group was not given any experimental treatment; the Ts group was infected with 20 Ts larvae orally on the first day of the experiment; the Pb group was infected with 1 × 10 6 PbANKA erythrocytes by intraperitoneal injection on the 9th day of the experiment; the Ts + Pb group was infected with 20 Ts larvae orally on the first day of the experiment, and 1 × 10 6 PbANKA erythrocytes were given by intraperitoneal injection on the 9th day. Mice were sacrificed on 22th day after Ts infection and/or 13th day after PbANKA infection, the morphological changes of peritoneal macrophages in each group were observed by transmission electron microscope; the mRNA expression levels of M1-type macrophage markers [inducible nitric oxide synthase (iNOS) and interleukin-6 (IL-6)], M2-type macrophage markers [mannose receptor C type 2 (Mrc-2) and chitinase-like 3 (Ym1)] in the small intestinal tissue of mice in each group were detected by quantitative real-time PCR (qRT-PCR), and the ratios of mRNA expression levels of M2/M1 macrophage markers were compared. Results:Transmission electron microscope showed that the morphology and structure of peritoneal macrophages in the control were normal; more pseudopodia were observed in the peritoneal macrophages in Ts group; and more pseudopodia and engulfed Plasmodium parasites were observed in the peritoneal macrophages in Pb group and Ts + Pb group. The iNOS (1.000 ± 0.290, 1.277 ± 0.251, 3.088 ± 1.110, 2.604 ± 0.773) and IL-6 mRNA expression levels (1.000 ± 0.393, 2.180 ± 0.629, 1.650 ± 0.612, 3.242 ± 1.780) of small intestinal tissue were compared among the 4 groups, the differences were statistically significant ( F=12.420, 5.270, P < 0.05). Compared with the control group, the iNOS mRNA expression levels in the Pb and Ts + Pb groups were significantly increased ( P < 0.05); compared with the Ts group, the iNOS mRNA expression level in the Ts + Pb group was significantly increased ( P < 0.05). Compared with the control group, the IL-6 mRNA expression level in the Ts + Pb group was significantly increased ( P < 0.05). The mRNA expression levels of Mrc-2 and Ym1 of small intestinal tissue in the 4 groups were significantly different ( F=9.890, 20.500, P < 0.05). The Mrc-2 mRNA expression level in the Ts + Pb group was significantly higher than those in the control, Ts, and Pb groups ( P < 0.05). The Ym1 mRNA expression level in the Pb group was significantly higher than that in the control group ( P < 0.05); the Ym1 mRNA expression level in the Ts + Pb group was significantly higher than those in the control, Ts, and Pb groups ( P < 0.05). The Mrc-2/iNOS, Ym1/iNOS of small intestinal tissue in the 4 groups were significantly different ( F=3.642, 22.360, P < 0.05). The Mrc-2/iNOS in the Ts + Pb group was significantly higher than those in the control and Pb groups ( P < 0.05). The Ym1/iNOS in the Ts + Pb group was higher than those in the control, Ts, and Pb groups ( P < 0.05). Conclusion:The non-specific immunity induced by Ts infection is involved in the regulation of intestinal immune response of mice infected with PbANKA, which may promote M2 polarization of macrophages in the small intestinal tissue.