Trichomonas vaginalis triggers the release of THP-1 extracellular traps.

Trichomonas vaginalis is responsible for the prevalence of trichomoniasis, which may be one of the most epidemic nonviral sexually transmitted pathogens. Extracellular traps (ET) are a unique form of innate immunity against infection; they bind to and kill microorganisms. However, the effect of T. vaginalis on ET release in the human monocytic cell line THP-1 remains unclear. In the present study, the morphology of ET derived from THP-1 in response to T. vaginalis was observed by scanning electron microscopy (SEM). The results demonstrated ET entangling T. vaginalis. Then, the colocalization of histone (H3) and myeloperoxidase (MPO) with DNA was observed via fluorescence confocal microscopy. Colocalization revealed the classic characteristics of DNA decorated with H3 and MPO. T. vaginalis significantly increased reactive oxygen species (ROS) and THP-1-derived ET. In addition, we measured the levels of lactic dehydrogenase (LDH) and the phosphorylation of the P38 and ERK1/2 MAPK signaling pathways. The results indicated that the formation of ET induced by T. vaginalis was related to phosphorylation of the P38 and ERK1/2 MAPK signaling pathways but not to LDH levels. These data confirmed the phenomenon of THP-1-derived ET being triggered by T. vaginalis in vitro; this process may play a pivotal role in innate immunity during defense against T. vaginalis infection.

Differential Protein Expressions in Virus-Infected and Uninfected Trichomonas vaginalis.

Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected (V+) and uninfected (V-) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in V+ compared with V- isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in V+ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in V+ and V- isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.

Seroprevalence of Dirofilaria immitis in Cats from Liaoning Province, Northeastern China

The present study was performed to investigate the seroprevalence and risk factors for Dirofilaria immitis infection in cats from Liaoning province, northeastern China. From October 2014 to September 2016, sera of 651 cats, including 364 domestic cats and 287 feral cats (332 females and 319 males) were assessed. They were tested for the presence of D. immitis antigen using SNAP Heartworm RT test kit. In this population, the average prevalence was 4.5%. Age and rearing conditions (feral or domestic) were found to be associated with the prevalence of D. immitis. The prevalence was significantly higher in feral cats compared with domestic cats (8.4% vs 1.4%, P 0.05), but older cats (≥3 years old) showed a statistically higher prevalence compared with younger cats ( 0.05), all these results suggest that outdoor exposure time may be one of the most important factors for D. immitis prevalence in cats. Results reveal that D. immitis are prevalence in domestic and feral cats in northeastern China, which indicates that appropriate preventive measures should be taken to decrease the incidence of feline heartworm disease in Liaoning province, northeastern China.

Codon Usage Bias and Determining Forces in Taenia solium Genome

The tapeworm Taenia solium is an important human zoonotic parasite that causes great economic loss and also endangers public health. At present, an effective vaccine that will prevent infection and chemotherapy without any side effect remains to be developed. In this study, codon usage patterns in the T. solium genome were examined through 8,484 protein-coding genes. Neutrality analysis showed that T. solium had a narrow GC distribution, and a significant correlation was observed between GC12 and GC3. Examination of an NC (ENC vs GC3s)-plot showed a few genes on or close to the expected curve, but the majority of points with low-ENC (the effective number of codons) values were detected below the expected curve, suggesting that mutational bias plays a major role in shaping codon usage. The Parity Rule 2 plot (PR2) analysis showed that GC and AT were not used proportionally. We also identified 26 optimal codons in the T. solium genome, all of which ended with either a G or C residue. These optimal codons in the T. solium genome are likely consistent with tRNAs that are highly expressed in the cell, suggesting that mutational and translational selection forces are probably driving factors of codon usage bias in the T. solium genome.

A Novel Recombinant BCG Vaccine Encoding Eimeria tenella Rhomboid and Chicken IL-2 Induces Protective Immunity Against Coccidiosis

A novel recombinant Bacille Calmette-Guerin (rBCG) vaccine co-expressed Eimeria tenella rhomboid and cytokine chicken IL-2 (chIL-2) was constructed, and its efficacy against E. tenella challenge was observed. The rhomboid gene of E. tenella and chIL-2 gene were subcloned into integrative expression vector pMV361, producing vaccines rBCG pMV361-rho and pMV361-rho-IL2. Animal experiment via intranasal and subcutaneous route in chickens was carried out to evaluate the immune efficacy of the vaccines. The results indicated that these rBCG vaccines could obviously alleviate cacal lesions and oocyst output. Intranasal immunization with pMV361-rho and pMV361-rho-IL2 elicited better protective immunity against E. tenella than subcutaneous immunization. Splenocytes from chickens immunized with either rBCG pMV361-rho and pMV361-rho-IL2 had increased CD4+ and CD8+ cell production. Our data indicate recombinant BCG is able to impart partial protection against E. tenella challenge and co-expression of cytokine with antigen was an effective strategy to improve vaccine immunity.

Changes of cytokines in the Cricetulus barabensis and their albino mutant infected with trichinella spiralis / 中国比较医学杂志

Objective To explore the differences in immune responses between Cricetulus barabensis and their albino mutant infected with Trichinella spiralis.Methods The physiological parameters of blood , expression levels of IL-2 protein and IL-6 gene in the spleen were analyzed.Results The level of immune cells and cytokines of Cricetulus barabensis was higher than that in the albino mutant .Conclusions Cricetulus barabensis is a suitable model animal for research on long-term latent infection such as infection with Trichinella spiralis.

Differential dissolved protein expression throughout the life cycle of Giardia lamblia.

Giardia lamblia (G. lamblia) has a simple life cycle that alternates between a cyst and a trophozoite, and this parasite is an important human and animal pathogen. To increase our understanding of the molecular basis of the G. lamblia encystment, we have analyzed the soluble proteins expressed by trophozoites and cysts extracted from feces by quantitative proteomic analysis. A total of 63 proteins were identified by isobaric tags for relative and absolute quantitation (iTRAQ) labeling, and were categorized as cytoskeletal proteins, a cell-cycle-specific kinase, metabolic enzymes and stress resistance proteins. Importantly, we demonstrated that the expression of seven proteins differed significantly between trophozoites and cysts. In cysts, the expression of three proteins (one variable surface protein (VSP), ornithine carbamoyltransferase (OTC), ß-tubulin) increased, whereas the expression of four proteins (14-3-3 protein, α-tubulin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), protein disulfide isomerase 2 (PDI-2)) decreased significantly when compared with the levels of these proteins in trophozoites. The mRNA expression patterns of four of these proteins (OTC, α-tubulin, GAPDH, VSP) were similar to the expression levels of the proteins. These seven proteins appear to play an important role in the completion of the life cycle of G. lamblia.

The effect of magnolol on the Toll-like receptor 4/nuclear factor κB signaling pathway in lipopolysaccharide-induced acute lung injury in mice.

Magnolol, a hydroxylated biphenyl compound isolated from Magnolia officinalis has been reported to have anti-inflammatory properties. The purpose of this study was to evaluate the effect of magnolol on acute lung injury induced by lipopolysaccharide in mice. Male BALB/c mice were pretreated with dexamethasone or magnolol 1 h before intranasal instillation of lipopolysaccharide (LPS). 7 h after LPS administration, the myeloperoxidase in lung tissues, lung wet/dry weight ratio and inflammatory cells in the bronchoalveolar lavage fluid were determined. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) in the bronchoalveolar lavage fluid were measured by enzyme-linked immunosorbent assay (ELISA). The extent of phosphorylation of nuclear factor of inhibitory kappa B alpha (IκB-α), nuclear factor kappa-B (NF-κB) p65 and the expression of Toll-like receptor-4 (TLR4) were detected by western blot. The results showed that magnolol markedly attenuated the histological alterations in the lung; reduced the number of total cells, neutrophils, and macrophages in the bronchoalveolar lavage fluid; decreased the wet/dry weight ratio of lungs in the bronchoalveolar lavage fluid; down-regulated the level of pro-inflammatory mediators, including TNF-α, IL-1ß and IL-6; inhibited the phosphorylation of IκB-α, NF-κB p65 and the expression of TLR4, caused by LPS. Taken together, our results suggest that anti-inflammatory effects of magnolol against the LPS-induced acute lung injury may be due to its ability of inhibition TLR4 mediated NF-κB signaling pathways. Magnolol may be a promising potential therapeutic reagent for acute lung injury treatment.

Stevioside suppressed inflammatory cytokine secretion by downregulation of NF-κB and MAPK signaling pathways in LPS-stimulated RAW264.7 cells.

Stevioside, a diterpene glycoside isolated from Stevia rebaudiana, has been reported to have anti-inflammatory properties. However, the underlying molecular mechanisms are not well understood. The objective of this study was to investigate the molecular mechanism of stevioside in modifying lipopolysaccharide (LPS)-induced signal pathways in RAW264.7 cells. RAW264.7 cells were stimulated with LPS in the presence or absence of stevioside. The expression of pro-inflammatory cytokines was determined by enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction. Nuclear factor-κB (NF-κB), inhibitory kappa B (IκBα) protein, p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) were determined by western blot. The results showed that stevioside dose-dependently inhibited the expression of tumor necrosis factor-α, interleukin-6, and interleukin-1ß in LPS-stimulated RAW264.7 cells. Western blot analysis showed that stevioside suppressed LPS-induced NF-κB activation, IκBa degradation, phosphorylation of ERK, JNK, and P38. Our results suggest that stevioside exerts an anti-inflammatory property by inhibiting the activation of NF-κB and mitogen-activated protein kinase signaling and the release of proinflammatory cytokines. These findings suggest that stevioside may be a therapeutic agent against inflammatory diseases.

A fine balance between CCNL1 and TIMP1 contributes to the development of breast cancer cells.

Cyclin L1 (CCNL1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP1) are candidate genes involved in several types of cancer. However, the expression of CCNL1 and the relationship between CCNL1 and TIMP1 in breast cancer cells is unknown. Using patients' breast cancer tissues, the expression of CCNL1 and TIMP1 was measured by cDNA microarray and further confirmed by real-time RT-PCR and western blotting. Overexpression or repression of CCNL1 and TIMP1, individually or together, was performed in breast cancer MDA-MB-231 cells by transient transformation methods to investigate their role in breast cancer cell growth. Simultaneously, mRNA and protein expression levels of CCNL1 and TIMP1 were also measured. CCNL1 and TIMP1 expression was significantly elevated in breast cancer tissues compared with that in peri-breast cancer tissues of patients by cDNA microarray and these results were further confirmed by real-time RT-PCR and western blotting. Interestingly, in vitro experiments showed a stimulatory effect of TIMP1 and an inhibitory effect of CCNL1 on growth of MDA-MB-231 cells. Co-expression or co-repression of these two genes did not affect cell growth. Overexpression of CCNL1 and TIMP1 individually induced overexpression of each other. These data demonstrate that there is a fine balance between CCNL1 and TIMP1, which may contribute to breast cancer development.

IL-18 augments protective immunity of Sj23 plasmid DNA vaccine against Schistosoma japonicum in mice / 中国兽医学报

Two recombinant plasmids pVAX/Sj23 and pVAX/mIL-18 containing Schistosoma japonicum 23 000 membrane protein (Sj23) and murine IL-18 were evaluated for their ability to induce immune responses and to protect against S. japonicum challenge in mice. All animals vaccinated with pVAX/Sj23 alone or plus pVAX/mIL-18 developed specific anti-SWAP (soluble worm antigen preparation) ELISA antibody and splenocyte proliferation response,and co-injection of pVAX/mIL-18 significantly increased the production of IFN-γ and IL-2 compared with pVAX/Sj23 alone, indicating that IL-18 enhances the Th1-dominant immune response. The challenge experiment showed that worm reduction rates in pVAX/Sj23 group compared with control group (pVAX1) was 26.5% and in the pVAX/Sj23 plus pVAX/mIL-18 group was 41.9% ,and the hepatic egg reduction rates were 42.7 and 49.6%,respectively. These results indicated that co-injection of an IL-18 plasmid with Sj23 DNA vaccine efficiently improves the protective effect against S. japonicum infection.

Expressing of bovine viral diarrhea-mucosal disease virus Changchun184 strain of E2 gene in BCG / 中国兽医学报

E2 gene of BVDV Changchun 184 strain was cloned and inserted into the shuttle expression plasmid vector pMV261,the recombinant shuttle plasmid pMV261-E2 was constructed.Then pMV261-E2 was transformed into BCG successfully and obtained recombinant BCG which was resistive to kanamyein.The recombinant BCG were identifieated by PCR.E2 gene expression in recombinant BCG was induced in 45℃,then the SDS-PAGE and western blotting was used to analyze the expression product.The results indicated the BVDV E2 gene was expressd in BCG successfully.

The therapeutic effect of Nitazoxanide on the dogs infected with Giardia canis / 中国兽医学报

To observe the therapeutic effect of Nitazoxanide(NTZ) on dogs infected with Giardia canis trophozoites.Eight dogs were infected with Giardia canis trophozoites and divided into four groups rondomly,G1:2 dogs treated with Nitazoxanide at a single dose of 1 mg/kg body weight;G2:2 dogs treated with NTZ at a single dose of 2 mg/kg;G3:2 dogs treated with NTZ at a single dose of 41 mg/kg;G4:2 dogs treated without drugs as control.All groups were examined for Giardia canis cysts by Zinc Sulfate Flotation.Each group was subjected to collect stool per day and counted cysts.The results of G2 and G3 were negative after 1th day.G1 were negative after 4th days.The results indicated that NTZ at a dose of 2 mg/kg and 4 mg/kg in dogs had a faourable effect on the dogs infected with Giardia canis.

The changes of ?_1-adrenoceptor in ischeamic rabbit myocardium / 中国病理生理杂志

We used (H~3)-prazosin on rabbits to determine the changes of ?_1-adrenoceptor number (Bmax) and affinity (k_D) before and after ischemia, Within 15min of ischemia, we demonstrated that Bmax increased to twfold in ischeamic regions. The increase persisted for 30 min of ischemla and then began to fall at 45 min of ischemia. At the same time, in the nonischemic regions Bmax did not change. The results also showed that ischemia did not alter K_D markedly in both nonischeamic and ischeamic regions.

Construction of a eukaryotic expressing vector containing CP15 gene of C.parvum and its expression in Hela cells / 中国免疫学杂志

Objective:To construct an eukaryotic expressing vector pCR3 1 15 containing CP15 gene of Crypstosporidium parvum(C.parvum) and express it in Hela cells Methods:CP15 gene of C parvum was obtained from pMD18 T 15 disgested by BglⅡ and was inserted into eukaryotic expressing vector pCR3 1(+) in BamHⅠ site,and then Hela cells were transfected with recombinant by liposomes The transcription and expressed products of CP15 in the transfected Hela cells were assayed by RT PCR,ELISA and indiret immunofluorescence assay after screening with G418 Results:It showed that pCR3 1 15 was constructed successfully CP15 gene was transcripted in transfectants and CP15 protein with obvious biological activity was highly expressed in Hela cells Conclusion:CP15 gene in recombinant vector is proved to be expressed in Hela cells and obvious biological activity of expression production in transfected cells was detected