Asian Pacific Journal of Tropical Medicine

529-538

Authors: Chayan Kumar Saha, Md. Mahbub Hasan, Md. Saddam Hossain, Md. Asraful Jahan, Abul Kalam Azad

Objective To explore a common B- and T-cell epitope-based vaccine that can elicit an immune response against encephalitis causing genus Henipaviruses, Hendra virus (HeV) and Nipah virus (NiV). Methods Membrane proteins F, G and M of HeV and NiV were retrieved from the protein database and subjected to different bioinformatics tools to predict antigenic B-cell epitopes. Best B-cell epitopes were then analyzed to predict their T-cell antigenic potentiality. Antigenic B- and T-cell epitopes that shared maximum identity with HeV and NiV were selected. Stability of the selected epitopes was predicted. Finally, the selected epitopes were subjected to molecular docking simulation with HLA-DR to confirm their antigenic potentiality in silico. Results One epitope from G proteins, one from M proteins and none from F proteins were selected based on their antigenic potentiality. The epitope from the G proteins was stable whereas that from M was unstable. The M-epitope was made stable by adding flanking dipeptides. The 15-mer G-epitope (VDPLRVQWRNNSVIS) showed at least 66% identity with all NiV and HeV G protein sequences, while the 15-mer M-epitope (GKLEFRRNNAIAFKG) with the dipeptide flanking residues showed 73% identity with all NiV and HeV M protein sequences available in the database. Molecular docking simulation with most frequent MHC class-II (MHC II) and class-I (MHC I) molecules showed that these epitopes could bind within HLA binding grooves to elicit an immune response. Conclusions Data in our present study revealed the notion that the epitopes from G and M proteins might be the target for peptide-based subunit vaccine design against HeV and NiV. However, the biochemical analysis is necessary to experimentally validate the interaction of epitopes individually with the MHC molecules through elucidation of immunity induction.

544-548

Authors: Azar Shokri, Javad Akhtari, Masoud Keighobadi, Mahdi Fakhar, Saeed Hosseini Teshnizi, Saeed Emami, Sajede Sadjjadian

Objective To evaluate the effect of doxorubicin and its pegylated liposomal formulation (Doxil, Caelyx) on in vitro susceptibility of promastigote and amastigote stages of Leishmania major. Methods Throughout in vitro assays the IC50 was calculated in the promastigotes and amastigotes forms in J774 macrophage cell line. Also as cytotoxicity in J774 cell line macrophages. Results Doxorubicin and Doxil showed the same activity against promastigote form with IC50 values of 10.49 μg/mL and 9.63 μg/mL, respectively. Similarly, the amastigote stage was susceptible at concentration of at least 1 μg/mL when compared to positive control (P < 0.0001). Also, cytotoxicity assay against macrophage revealed no toxicity on the host cells at IC50 concentrations. Conclusions Our findings demonstrated the efficacy of both doxorubicin and its pegylated liposomal formulation on L. major at low concentrations. Further researches are needed for evaluating the safety of drugs in animal model particularly as topical formulation.

557-565

Authors: Ayat O.S. Montasser, Hanan Saleh, Omar A. Ahmed-Farid, Aida Saad, Mohamed-Assem S. Marie

Objective To compare the degree of ameliorative effects of Melatonin (MEL), Ursodeoxycholic acid (UDCA) and Balanites aegyptiaca (BA) against hepatotoxicity induced by MTX for one month. Methods Eighty adult male rats (Sprague Dawely) weighing (190 ± 10 g), were randomly divided into eight equal groups: Control, MTX, MEL, BA, UDCA, MTX + MEL, MTX + BA, MTX + UDCA. Liver function biomarker enzymes, liver tissue oxidative stress parameters, together with total antioxidant capacity and tumor necrosis factor (TNF-α) were determined. Histopathological and immunohistochemistry examinations for TNF-α were also done. Results MTX showed significant increase in alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), total and direct bilirubin, as well as TNF-α levels, oxidized glutathione (GSSG), malodialdehyde (MDA) and nitric oxide (NO). Whereas total protein, albumin, total antioxidant capacity, reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) levels were significantly decreased in MTX treated group. These alterations were improved by MEL and BA treatment, whereas no improvement was noticed in UDCA treatment. Conclusions BA may be as promising as MEL in the hepatoprotection against MTX toxicity through their antioxidant and radical scavenging activities. In addition, it is not recommended to co-administer UDCA with MTX as it enhanced inflammation and damage to the liver.

572-575

Authors: Zhai Lu, Hua Chen, Xiao-Mei Zheng, Man-Ling Chen

Objective To study the regulatory effect and molecular mechanism of juglone on apoptosis of cervical cancer Hela cells. Methods Cervical cancer Hela cells were cultured and treated with different dosages of juglone (10, 20, and 40 μmol/L, respectively) and c-Jun N-terminal kinase (JNK) inhibitor SP600125 (10, 20, and 40 μmol/L, respectively). Then cellular proliferative activity and the expression of JNK/c-Jun pathway molecule and apoptotic molecule in the cells were detected. Results After 6, 12, 18 and 24 h of treatment, the value for proliferative activity of cells treated with juglone was significantly lower than that of control group (P < 0.05), and the anti-proliferative effect was more significant as the treatment period and juglone dosage increased (P < 0.05). The protein expressions of Bax, CytC, Fas, FasL, Caspase-3, p-JNK and p-c-Jun in cells treated with juglone were significantly higher than those of control group (P < 0.05), and the protein expressions of Bax, CytC, Fas, FasL, Caspase-3, p-JNK and p-c-Jun increased more remarkably as the juglone dosage increased (P < 0.05). In cells treated with 40 μmol/L juglone and SP600125, the protein expressions of Bax, CytC, Fas, FasL and Caspase-3 were significantly lower than those of cells treated with 40 μmol/L juglone (P < 0.05), and the protein expressions of Bax, CytC, Fas, FasL and Caspase-3 reduced more remarkably as the SP600125 dosage increased (P < 0.05). Conclusion Juglone can increase the expression of apoptotic molecules in mitochondrial pathway and death receptor pathway by activating JNK/c-Jun pathway, thus inducing apoptosis of cervical cancer cells.

582-587

Authors: Xiao-Mei Tang, Zhi-Kai Liao, You-Wei Huang, Xi Lin, Liang-Cai Wu

Objective To investigate whether atractylenolide Ⅰ (ATL-Ⅰ) has protective effect on lipopolysaccharide (LPS)-induced disseminated intravascular coagulation (DIC) in vivo and in vitro, and explore whether NF-κB signaling pathway is involved in ATL-Ⅰ treatment. Methods New Zealand white rabbits were injected with LPS through marginal ear vein over a period of 6 h at a rate of 600 μg/kg (10 mL/h). Similarly, in the treatment groups, 1.0, 2.0, or 5.0 mg/kg ATL-Ⅰ were given. Both survival rate and organ function were tested, including the level of alanine aminotransferase (ALT), blood urine nitrogen (BUN), and TNF-α were examined by ELISA. Also hemostatic and fibrinolytic parameters in serum were measured. RAW 264.7 macrophage cells were administered with control, LPS, LPS + ATL-Ⅰ and ATL-Ⅰ alone, and TNF-α, phosphorylation (P)-IκBα, phosphorylation (P)-NF-κB (P65) and NF-κB (P65) were determined by Western blot. Results The administration of LPS resulted in 73.3% mortality rate, and the increase of serum TNF-α, BUN and ALT levels. When ATL-Ⅰ treatment significantly increased the survival rate of LPS-induced DIC model, also improved the function of blood coagulation. And protein analysis indicated that ATL-I remarkably protected liver and renal as decreasing TNF-α expression. In vitro, ATL-I obviously decreased LPS-induced TNF-α production and the expression of P-NF-κB (P65), with the decrease of P-IκBα. Conclusions ATL-Ⅰ has protective effect on LPS-induced DIC, which can elevate the survival rate, reduce organ damage, improve the function of blood coagulation and suppress TNF-α expression by inhibiting the activation of NF-κB signaling pathway.

594-598

Authors: Wen-Yong Xue, Jin-Chun Qi, Lei Du

Objective To analyze the invention effect of curcumin on chronic urinary tract infection in rats and explore its possible mechanism of action. Methods The experimental animals were randomly divided into three groups, normal, model and curcumin group. Chronic urinary tract infection models were built for model group and curcumin group by injecting coliform fluid into the cavity of bladder. From the first day of modeling, rats in the curcumin group were injected with 150 mg/kg curcumin, while rats in normal group and model group were given no other treatment. The treatment lasted for 14 d. The white blood cell counts in blood and urine, bacterial colony count in urine and renal tubular functional indexes of rats in all groups at day 1, 7, and 14 after treatment were detected. Urine β2-microglobulin (β2-MG), urinary N-acetyl-β-D glucosaminidase (NAG) levels were used to detected the inflammatory cytokines in serum after treatment including the contents of IL-6, IL-8, IL-10 and monocyte chemoattractant protein-1 (MCP-1), and real-time PCR was employed to determine the expression of mRNA of toll-like receptor 2 (TLR-2) and TLR-4 in renal tissues and bladder tissues of all groups after treatment. Results The white blood cell counts at day 1 and 7 after treatment in rats of model group and curcumin group were significantly higher than those of normal group at the same time points, while the white blood cell counts of the curcumin group were significantly lower than those of model group (P < 0.05). The urine white blood cell counts in rats of model group at day 1, 7 and 14 were all significantly higher than those of normal group at the same time points; those in the curcumin group were significantly lower than those of the model group at day 1, 7 and 14 at the same time points (P < 0.05). The bacterial colony counts of urine in rats of model group and curcumin group at day 1, 7 and 14 were all significantly higher than those of normal group at the same time points, while the counts of curcumin group were significantly lower than those of model group at the same time points (P < 0.05). Levels of urine β2-MG, NAG, IL-6, IL-8, IL-10, MCP-1 and expression of TLR2 mRNA and TLR4 mRNA in renal and bladder tissues in rats of model group were significantly higher than those of the normal group, while these variables of the curcumin group were significantly higher than those of the normal group but lower than those of model group (P < 0.05). Conclusions Curcumin can significantly improve the symptoms of chronic urinary tract infections, protect renal tubular function, and also decline inflammatory responses by influencing the expressions of TLR2 mRNA and TLR4 mRNA so as to exert its curative effect on chronic urinary tract infections.

607-611

Authors: Xiang-Ying Li, Jian-Qiang Chen, Yi-Kai Xu, Xiang-Jun Han

Objective To explore the magnetic resonance diffusion tensor imaging (MR-DTI) features of in the late stage of Wistar rat C6 brain glioma, and the relationship between fractional anisotropy value and tumor microarchitecture. Methods The concentration of more than 1.0 × 106/10 μL glioma cells and complete medium were injected stereotactically into the right caudate nucleus of the experimental group (n = 35) and control group (n = 10), respectively. Conventional MRI, DTI, and enhanced T1WI scans were Performed using the GE Signa HD × 3.0T MRI scanner about 3–4 weeks after implantation for the rats. Postprocessing was done using the DTI specific software Function Tool to gain FA image. Many ROIs were drawn avoiding hemorrhage, necrosis areas in tumor parenchyma, the value of FA was recorded. Each surviving rat brain was examined histologically using HE and immunohistochemical staining for VEGF and CD34. Pearson correlation analysis was used to determine the relationships between FA values and VEGF, MVD, cell density, respectively. Results A total of 35 tumor-bearing rats were confirmed the tumor formation by the subsequent MRI and pathological examination. The mean FA values of the tumor and the contralateral brain tissue were 0.17 ± 0.03 and 0.31 ± 0.05 respectively, and the difference was statistically significant (t = 12.80, P < 0.05). The mean FA value of grade III glioma (n = 12) was 0.16 ± 0.03, and the average FA value of grade IV glioma (n = 23) was about 0.18 ± 0.04. There was no significant difference between the two groups (t = 1.92, P > 0.05). FA value in the late stage of Wistar rat C6 brain glioma has significant positive correlation to VEGF, MVD, cell density. The correlation coefficients between FA and VEGF, MVD, and cell density were 0.67, 0.65 and 0.71 (P < 0.05), respectively. Conclusions The FA value of rat glioma tumor in the late stage can preoperatively provide an accurate, reliable and noninvasive imaging monitoring method to evaluate the microstructure of glioma (cell density, the extent of angiogenesis, fiber bundle integrity and tumor cell infiltration and so on), predict the biological behavior of the tumor and make out surgical

614-618

Authors: Yan Song, Jie-Ge Jiao, Ze-Jun Chen

In this article, based on the characteristics of tropical area, we analyze the beneficial and the disadvantage of tropical area to human health, points out the existing problems in elderly health management pattern in the tropics area, accordingly we discuss how to establish tropical characterized elderly health management, and put forward constructive suggestions.