Asian Pacific Journal of Tropical Medicine

513-519

Authors: Rajeev Nagill, Tejinder Kaur, Jyoti Joshi, Sukhbir Kaur

Objective To analyze the protective efficacy of recombinant 78 kDa antigen of Leishmania donovani in combination with two adjuvants, that is, cationic liposomes or MPL-A against visceral leishmaniasis in BALB/c mice. Methods The genomic DNA of promastigotes was isolated and 583 bp of T cell epitopes of gene encoding 78 kDa was amplified using specific primers. The amplified gene was cloned into pET28c, transformed into Escherichia coli BL21 (DE3) and got expressed after IPTG induction. The recombinant protein was then purified using Ni-NTA and named r78. Three groups of mice were immunized with 10 μg of r78 plus MPL-A, r78 encapsulated in positively charged liposomes and control animals immunized with PBS. Two booster doses were given with the respective vaccine at an interval of 2 weeks each. Mice were challenged with 1 × 107Leishmania promastigotes and sacrificed on different post infection/challenge days. Results Immunization with r78 along with MPL-A and liposome-encapsulated r78 brought a significant reduction in parasite load. In comparison to the infected controls, the parasite load declined by 96.2% in mice immunized with r78 plus MPL-A and 97.23% in animals immunized with liposome-encapsulated r78. The immunized animals also exhibited profound DTH response. The serum antibody responses increased from 15 to 90 days post infection/challenge. Immunized animals showed greater IgG2a levels and lesser IgG1 levels in comparison to the infected controls. The splenocytes from immunized mice were cultured, stimulated with r78 and analyzed for cytokine profile. The levels of IL-2 and IFN-γ were greater in immunized animals as compared to control mice. Conclusions The study proves that r78 in combination with suitable adjuvants is a potential vaccine candidate and may be instrumental in control of visceral leishmaniasis.

526-531

Authors: El Moataz Bellah El Naggar, Marta Chalupová, Gabriela Pražanová, Tomáš Parák, Emil Švajdlenka, Milan Žemlička, Pavel Suchý

Objectives To investigate the effects of perorally administered juice on tetrachloromethane (CCl4)-induced hepatotoxicity model in rats. Methods Male Wistar rats were tube-administrated silymarin, Ecballium juice at 0.2 mL/kg and 0.7 mL/kg daily for 3 consequent days, i.e., 3.28 μg and 11.48 μg of cucurbitacin B per kg of body weight respectively. On the third day, liver damage was induced by intraperitoneal application of CCl4. On the fourth day, abdominal cavity was macroscopically examined and liver samples were taken for histopathological and immunochemical evaluation. HPLC was used to determine the content of the active substance cucurbitacin B. Results The experiment revealed that 0.7 ml/kg juice concentration expressed the highest pro-apoptotic activity, but with prevailing negative effects. Compared with the lower concentration, there was an observable vasodilatation with consequent interstitial hemorrhages and a larger scope of inflammatory damage, which suppressed the hepatoprotective effect. In the 0.2 mL/kg concentration, there was a smaller pro-apoptotic activity but other parameters had better results, and the liver parenchyma damage was reversible. Conclusions No reactions confirming the potentially allergic effect on laboratory rats were observed; its hepatoprotective and anti-inflammatory effect was confirmed on a model of acute liver damage.

538-542

Authors: Jang-Gi Choi, Ji-Young Choi, Su-Hyun Mun, Ok-Hwa Kang, Preeti Bharaj, Dong-Won Shin, Myong-Soo Chong, Dong-Yeul Kwon

Objective To investigate the antibacterial activity of SHH extracted with either water or ethanol against methicillin-resistant Staphylococcus aureus (MRSA) and combinatory antimicrobial effect with ciprofloxacin (CIP) by time kill assay and checkerboard dilution test. Methods The antibacterial activity determined by broth dilution method indicated that the antibacterial activity of Sami-Hyanglyun-Hwan (SHH) water extract (SHHW) and SHH ethanol extract (SHHE) ranged from 250 to 2 000 μg/mL and 125 to 1 000 μg/mL against MRSA, respectively. Results In the checkerboard method, the combinations of SHHE with CIP had a partial synergistic or synergistic effect against MRSA. The time-kill curves showed that a combined SHHE and CIP treatment reduced the bacterial counts dramatically after 24 h. Conclusions The present study demonstrates the therapeutic ability of SHHE against MRSA infections.

549-554

Authors: Kefale Awoke, Endalkachew Nibret, Abaineh Munshea

Objective To determine the prevalence of toxoplasmosis and to assess the possible risk factors associated with the infection among pregnant women attending antenatal care center at Felege Hiwot Referral Hospital, Bahir Dar town, northwest Ethiopia. Methods A hospital based cross-sectional study was designed to determine the prevalence of toxoplasmosis among pregnant women. Three hundred eighty four serum samples were collected from November 2013 to January 2014. Data on socio-demographic and predisposing factors were collected from each study participant with simple random sampling technique. The serum samples were examined for anti-Toxoplasma gondii (T. gondii) antibodies using latex agglutination test. Results The overall seroprevalence of T. gondii among the pregnant women was 18.5%. All of T. gondii positive cases found to be positive only for IgG antibody. Significant association was observed between seroprevalence and presence of domestic cats [AOR = 2.85, 95% CI: 1.66–4.90, P = 0.000], consumption of raw or undercooked meat [AOR = 1.98, 95% CI: 1.15–2.43, P = 0.014] and history of abortion [AOR = 2.47, 95% CI: 1.40–4.34, P = 0.002]. No significant association was observed between seroprevalence and socio-demographic characters, gestational age, gravidity, consumption of raw vegetable, and blood transfusion. Conclusions The seroprevalence of toxoplasmosis among pregnant women in Bahir Dar town was relatively high. Presence of domestic cats at home and consumption of raw or undercooked meat were identified as main risk factors for T. gondii infection. Therefore, health education towards avoiding eating raw or undercooked meat and avoiding contact with cats are recommended for prevention of miscarriage or defects during pregnancy.

560-564

Authors: Bin Wang, Chang-Jiang Lei, Rong Wu, Lei Li, Chun-Mei Deng, Wen-Xia Chen, Fu-Rong Hu, Hao-Cheng Long, Ze-Zhang Tao, Cheng Zeng, Jian-Bin Huang, Chun-Zhou

Objective To study the effect of lipoteichoic acid (LTA) and 5-FU on the expression of caspase-3, EGFR, TGF-α proteins of tumor tissue of H22 cancer bearing mice and its anti-tumor mechanism. Methods A total of 40 SPF grade Kunming mice were selected to establish H22 liver cancer model, and then the mice were divided into 4 groups at random with ten mice in each group. Group A was given saline lavage treatment, Group B was treated with 5-FU by intraperitoneal injection, Group C was treated with LTA by lump body injection; Group D was treated with LTA by lump body injection and 5-FU by intraperitoneal injection. Two weeks after the treatment, the mice in each group were executed and the tumor tissue was stripping and weighted, and the tumor growth inhibition ratio was calculated. Then the tumor tissue was processed for conventional embedding, sectioned to observe the expression of caspase-3, EGFR, TGF-α by immunohistochemical staining method. Results The tumor inhibitory rate o f Group D was significantly higher than Groups B and C (P < 0.05); B, the tumor inhibitory rate o f Group B had no statistical difference compared with Group C (P > 0.05). The IDO values of TGF-α, EGFR proteins in Groups B, C, D mice tumor tissue were significantly lower than that in group A (P < 0.05); while IDO value of caspase-3 in Groups B, C, D group mice tumor tissue was significantly higher than that in Group A (P < 0.05). The IDO value of TGF-α, EGFR in Group D mice tumor tissue were significantly lower than that in Groups B and C; While IDO value of aspase-3 in Group D was significantly higher than that in Groups B and C (P < 0.05). Conclusions LTA combined with 5-FU can effectively inhibit the tumorigenesis of H22 tumor bearing mice, increase the caspase-3 protein expression, inhibit TGF-α and EGFR protein expression, further promote tumor cell apoptosis and play a synergistic antitumor effect.

569-573

Authors: Qian-He Han, Zhong-Jie Shan, Jian-Ting Hu, Nan Zhang, Xue-Pei Zhang

Objective To investigate the relationship between genetic factor and prostate cancer (Pca) risk and the possible cause in it. Methods The polymorphisms of cytochrome P450 family 17 (CYPl7) rs743572, p27 V109G and androgen receptor (AR) gene CAG repeat length in peripheral blood from 70 cases and 70 controls were detected through the polymerase chain reaction-restriction fragment length polymorphism technique or short tandem repeat-polymerase chain reaction technique. Then, according to the results of case–control study, the recombinant plasmids containing the wild/mutant p27 gene were constructed and transfected Pca LNcap cells. After 24 and 72 h of transfection, the cell proliferative activity was determined by MTT method, cell cycle distribution and apoptosis was detected by flow cytometry, and the expression level of bcl-2, caspase-3 and p27 protein was determined by Western-blot. Results In three target polymorphisms, only p27 V109G polymorphism was related to Pca risk (P = 0.030, OR = 0.202, 95% CI = 0.042–0.973). Pca risk of p27-109G allele was lower than −109V allele (P = 0.006, OR = 0.285, 95% CI = 0.110–0.737). Cells transfected with wild/mutant p27 gene both showed the higher cells apoptosis rate and the lower cell proliferative activity than mock cells (P < 0.05 or 0.01), the regulatory effect of mutant p27 on cell proliferation and apoptosis was stronger than the wild p27 (P < 0.05). Conclusions p27-109G allele that could cause higher p27 protein expression than −109V allele in LNcap cells, maybe is the protective factor of Pca.

578-582

Authors: Hu Qu, Bo Ma, Hao-Feng Yuan, Zhong-Yang Wang, Sheng-Jie Guo, Jing Zhang

Objective To explore the effect of salinomycin on the metastasis and invasion of bladder cancer cell line T24 by regulating the related protein expression in the process of epithelial–mesenchymal transition (EMT), and to provide experimental basis for the treatment of urological tumors. Methods The bladder cancer cell line T24 was cultured in vitro. The rat bladder tumor model was established in vivo. The rats were randomized into two groups, among which the rats in the experiment group were given intraperitoneal injection of salinomycin, while the rats in the control group were given intraperitoneal injection of normal saline. The change of tumor cells in the two groups was observed. Transwell was used to detect the cell migration and invasion abilities, Real-time PCR was used to detect the expression of mRNA, while Western-blot was utilized for the determination of the expressions of E-cadherin and vimentin proteins. Results The metastasis and invasion abilities of serum bladder cancer cell line T24 after salinomycin treatment in the experiment group were significantly reduced when compared with those in the control group, and the tumor metastasis lesions were decreased from an average of 1.59 to 0.6 (P < 0.05). T24 cell proliferation in the experiment group was gradually decreasing. T24 cell proliferation at 48 h was significantly lower than that at 12 h and 24 h (P < 0.05). T24 cell proliferation at 24 h was significantly lower than that at 12 h (P < 0.05). T24 cell proliferation at each timing point in the experiment group was significantly lower than that in the control group (P < 0.05). The serum mRNA level and E-cadherin expression in the tumor tissues in the experiment group were significantly higher than those in the control group, while vimentin expression level was significantly lower than that in the control group (P < 0.05). Conclusions Salinomycin can suppress the metastasis and invasion of bladder cancer cells, of which the mechanism is probably associated with the inhibition of EMT of tumor cells.

590-593

Authors: Bao Shan, Jun-hong Cai, Shu-Ying Yang, Zhuo-Ri Li

Objective To study the relevant risk factors of polycystic ovarian syndrome (PCOS) of Li People so as to provide basis for early diagnosis and treatment of PCOS. Methods With case–control study method, 285 cases of PCOS of Li People were as recruited case group, and 580 cases of non-PCOS of female Li People as control group. Questionnaire was adopted to collect data regarding risk factors of PCOS, then the risk factors of PCOS was searched by univariate and multivariate analysis. Results Multivariate analysis showed that the risk factors of PCOS included in menstrual cycle disorder (OR = 5.824), bad mood (OR = 2.852), family history of diabetes (OR = 7.008), family history of infertility (OR = 11.953), menstrual irregularity of mother (OR = 2.557) and lack of physical exercise (OR = 1.866). Conclusions To target the high risk factors of menstrual cycle disorder, family history of diabetes, family history of infertility, family history of diabetes, bad mood and lack of physical exercise of female population, we should implement early screen, diagnose and treatment of POCS in order to reduce the incidence rate of PCOS and improve prognosis of PCOS.