Asian Pacific Journal of Tropical Medicine

914-918

Authors: Wiriyaporn Sumsakul, Wanna Chaijaroenkul, Kesara Na-Bangchang

Objective To investigate the propensity of plumbagin to inhibit the three isoforms of human cytochrome P450 (CYP), i.e., CYP1A2, CYP2C19, and CYP3A4 using human liver microsomes in vitro. Methods Inhibitory effects of plumbagin on the three human CYP isoforms were investigated using pooled human liver microsomes. Phenacetin O-deethylation, omeprazole hydroxylation and nifedipine oxidation were used as selective substrates for CYP1A2, CYP2C19 and CYP3A4 activities, respectively. Concentrations of paracetamol, 5-hydroxyomeprazole, and oxidized nifedipine were determined in microsomal incubation mixture using high-performance liquid chromatography. Results Plumbagin showed significant inhibitory effects on all CYP isoforms, but with the most potent activity on CYP2C19-mediated omeprazole hydroxylation. The IC50 (concentration that inhibits enzyme activity by 50%) values of plumbagin and nootkatone (selective inhibitor) for CYP2C19 were (0.78 ± 0.01) and (27.31 ± 0.66) μM, respectively. The inhibitory activities on CYP1A2-mediated phenacetin O-deethylation and CYP3A4-mediated nifedipine oxidation were moderate. The IC50 values of plumbagin and α-naphthoflavone (selective inhibitor) for CYP1A2 were (1.39 ± 0.01) and (0.02 ± 0.36) μM, respectively. The corresponding IC50 values of plumbagin and ketoconazole (selective inhibitor) for CYP3A4 were (2.37 ± 0.10) and (0.18 ± 0.06) μM, respectively. Conclusions Clinical relevance of the interference of human drug metabolizing enzymes should be aware of for further development scheme of plumbagin as antimalarial drug when used in combination with other antimalarial drugs which are metabolized by these CYP isoforms.

989-998

Authors: Uruj Tahir, Umair Hassan Khan, Muhammad Saad Zubair, Bahar-e-Mustafa

Dengue virus syndrome is an emerging global health challenge which is endemic in tropical countries like Pakistan. In recent years dengue incidences have increased considerably in different areas of Pakistan with more sever impacts on urban and peri-urban populations. This review is an effort to highlight the changing epidemiology of dengue fever, role of Government of Pakistan in disease management and control using preventive and community based approaches in the region. Moreover, there is an emphasis on application of Wolbachia as novel, inexpensive and environmentally benign candidate for control and eradication of dengue transmitting vectors.

1006-1012

Authors: Ke-Xin Yu, Ching-Lee Wong, Rohani Ahmad, Ibrahim Jantan

Objective To investigate the larvicidal activity, inhibition effect on development, histopathological alteration and morphological aberration induced by the extracts derived from seaweeds Bryopsis pennata (B. pennata), Sargassum binderi (S. binderi) and Padina australis in Aedes aegypti (Ae. aegypti) larvae and to characterize the phytochemical components of the three seaweeds. Methods Larvicidal activity of the seaweeds towards the larvae of Ae. aegypti was determined according to WHO. The inhibition effect of seaweeds was assessed by determining the mortality, adult emergence rate, larval and pupa duration of the treated larvae. Histopathological effect on midgut epithelium of larvae and morphological aberration induced by the methanol extracts were examined. Phytochemical analysis was done to determine the presence of alkaloids, saponins, steroids and terpenoids in the seaweeds. Results Chloroform partition of B. pennata extract exhibited the strongest larvicidal activity (LC50 = 82.55 μg/mL), followed by methanol extract of B. pennata (LC50 = 160.07 μg/mL) and chloroform partition of S. binderi extract (LC50 = 192.43 μg/mL). The methanol extract of S. binderi exhibited the strongest effect on prolongation of larval period (1.5-fold longer as compared to control) and resulted in strongest inhibition effect in adult emergence (98.67%). The histopathological study showed that larvae treated with seaweed extracts had cytopathological alteration of the midgut epithelium. The morphological observation revealed that the anal papillae and terminal spiracles of larvae were the common sites of aberrations. Conclusions The study provided information on various effects of seaweed extracts on Ae. aegypti. Further investigation on identifying the active compounds and their mechanisms of action is recommended.

1022-1026

Authors: Devika Iddawela, Kiruthiha Ehambaram, Susiji Wickramasinghe

Objective To identify worms obtained from patients with eye lesions and to describe the demographic factors of patients with ocular dirofilariasis. Methods A retrospective descriptive study was conducted in 31 worm samples from 30 patients referred by consultant ophthalmologists between 2006 and February 2014. Data on age, sex and site of the lesion were ascertained from the details given in the referral letters. Morphological identification of the worm was based on the maximum width, length and appearance of the cuticle. The sex of the worm was determined by the width, length and presence or absence of vulva opening. PCR was performed using Dirofilaria repens specific primers to confirm the species of worms which couldnot be identified morphologically. Results Most of the patients belonged to the age group of 40–49 years (mean age = 42 years). Majority of them were females (70%). Subconjunctival lesions were the most frequent presentation, while the rest (n = 4) were found on eyelids. Female worms were extracted from 18 cases, and 11 had male worms. One individual had both male and female worms in a single nodule. Adults were the most commonly affected. This pattern was different from the previous studies in Sri Lanka where the most common age group affected was younger than 9 years old. Conclusions The present study showed a considerably high incidence of ocular dirofilariasis, stressing the importance of implementing preventive measures to reduce the transmission of this zoonotic filarial disease.

1033-1037

Authors: Hong-Fen Ge, Guan-Zhi Chen, Min Pan, Hui Guo

Objective To observe the effect of the cyclophosphamide (CTX) on systemic lupus erythematosus mice by fluorescent quantitative polymerase chain reaction. Methods (BALB/c × C57BL/J6) F1 hybrid mice were used as recipients of donor lymphocytes which were injected intravenously. Mice were randomly divided into control group and treatment group, serum autoantibody and urinary protein were measured, renal pathological changes and the situation of fungal infection were observed. Results The reduced urinary protein, ameliorated renal lesions were found in the CTX-treated mice. Candida albicans and Aspergillus flavus were detected respectively in respiratory tract and gastrointestinal tract. Conclusions CTX can alleviate lupus nephritis and the fluorescent quantitative PCR is a rapid and sensitive method for the detection of fungal infection.

1043-1048

Authors: Jing-Bo Ji, Xue-Fu Li, Lei Liu, Guang-Zhong Wang, Xin-Feng Yan

Objective To study the effect of low intensity pulsed ultrasound (LIPUS) on the expression of tissue inhibitor of metalloproteinase-2 (TIMP-2) in the serum and expression of matrix metallopeptidase 13 (MMP-13) in the articular cartilage cells of rabbits with knee osteoarthritis (OA). Methods Inner patellar ligament defect method was used to establish the model of knee OA. Four weeks after the modeling, the arterial blood was drawn from the ear of each rabbit, while ELISA was employed to detect the expression of TIMP-2 in the serum. The chondrocytes were separated from animals in each group and then cultured in vitro. All rabbits were divided into control group, OA model group and OA + LIPUS group. Cells in the control and OA groups were not treated, while cells in the OA + LIPUS group were treated with LIPUS (40 mW/cm2, 1 time/day). Cells were collected 7 d later and the RNA and total protein were extracted respectively. Real-time PCR and Western blotting were employed to analyze the expression of MMP-13 in chondrocytes at the mRNA and protein level, respectively. Results The success rate of establishment of OA model was 83%. The results of ELISA showed that the content of TIMP-2 in the serum of animals with OA was 22.3%, lower than the one in the control group (P < 0.05). Compared with the normal control group, the expression of TIMP-2 in the OA model group was significantly increased, while the expression of MMP-13 was significantly increased (P < 0.05). After the stimulation of LIPUS, the expression of TIMP-2 and MMP-13 was close to the one in the normal control group. Conclusions The inner patellar ligament defect method is a mature method to establish the rabbit OA model, with high success rate. The expression of serum TIMP-2 in the OA model group is significantly decreased. LIPUS can up-regulate TIMP-2 and down-regulate MMP-13.

1055-1059

Authors: Guan-Li Zhang, Ling Jiang, Qian Yan, Rong-Hui Liu, Lu Zhang

Objective To observe the anti-tumor effect of matrine combined with cisplatin on U14 rat models of cervical cancer. Methods A total of 80 female Kunming rats were used to establish U14 rat models of cervical cancer and then divided into groups I, II, III, IV, with 20 rats in each. For Group I, the control group, injection of normal saline was given around the tumors. For Group II, injection of 2 mg/kg cisplatin was given around the tumors. For Group III, injection of 75 mg/kg matrine was given around the tumors while the combined injection of matrine and cisplatin was given for Group IV with the same doses as Groups II and III. The animals were sacrificed 10 d after the injection and tumors were taken out for the comparisons of tumor weights after injection and calculation of anti-tumor rates, while thymus and spleen were taken for thymus index and spleen index. Blood in eyeball was collected for determination of changes in serum creatinine and urea nitrogen levels. Sections of tumor issue were prepared and morphological changes in tumor tissue cells were observed by using immunohistochemistry technique. Results After injection, the thymus index and spleen index in Groups III and IV were significantly higher than those in Groups I and II (P < 0.05) while the two indexes in Group II were significantly lower than Group I (P < 0.05). The tumor weights in Groups II and IV were significantly smaller than those in Groups I and III (P < 0.05) with significantly higher anti-tumor rates than Groups I and III (P < 0.05). The serum creatinine and urea nitrogen levels in Groups III and IV were significantly lower than Group II (P < 0.05) and the two indicators in Group III were significantly lower than those in Group IV (P < 0.05). The observation under the histological microscope showed densely arranged tumor cells in Group I, growing as a crumby structure and diffuse appearance, with hyperchromatic and large nuclei, and abundant cytoplasm. In the case of Group II, it showed less tumor cells, with extensive degenerative necrosis, sparse arrangement and karyopyknosis as well as karyoclasis. For Group III, necrosis of tumor cells in different sizes and heterogeneous color in nuclei were observed. For Group IV, the number of tumor cells was significantly smaller than Groups I and III and the tumor cells presented an appearance of crumby structure as cancer nests, with more proliferation of connective tissue. Conclusions The treatment of matrine combined with cisplatin can significantly improve the anti-tumor effect on U14 rats with cervical cancer, which can be a new option for the treatment for cervical cancer.

1064-1070

Authors: Yong Yang, Da-Peng Li, Na Shen, Xiao-Cheng Yu, Jie-Bao Li, Qi Song, Jia-Heng Zhang

Objective To investigate the expression of targeting protein for Xenopus kinesin-like protein 2 (TPX2) in breast cancer tissue and to explore its role in proliferation, migration and invasion of breast cancer cells. Methods The mRNA and protein expressions of TPX2 in breast cancer tissue and cell lines were assessed by quantitative RT-PCR and Western blot. The effect of TPX2 with RNA interference on proliferation, invasion and migration of breast cancer cells was observed by MTT and Transwell assays. Results Both mRNA and protein expressions of TPX2 were upregulated in breast cancer tissues compared to tumor-adjacent tissue. TPX2 expression was also upregulated in breast cancer cell lines, and the TPX2 interfered by small interfering RNA could inhibit the proliferation, invasion and migration of breast cancer cells by inhibiting matrix metalloproteinase-2 and matrix metalloproteinase-9. Conclusions Significantly upregulated TPX2 expression is observed in breast cancer tissue and cells, and contributes to promote the proliferation, migration and invasion of breast cancer cells.

1076-1078

Authors: Wen-Ning Li, Jia-Li Wei, Ming Wu, Wei Wu, Yun Huang, Mao-Wei Xie, Hui Han

Objective To prove whether astrocyte elevated gene-1 (AEG-1) plays a role in high glucose-stimulated Rho kinase activation and epithelial–mesenchymal transition (EMT) in human renal tubular epithelial (HK-2) cells. Methods The protein levels of AEG-1, alpha-smooth muscle actin, E-cadherin and MYPT1 were determined by Western blot. Results AEG-1 protein level was upregulated in HK-2 cells stimulated with high glucose. AEG-1 siRNA downregulated Rho kinase protein expression and blocked high glucose-induced EMT. Conclusions Our results show that AEG-1 acts a key role in high glucose-induced activation of Rho kinase and EMT in HK-2 cells.