Asian Pacific Journal of Tropical Medicine

176-184

Authors: Watcharee Attatippaholkun, Panyupa Pankhong, Ananda Nisalak, Siripen Kalayanarooj

Objective To study evolutionary relationship of the 5'untranslated regions (5'UTRs) in low passage dengue3 viruses (DEN3) isolated from hospitalized children with different clinical manifestations in Bangkok during 24 year–evolution (1977–2000) comparing to the DEN3 prototype (H87). Methods The 5'UTRs of these Thai DEN3 and the H87 prototype were amplified by RT–PCR and sequenced. Their multiple sequence alignments were done by Codon Code Aligner v 4.0.4 software and their RNA secondary structures were predicted by MFOLD software. Replication of five Thai DEN3 candidates comparing to the H87 prototype were done in human (HepG2) and the mosquito (C6/36) cell lines. Results Among these Thai DEN3, the completely identical sequences of their first 89 nucleotides, their high–order secondary structure of 5'UTRs and three SNPs including the predominant C90T, and two minor SNPs including A109G and A112G were found. The C90T of Thai DEN3, Bangkok isolates was shown predominantly before 1977. Five Thai DEN3 candidates with the predominant C90T were shown to replicate in human (HepG2) and the mosquito (C6/36) cell lines better than the H87 prototype. However, their highly conserved sequences as well as SNPs of the 5'UTR did not appear to correlate with their disease severity in human. Conclusions Our findings highlighted evolutionary relationship of the completely identical 89 nucleotide sequence, the high–order secondary structure and the predominant C90T of the 5'UTR of these Thai DEN3 during 24 year–evolution further suggesting to be their genetic markers and magic targets for future research on antiviral therapy as well as vaccine approaches of Thai DEN3.

191-196

Authors: Sung-Suk Suh, So Jung Kim, Jinik Hwang, Mirye Park, Taek-Kyun Lee, Eui-Joon Kil, Sukchan Lee

Objective To screen the fatty acid (FA) composition of 20 marine microalgae species, including seven Diophyceae, six Bacillariophyceae, four Chlorophyceae, two Haptophyceae and one Raphidophyceae species. Methods Microalgal cells cultured at the Korea Institute of Ocean Science & Technology were harvested during the late exponential growth phase and the FA composition analyzed. Results The FA composition of microalgae was species–specific. For example, seven different species of Dinophyceae were composed primarily of C14:0, C16:0, C18:0, C20:4n–6, C20:5n–3 and C22:6n–3, while C14:0, C16:0, C16:1, C18:0, C20:5n–3 and C22:6n–3 were abundant FAs in six species of Bacillariophyceae. In addition, four Chlorophyceae, two Haptophyceae and one Raphidophyceae species all contained a high degree of C16:1n–7 [(9.28–34.91)% and (34.48–35.04)%], C14:0 [(13.34–25.96)%] and [(26.69–28.24)%], and C16:0 [(5.89–29.15)%] and [(5.70–16.81)%]. Several factors contribute to the nutritional value of microalgae, including the polyunsaturated FA content and n–3 to n–6 FA ratio, which could be used to assess the nutritional quality of microalgae. Conclusions This study is the first comprehensive assessment of the FA composition and nutritional value of microalgae species in South Korea, and identifies the potential utility of FAs as species–specific biomarkers.

203-205

Authors: Zahra Meshkat, Sadegh Chinikar, MohammadTaghi Shakeri, Lida Manavifar, Maryam Moradi, Hessam Mirshahabi, Tahmineh Jalali, Sahar Khakifirouz, Nariman

Objective To evaluate the prevalence of West Nile virus seropositivity in the general population of Mashhad, Northeast of Iran. Methods One hundred and eighty two individuals living in the city of Mashhad were studied using cluster sampling method. Both IgM and IgG antibodies against WNV were detected by ELISA method. Results In this study, the overall IgG seroprevalence of positive West Nile virus was 11%; however, IgM antibody was not found in the participants. Conclusions Our study suggested that the prevalence rate of West virus is considerable in Mashhad city. It seems necessary for clinicians and health care workers to be aware of WNV infection in the Northeast Iran.

209-213

Authors: Hong-Juan Chang, Tian-Fa Li, Jun-Li Guo, You-Ling Lan, Yue-Qiong Kong, Xin Meng, Xian-Ji Ma, Xiao-Ling Lu, Wei-Ying Lu, Shao-Jiang Zheng

Objective To explore effect of high glucose on expression of osteoprotegerin (OPG) and receptor activator of NF– κ B ligand (RANKE) in rat aortic vascular smooth muscle cells. Methods SD rats were intraperitoneally injected with streptozotocin, OPG and RANKL expression in rat thoracic aortas were detected by immunohistochemical staining. In cultured vascular smooth muscle cells (VSMCs) (A7r5), qRT–PCR and Western blot analysis were used to examine the mRNA and protein levels of OPG and RANKL. Results Our results demonstrated that OPG expression was increased in hyperglycemic rat aortic VSMCs, while RANKL expression was decreased. Besides, in vitro experiments high glucose induced OPG expression, but depressed RANKL expression by dose– and time–dependent manner in cultured A7r5. Conclusions Our findings suggested that high glucose could promote the expression of OPG, and inhibit the expression of RANKL in VSMCs, which may be partly be the molecular mechanism of diabetic vascular calcification.

220-224

Authors: Ping Gao, Ya-Jie Gao, Hong-Lu Liang

Objective To investigate the effects of NF– κ B inhibitor pyrrolidine dithiocarbamate hydrochloride (PDTC) on vascular endothelial growth factor (VEGF) and endostatin expression in mice with Lewis lung cance; and its mechanism. Methods Mice survival rate and anti–tumor effects were observed in different concentrations of NF– κ B inhibitor PDTC after the Lewis lung cancer mice model was established. VEGF and endostatin expressions were detected by immunohistochemical assay. Results Lewis lung cancer was be inhibited by 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg of NF– κ B inhibitor PDTC (P<0.05). Microvessel density (MVD) in 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF– κ B inhibitor PDTC groups were significantly lower than the control group (P<0.05). Immunohistochemical assay results showed that VEGF and endostatin expressions in the 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF–κ B inhibitor PDTC groups were significantly lower than the control group (P<0.05). Western blot results also showed that NF– κ B inhibitor PDTC could inhibit VEGF and endostatin expressions in tumor tissues. Conclusions NF– κ B inhibitor PDTC can inhibit tumor formation and reduce tumor angiogenesis in mice with Lewis lung cancer; and its mechanism maybe associated to VEGF and endostatin down–regulation.

229-233

Authors: Bing Sun, Jin-Fa Jiang, Cui-Mei Zhao, Chao-Hui Hu

Objective To observe the effects antiarrhythmic peptide 10 (AAP10) aon acute ventricular arrhythmia and the phosphorylation state of ischemic myocardium connexin. Methods Acute total ischemia and partial ischemia models were established by ceasing perfusion and ligating the left anterior descending coronary artery in SD rats. The effects of AAP10 (1 mg/L) on the incidence rate of ischemia–induced ventricular arrhythmia were observed. The ischemic myocardium was sampled to detect total–Cx43 and NP–Cx43 by immunofluorescent staining and western blotting. the total–Cx43 expression was detected through image analysis system by semi–quantitative analysis. Results AAP10 could significantly decrease the incidence of ischemia–induced ventricular tachycardia and ventricular fibrillation. During ischemic stage, total ischemia (TI) and AAP10 total ischemia (ATI) groups were compared with partial ischemia (PI) and AAP10 partial ischemia (API) groups. The rates of incidence for arrhythmia in the ATI and API groups (10% and 0%) were lower than those in the TI and PI groups (60% and 45%). The difference between the two groups was statistically significant (P=0.019, P=0.020). The semi–quantitative analysis results of the ischemic myocardium showed that the total–Cx43 protein expression distribution areas for TI, ATI, PI and API groups were significantly decreased compared with the control group. On the other hand, the NP–Cx43 distribution areas of TI, ATI, PI and API groups were significantly increased compared with the control group (P>0.05). AAP10 could increase the total–Cx43 expression in the ischemic area and decrease the NP–Cx43 expression. Western blot results were consistent with the results of immunofluorescence staining. Conclusions AAP10 can significantly decrease the rate of incidence of acute ischemia–induced ventricular tachycardia and ventricular fibrillation. Acute ischemic ventricular arrhythmias may have a relationship with the decreased phosphorylation of Cx43 induced by ischemia. AAP10 may stimulate the phosphorylation of Cx43 by increasing the total–Cx43 expression and decreasing the NP–Cx43 expression in the ischemic area, so as to decrease ventricular arrhythmia.

239-242

Authors: Xun Yang, Bei-Bei Chen, Ming-Hua Zhang, Xin-Rong Wang

Objective To study the role of microRNA–126 in the development of lung cancer. Methods The biological function of microRNA–126 was detected using EdU assay and CCK–8 assay; the target gene of microRNA–126 was analyzed using real time RT–PCR and Western blot assay. Results In A549 cell line, overexpression of microRNA–126 inhibits the proliferation rate; VEGF is the target gene of microRNA–126; microRNA–126 exerts its function via regulating VEGF protein level. Conclusions microRNA–126 inhibits the proliferation in A549 cell line.

249-252

Authors: Cheng-De Wang, Xin-Rong Wang, Chao-Yang Wang, Yi-Jun Tang, Ming-Wen Hao

Objective To study the relevance of EGFR gene mutation with pathological features and prognosis in patients with non–small–cell lung carcinoma. Methods A total of 297 patients from July 2009 to May 2013 were chosen as objects. EGFR gene mutation were detected with fluorescence quantitative PCR. Relevance of EGFR gene mutation with clinical and pathological features was analyzed, and the prognosis of EGFR– mutant–patients and that of EGFR– wide type–patients was compared. Results In 297 patients, 136 (45.79%) showed EGFR gene mutation. EGFR gene mutation had no significant relevance with age, gender, smoking history, family history of cancer and clinical stage (P>0.05); there was significant relevance between EGFR gene mutation and blood type, pathologic types, differentiation and diameter of cancer (P<0.05). The difference between prognosis of EGFR– mutant–patients and that of EGFR– wide type–patients was statistical significance (P<0.05). Conclusions EGFR gene mutation has significant relevance with pathological features, the prognosis of EGFR–mutant–patients is better than that of EGFR– wide type–patients.