Asian Pacific Journal of Tropical Medicine

420-425

Authors: Chang-Wei Hsieh, Wen-Ching Ko, Wai-Jane Ho, Chao-Kai Chang, Guo-Jhou Chen, Jen-Chieh Tsai

Objective To investigate suitable condition for extraction of the active components from Ajuga nipponensis (A. nipponensis). Methods Orthogonal experimental design was used to determine the optimal extraction parameters for ecdysterones and flavonoids. Finally, the hepatoprotective abilities of A. nipponensis extracts were evaluated by CCl4-induced animal models. Results Maximum yields of flavonoids (7.87 ± 0.10) mg/g and ecdysterones (0.73 ± 0.02) mg/g could be obtained when the extraction time was 50 min, the extraction temperature was 60 °C, and the ratio of sample to 70% (v/v) ethanol was 1:20 (w/w). The antioxidant property of A. nipponensis was correlated to the concentration of its extracts. At 5 mg/mL, A. nipponensis extract scavenged 84.8% of DPPH radical and had absorbance values of 2.43 ± 0.04 reducing power. Upon CCl4-induced liver injury, glutamic oxaloacetic transaminase and glutamic pyruvic transaminase decreased significantly after the mice were treated with A. nipponensis. Histological researches also explained that A. nipponensis reduced the extent of liver lesions induced by CCl4. Conclusions A. nipponensis exhibited potent antioxidant activity in chemical experimental models and hepatoprotective effect against CCl4-induced liver damage.

432-436

Authors: Silmara Baroni, Bruno Ambrosio da Rocha, Juliana Oliveira de Melo, Jurandir Fernando Comar, Silvana Martins Caparroz-Assef, Ciomar Aparecida Bersani-Amado

Objective To evaluate the effect of hydroethanolic extract of yacon on the hyperglycemia induced by streptozotocin (STZ) in neonatal rats. Methods Wistar rats aged two days old received an intraperitoneal injection of STZ (160 mg/kg); after seven weeks, glycosuria was determined and animals with glucose levels above 250 mg/dL were included in the study. Groups of diabetic and non-diabetic rats were treated orally with yacon extract at a dose of 400 mg/kg/d for 14 d. Tests were made for phytochemical characterization, glucose tolerance and toxicity. Results The results showed that treatment with the extract reduced the glucose levels of fed diabetic rats and did not change the glucose levels of fasting diabetic and normal rats. Additionally, also it was observed that treatment with the extract reduced blood glucose levels of diabetic rats during the oral and intravenous glucose tolerance tests. There was no change in body weight, liver enzymes or mortality with yacon extract treatment. The phytochemical screening revealed the presence of caffeic acid, chlorogenic acid, ferulic acid and gallic acid. Conclusions The data suggest that yacon extract reduces hyperglycemia, possibly by improving insulin sensibility through its phytochemicals constituents (phenolic compounds).

445-452

Authors: Marta Giovanetti, Teresa Milano, Luiz Carlos Alcantara, Laura Carcangiu, Eleonora Cella, Alessia Lai, Alessandra Lo Presti, Stefano Pascarella, Gianguglielmo Zehender, Silvia Angeletti, Massimo Ciccozzi

Objective To investigate the genetic diversity of Zika Virus (ZIKV) and the relationships existing among these circulating viruses worldwide. To evaluate the genetic polymorphisms harbored from ZIKV that can have an influence on the virus circulation. Methods Three different ZIKV dataset were built. The first dataset included 63 E gene sequences, the second one 22 NS3 sequences and the third dataset was composed of 108 NS5 gene sequences. Phylogenetic and selective pressure analysis was performed. The edited nucleic acid alignment from the Envelope dataset was used to generate a conceptual translation to the corresponding peptide sequences through UGene software. Results The phylogeographic reconstruction was able to discriminate unambiguously that the Brazilian strains are belonged to the Asian lineage. The structural analysis reveals instead the presence of the Ser residue in the Brazilian sequences (however already observed in other previously reported ZIKV infections) that could suggest the presence of a neutralization-resistant population of viruses. Conclusions Phylogenetic, evolutionary and selective pressure analysis contributed to improve the knowledge on the circulation of ZIKV.

460-464

Authors: Dechassa Tegegne, Mukarim Abdurahaman, Tadesse Mosissa, Moti Yohannes

Objectives To assess the seroprevalence and associated risk factors of toxoplasmosis among HIV patients in Agaro Town Health Center of Jimma zone. Methods Convenient sampling was used to collect blood samples from 135 patients attending anti-retroviral therapy from February to March 2015. Serum samples were tested for anti-Toxoplasma gondii (T. gondii) antibody by using latex agglutination test. Structured questionnaire was used to collect data on socio-demographic and risk factors associated with toxoplasmosis. Results Overall seroprevalence of toxoplasmosis was 80.7% (109/135, CI: 74.04–87.36). In multivariate analysis significant association was observed between anti-T. gondii seropositivity and raw meat consumption (OR: 3.514, CI: 1.167–10.581, P = 0.025), knowledge about toxoplasmosis (OR: 5.225, CI: 1.382, P = 0.015) and sex (OR: 4.023, CI: 1.382–19.762, P = 0.015). Conclusions Immuno compromised patients showed high rate of seropositivity and thus, it is highly advisable to introduce routine anti-T. gondii antibodies serological screening test prior to ART commencement.

470-473

Authors: Ying Kuang, Ying-Jie Nie

Objective To study the regulatory effects of miR-21 on breast cancer cell line proliferation and invasion as well as the downstream target genes. Methods Breast cancer cell lines MCF-7 were cultured and transfected with miR-21 mimics and the corresponding negative control mimics (NC mimics), and then MTS kits were used to detect cell viability. Transwell experiment was used to detect cell invasion ability, and fluorescence quantitative PCR was used to detect the expression of proliferation and invasion-related genes in cells. Results 24 h after transfection of miR-21 mimics and NC mimics, cell OD value and the number of invasive cells of miR-21 group were significantly higher than those of NC group, and mRNA contents of PDCD-4, FasL, PTEN, RhoB, Maspin, TIMP3 and RECK in cells were significantly lower than those of NC group. Conclusion miR-21 can promote the proliferation and invasion of breast cancer cell lines, and its downstream target genes include PDCD-4, FasL, PTEN, RhoB, Maspin, TIMP3 and RECK.

478-483

Authors: Xiao-Zhou Liu, Cheng-Jun Li, Su-Jia Wu, Xin Shi, Jian-Ning Zhao

Objective To investigate the role of survivin in osteosarcoma metastasis. Methods Small interfering RNA (siRNA) was used to knockdown the expression of survivin and α5 integrin in the human osteosarcoma cell line MG63. Western blotting and immunostaining methods were used to assessed the effect of survivin knockdown on the expression of α5 integrin through flow cytometry and fluorescence microscopy detection. Meanwhile, the invasion and migration of transfected cells in Transwell and wound healing assays were probed, and the growth situation of these cells transplanted into nude mice was monitored. Results Knockdown of survivin expression could inhibit the invasion and migration of osteosarcoma MG64 cells in vitro and the expression of α5 integrin on osteosarcoma MG64 cell surface, suggesting that survivin can inhibit the invasion and migration of osteosarcoma cells through downregulation of α5 integrin. Anti-α5 integrin antibody could also markedly decrease the capability of invasion and migration of osteosarcoma MG64 cells. Additionally, knockdown of survivin expression could slow the growth of osteosarcoma MG63 cells transplanted into nude mice. Conclusions Survivin-directed anti-tumor strategies might be an effective method in the treatment of osteosarcoma.

489-493

Authors: Zi-Shu Wang, Hong Jin, Dong-Ming Wang

Objective To observe the influence of different concentrations of homocysteine (Hcy) and hydrogen sulfide (H2S) on the secretion and activation of matrix metalloproteinase-2 (MMP-2) in cardiocytes so as to search for new ways to fight against myocardial tissue fibrosis. Methods Cardiocytes H9C2 was cultured in vitro and different concentrations of Hcy and H2S were added for 6-h and 24-h cultivation. MTT cell proliferation assay was applied to test the activation change of cardiocytes H9C2 after affecting by different concentrations of Hcy and H2S. ELISA and MTT were employed to detect the expression and enzymatic activity of MMP-2. Results The H9C2 cell inhibition of activity was more significant with 1 000 μmol/L of Hcy as compared with other concentrations (P < 0.001). With 2.5–100.0 μmol/L Hcy and 0.1, 1.0 and 10.0 mmol/L H2S, the activity of H9C2 did not change significantly (P > 0.05). Hcy with concentrations of 10, 50 and 100 μmol/L could increase the quantity of MMP-2 secreted by cardiocytes H9C2, and the interaction strength was concentration-dependent (P < 0.05). After interacting with 100 μmol/L of Hcy for 6 h, the zymogen activation effect of MMP-2 was stronger than that of the 2.5–25 μmol/L group (P < 0.05). After interacting with Hcy and H2S (1.0 mmol/L) for 6 h and 24 h, the activation effect of MMP-2 was stronger than those interacted with 10, 25, 50 and 100 μmol/L of Hcy (P < 0.05). Conclusions Hcy can increase the production of MMP-2 secreted by H9C2 cell and improve its zymogen activation. Besides, the interaction strength is concentration-dependent; while H2S can up-regulate the activation of MMP-2 and co-promote the activation of MMP-2 with Hcy as well.

499-502

Authors: Dong Guo, Ye-Wei Wang, Ji Ma, Lei Yan, Teng-Fei Li, Xin-Wei Han, Shao-Feng Shui

Objective To investigate the correlation between JNK signal and the apoptosis of VSMC as well as the expression of Cathepsin B and to explore the role of JNK signal in the development of cerebral aneurysm. Methods Rat models of cerebral aneurysm were established and histopathologic changes of cerebral aneurysm and the apoptosis of VSMC were analyzed. Rat models were respectively subject to subcutaneous injection of Cathepsin B siRNA and JNK inhibitor SP600125. Western blot technique was used to detect the expression of proteins like Cathepsin B, Caspase-3, and p-JNK. Spearman's rho was used to examine the correlation between p-JNK and Cathepsin B, as well as the expression of relevant proteins. Results The success rate of modeling rats with cerebral aneurysm was 88.75%. After the respective injection of Cathepsin B siRNA, SP600125 and their combination, the cell densities of VSMC of rats with cerebral aneurysm all increased significantly (P < 0.05 or P < 0.01), but the apoptosis rate of VSMC decreased significantly (P < 0.01). Compared with normal rats, the expression of Cathepsin B, Caspase-3 and p-JNK in Cerebral aneurysm models increased significantly. Effectively intervening Cathepsin B genes with Cathepsin B siRNA could significantly inhibit the expression of Cathepsin B and Caspase-3, but hardly influence the expression of p-JNK. JNK inhibitor SP600125 had no influence on the expression of Cathepsin B and Caspase-3, but effectively inhibited the expression of p-JNK. In cerebral aneurysm tissues, positive correlation was observed between the expression of p-JNK and Cathepsin B, the correlation coefficient was r = 0.640. Conclusion After the attack of cerebral aneurysm, proteins like Cathepsin B, Caspase-3 and p-JNK are all involved in the apoptosis of VSMCs. This process may be realized by Cathepsin B which activates the apoptosis mechanism of Caspase-3 and mediate the apoptosis of VSMC through the JNK signaling pathway. Therefore, silencing Cathepsin B gene or inhibiting the conduction through JNK signaling pathway can mitigate the apoptosis of vascular smooth muscle cells in cerebral aneurysm.

508-512

Authors: Ting-Ting Ji, Xuan Huang, Jie Jin, Sheng-Hua Pan, Xiao-Ju Zhuge

Objective To discuss the expression of long noncoding RNA TUG1 (lncRNA-TUG1) in gastric carcinoma (GC) and its effects on the transferring and invading capacity of gastric carcinoma cells. Methods Forty cases of carcinoma tissue and para-carcinoma tissue were selected from GC patients who underwent surgical removal in Zhejiang Provincial Hospital of Chinese Traditional Medicine and Wenzhou Central Hospital from January, 2013 to December, 2014; the expressing level of lncRNA-TUG1 in GC and para-C tissues was detected by applying the qRT-PCR technique. The correlation between lncRNA-TUG1 expression and patients' clinical data was classified and analyzed. SGC-7901 cells were transfected using lncRNA-TUG1 specific siRNA. Changes of the transferring and invading capacity of siRNA-transfected SGC-7901 cells were scratch-tested and transwell-detected. qRT-PCR was applied to detect the expression level of microRNA-144 after lncRNA-TUG1 was silenced. Changes of c-Met mRNA and protein expressions was detected by qRT-PCR and western-blot test. Results The expression level of lncRNA-TUG1 in GC tissue was significant higher than that in para-C tissue (P < 0.05) and the high expression level of lncRNA-TUG1 in GC tissue was significantly correlated with tumor lymph nodes metastasis and advance TNM phasing (P < 0.05). The transferring and invading capacity of SGC-7901 cells was highly inhibited after being transfected by lncRNA-TUG1 specific siRNA (P < 0.05). The results of qRT-PCR and western-blot proved that the expression of microRNA-144 was significantly boosted and the expression level of c-Met mRNA and protein was inhibited after lncRNA-TUG1 was silenced (P < 0.05). Conclusions lncRNA-TUG1 shows an up-regulated expression in GC tissue and that bears a correlation with clinicopathological features of malignant tumor. lncRNA-TUG1 can promote the transferring and invading capacity of GC by inhibiting the pathway of microRNA-144/c-Met.