Asian Pacific Journal of Tropical Medicine

Volume 10,Issue 7,2017 Table of Contents

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  • 1  Retraction notice to “Effect of phentolamine on myocardial extracellular matrix of cardiac remodeling in rats” [Asian Pac J Trop Med (2014) 645–649]
    Yi-Gang Yin Ru-Zhu Wang Zhong-Bao Ruan Li Zhu
    2017, 10(7).
    [Abstract](46) [HTML](0) [PDF 2.81 M](309)
    2  Research advances on the multiple uses of Moringa oleifera: A sustainable alternative for socially neglected population
    Raimunda S^amia Nogueira Brilhante Jamille Alencar Sales Vandbergue Santos Pereira Débora de Souza Collares Maia Castelo-Branco Rossana de Aguiar Cordeiro Célia Maria de Souza Sampaio Manoel de Araújo Neto Paiva Jo?o Bosco Feitosa dos Santos José Júlio Costa Sidrim Marcos F abio Gadelha Rocha
    2017, 10(7):621-630. DOI: 10.1016/j.apjtm.2017.07.002
    [Abstract](75) [HTML](0) [PDF 3.00 M](290)
    Moringa oleifera Lam (Moringaceae) is a plant with high nutritional and medicinal value. Native to India, it is now widely distributed throughout tropical and subtropical regions of the world. Its different parts are sources of proteins, vitamins and minerals and present different pharmacological and biotechnological potential. Moreover, M. oleifera seeds are widely used in water and effluent treatment, for their coagulation, flocculation and sedimentation properties, their ability of improving water quality, by reducing organic matter and microbial load, with special applicability in intensive animal production systems, such as aquaculture. In addition, due to its high nutritional value and several medicinal properties, this tree may act as a nutritional and medical alternative for socially neglected populations. In this context, this review gathers information on M. oleifera, emphasizing its chemical constituents, nutritional, pharmacological and antimicrobial properties, applications in the treatment of water effluents, and ecological and social aspects.
    3  Alternate paradigms on Zika virus-related complications: An analytical review
    Baburajan Radha Gnanaraj Muniraj
    2017, 10(7):631-634. DOI: 10.1016/j.apjtm.2017.07.003
    [Abstract](49) [HTML](0) [PDF 2.81 M](277)
    The proportion of the reported cases of Zika virus (ZIKV) infection reached the status of a pandemic. Numerous studies are being conducted on the isolation of ZIKV strains from various epidemics, diagnosis of the infections, various animal models and cell culture designs to study the pathogenesis of ZIKV in the attempts to find an effective ZIKV vaccine. This review focuses upon the ‘Off-Spectrum’ body of studies which analyses the epidemiology, pathogenesis and other attributes of ZIKV in the light of various dissident hypotheses.
    4  GC–MS analysis of volatile compounds of Perilla frutescens Britton var. Japonica accessions: Morphological and seasonal variability
    Bimal Kumar Ghimire Ji Hye Yoo Chang Yeon Yu Ill-Min Chung
    2017, 10(7):643-651. DOI: 10.1016/j.apjtm.2017.07.004
    [Abstract](46) [HTML](0) [PDF 2.82 M](328)
    Objective: To investigate the composition of volatile compounds in the different accessions of Perilla frutescens (P. frutescens) collected from various habitats of China and Japan. Methods: In the present study, the essential oil from the leaves of P. frutescens cultivars from China and Japan was extracted by hydro-distillation and the chemical composition and concentration of the volatile components present in the oils were determined by gas chromatography–mass spectrometry (GC–MS) analysis. Results: Among the volatile components, the major proportion was of perilla ketone, which was followed by elemicin and beta-caryophyllene in the Chinese Perilla cultivars. The main component in the oil extracted from the Japanese accessions was myristicin, which was followed by perilla ketone and beta-caryophyllene. We could distinguish seven chemotypes, namely the perilla ketone (PK) type, perilla ketone, myristicin (PM) type, perilla ketone, unknown (PU) type, perilla ketone, beta-caryophyllene, myristicine (PB) type, perilla ketone, myristicin, unknown (PMU) type, perilla ketone, elemicine, myristicin, beta-caryophyllene (PEMB) type, and the perilla ketone, limonene, betacryophyllene, myristicin (L) type. Most of the accessions possessed higher essential oil content before the flowering time than at the flowering stage. The average plant height, leaf length, leaf width of the Chinese accessions was higher than those of the Japanese accessions. Conclusion: The results revealed that the harvest time and geographical origin caused polymorphisms in the essential oil composition and morphological traits in the Perilla accessions originating from China and Japan. Therefore, these chemotypes with desirable characters might be useful for industrial exploitation and for determining the harvest time.
    5  Production, characterization and biological activities of acidic exopolysaccharide from marine Bacillus amyloliquefaciens 3MS 2017
    Samah A. El-Newary Abeer Y. Ibrahim Mohsen S. Asker Manal G. Mahmoud Mohamed E. El Awady
    2017, 10(7):652-662. DOI: 10.1016/j.apjtm.2017.07.005
    [Abstract](55) [HTML](0) [PDF 3.02 M](260)
    Objective: To evaluate in-vitro antioxidant, anti-inflammatory and antitumor abilities against human breast adenocarcinoma (MCF7) and human prostate cancer (PC3) as well as the suppressor effect of bacterial exopolysaccharide (BAEPS) on Ehrlich ascites carcinoma (EAC). Methods: In-vitro antioxidants characters of BAEPS were determined using various methods, while anti-inflammatory activity was estimated against cyclooxygenase (COX-1 and COX-2). In-vitro study, anticancer against MCF7 and PC3 were assessed by the mitochondrial dependent reduction of yellow MTT. In in-vivo study against EAC progression, mice were inoculated with EAC cells and then were orally administered BAEPS at 200 mg/kg after 24 h (equals to 0.10 of determinedLD50)/10 d. Results: BAEPS was acidic exopolysaccharide contained uronic acid (12.3%) and sulfate (22.8%) with constitution of glucose, galactose and glucuronic acid in a molar ratio 1.6:1.0:0.9, respectively, with a molecular mass of 3.76 ×104/sup>g/mol. BAEPS appeared potent antioxidant characters as free radical scavenging, oxygen reactive species scavenging and metal chelation, while its reducing power was low. BAEPS showed selective anti-inflammatory activity against COX-2 than COX-1, COX-2 selective. BAEPS exhibited potent and selective effect to breast cell cancer MCF7, the death percentage was 65.20% with IC50= 70 μg/mL and IC90 = 127.40 μg/mL. BAEPS decreased counted viable EAC cells and induced non-viable cells. BAEPS improved all assessed hematological parameters. These improvements were reflected in the increasing median survival time and significant increment (P < 0.05) in life span. Conclusions: BAEPS has anti-tumor activity with a good margin of safety. The antitumor activity of BAEPS may be due to its content from sulfated groups and uronic acids and they have antioxidant and anti-inflammatory properties.
    6  Chemical analysis and in vitro antimicrobial effects and mechanism of action of Trachyspermum copticum essential oil against Escherichia coli
    Wei Huang Hai-Yan Song Qian Zhang Guang-Fa Liu
    2017, 10(7):663-669. DOI: 10.1016/j.apjtm.2017.07.006
    [Abstract](38) [HTML](0) [PDF 2.81 M](247)
    7  Solvent–solvent fractionations of Combretum erythrophyllum (Burch.) leave extract: Studies of their antibacterial, antifungal, antioxidant and cytotoxicity potentials
    Fanyana M. Mtunzi Ikechukwu P. Ejidike Imelda Ledwaba Aroke Ahmed Vusumzi E. Pakade Michael J. Klink Sekomeng J. Modise
    2017, 10(7):670-679. DOI: 10.1016/j.apjtm.2017.07.007
    [Abstract](138) [HTML](0) [PDF 3.59 M](452)
    Objective: To evaluate the biological activities of Combretum erythrophyllum (C. erythrophyllum) leaf extracts against infectious diseases' pathogenesis and their cytotoxicity potentials. Methods: Powdered leaf material (300 g) of C. erythrophyllum was extracted (1:10 w/v) using acetone to obtain the crude extract. Liquid–liquid fractionation was performed on the crude acetone extract (30 g) using solvents of different polarity. The bioautographic method was used to detect the inhibition of bacterial and fungal growth by active compounds present in the crude and fractions. The extracts were then tested on bacterial strains: Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa; fungal strains: Candida albicans (C. albicans), Cryptococcus neoformans, and Aspergillus fumigatus, by microtitre dilution method for MIC determination. Results: The extracts MIC values ranged between 0.08 and 2.50 mg/mL against the tested pathogens. Water fraction had the highest activity against bacteria strains, while the fungal assay revealed crude acetone extract and ethyl acetate fraction to be active against C. albicans (1.25 mg/mL), dichloromethane extract against C. albicans and A. fumigatus (0.16 mg/mL). Extract fractions showed a good antioxidant activity via DPPH, ABTS and hydroxyl radical scavenging assays, in the order: ethyl acetate > water > acetone > dichloromethane > hexane. The toxicity level of crude extract and fractions evaluated in Vero monkey kidney cells ranged from 34 to 223 μg/ mL, while doxorubicin (IC50= 7.19 mg/mL) served as the positive control. Conclusions: It can be concluded that the extracts of C. erythrophyllum are safe for medicinal use in folk medicine for treating infectious and stress related diseases.
    8  Association between FGFR4 gene polymorphism and high-risk HPV infection cervical cancer
    Ya-Ping Li Lin Zhang Yu-Liang Zou Yan Yu
    2017, 10(7):680-684. DOI: 10.1016/j.apjtm.2017.07.008
    [Abstract](46) [HTML](0) [PDF 2.81 M](238)
    Objectives: To discuss the association between FGFR4 gene polymorphism rs351855 (Glu388Aly) and the susceptibility and chemotherapeutic effect of cervical cancer infected by high-risk type HPV. Methods: A total of 162 patients with high-risk HPV cervical cancer and 162 healthy women were collected and the genotypes of the FGFR4 rs351855 locus were detected. The genotype distributions in the two groups were compared. The cervical cancer patients were divided into four groups which namely good therapeutic effect group and bad therapeutic effect, recurrence or metastasis and no recurrence or metastasis group respectively, and the risks of different genotype on the curative effect and prognosis were analyzed by Logistic regression. The survival time of patients with different genotypes was compared. Results: There was no statistic difference in FGFR4 rs351855 genotype distribution between the patients group and control group (P>0.05), among which the risk of chemotherapy failure on GA + AA patients was 3.257 times as much as that of the GG patients, and the risk of recurrence or metastasis of GA + AA patients was 2.783 times as much as that of the GG patients. For AA patients, the risk of chemotherapy failure and the risk of relapse and metastasis are 3.833 and 3.406 times, respectively, as much as that of the GG patients. The overall survival of GA and AA patients was shorterthanthat of the GG patients, and significant difference was found (χ2= 7.098, P = 0.029). The difference in overall survival between GA + AA patients and GG patients was almost statistically significant (χ2= 3.634, P = 0.057). Conclusions: The FGFR4 rs351855 polymorphism is not associated with the susceptibility of high-risk HPV cervical cancer, but patients with gene A was at higher risk of unfavorable chemotherapy prognosis compared with patients with GG.
    9  Mechanism of antagonistic effects of Andrographis paniculata methanolic extract against Staphylococcus aureus
    Roslinah Mohamad Hussain Zayan Nabilah Rasyidah Abd. Razak Wan Mazlina Md Saad Maimunah Mustakim
    2017, 10(7):685-695. DOI: 10.1016/j.apjtm.2017.07.009
    [Abstract](59) [HTML](0) [PDF 2.61 M](295)
    Objective: To investigate the effects of Andrographis paniculata (Burm.f.) Wall. Ex Nees (A. paniculata) on expressions and activities of catalase, superoxide dismutase and alkylhydroperoxide reductase C in Staphylococcus aureus (S. aureus) with respect to its survival in vitro. Methods: Antioxidative property of methanolic leaves extract of A. paniculata (0.06 mg/ mL). Minimum inhibitory concentration (MIC) was determined by its ability to reduce hydrogen peroxide H2O2 () toxicity against S. aureus ATCC 25923 [(3.8 ×108) cfu/mL]. Effects of the extract on expressions of katA (encoding catalase), sodA and sodM [encoding superoxide dismutases (SODs)], and ahpC [encoding alkylhydroperoxide reductase C (AhpC)] in S. aureus were determined by RT-qPCR and corresponding enzyme activity assays were performed. Nitroblue tetrazolium reduction (NBT) assay was performed to determine effects of the extract on intracellular and extracellular levels of O2- in S. aureus. Results: Cells challenged with 7.5 mmol/L H2O2 showed 0% survival in 30 min whereas 25% survived after treatment with the extract and H2O2. Cells that were treated with the extract alone had 43% survival in the same exposure period. Expressions of sodA and sodM genes in extract-treated cells were lowered 0.8-fold and 0.7-fold, respectively with decrease in total SOD activity of 26.8 U compared to untreated cells, 32.4 U (P < 0.05). In contrast, extract-treated S. aureus cells showed 3.3-fold increase in katA expression with corresponding increase in catalase activity of 1.828 U compared to untreated cells which was 1.248 U, (P < 0.05). More profoundly, ahpC expression was increased 61-fold in extracttreated cells, (P < 0.05) with corresponding increase in AhpC activity of 0.018 U compared to untreated cells, 0.012 U, (P < 0.05). Extract-treated cells had significantly lower intra- and extracellular O2- levels with absorbance readings (A575 nm) of 0.340 and 0.524 compared to untreated cells which were 0.516 and 0.928 (P < 0.05), respectively. Conclusions: Taken together these results suggest that the low MIC of A. paniculata methanolic leaves extract (0.06 mg/mL) reduce H2O2 toxicity and more importantly, was in itself effectively inhibitory against S. aureus. Further, our observations suggest that a probable mode of its inhibitory mechanism against S. aureus is by reducing total SOD activity through downregulation of sodA and sodM expressions.
    10  Effect of Taoren Quyu Decoction on human endometrial cells and its anti-endometriosis activity in rats
    Hai-Zhi Liu Xiao-Xue Han Jia Liu Feng-Cheng Zhu Rui-Man Li
    2017, 10(7):696-700. DOI: 2017; 10(7): 696–700
    [Abstract](65) [HTML](0) [PDF 3.00 M](276)
    Objective: To study the effect of Taoren Quyu Decoction (TQD) on endometrial cells in patients with endometriosis (EMs) and EMs in rats. Methods: A total of 60 female Wistar rats were randomly divided into 4 groups, namely, normal group, model group, positive group and TQD group, each group having 15 rats. Except the normal group, EMs model was established in the other three groups by transplanting the rat autologous endometrium. After 4 weeks of intragastric administration, blood, eutopic and ectopic endometrial tissues of rats in each group were collected to detect the serum levels of estrogen (E2), cancer antigen 125 (CA125), endometrial antibody (EMAb), and expressions of microvessel density (MVD), vascular endothelial growth factor (VEGF) and angiopoietin (Ang-2). The volume of endometriosis cyst was determined simultaneously. For the in vitro culture of human endometrial cells, 4 groups, namely, normal group, model group, positive group and TQD group were used. The positive group and TQD group were treated with danazol and TQD respectively. Then 24 h after the treatment, the expressions of survivin and tumor suppressor gene (p53) of each group were detected. Results: The volumes of the endometriosis cysts in the positive group and the TQD group were significantly reduced compared with the model group (P < 0.05). The serum levels of E2, CA125 and EMAb, and the expressions of MVD, VEGF and Ang-2 in the model group were significantly increased compared with the normal group (P < 0.05); while they were all significantly reduced in the positive group and TQD group (P < 0.05). Compared with the normal group, the expression of survivin in the model group was significantly up-regulated (P < 0.05), and expression of p53 was significantly reduced (P < 0.05); compared with the model group, the expressions of survivin in the positive and TQD groups were significantly decreased (P < 0.05), and expression of p53 was significantly up-regulated (P < 0.05). The difference between positive group and TQD group was not statistically significant (P > 0.05). Conclusions: TQD has a significant anti-EMs effect, and its mechanism of action may be related to anti-angiogenesis and promoting apoptosis of ectopic endometrial cell.
    11  In vitro inhibitory analysis of consensus siRNAs against NS3 gene of hepatitis C virus 1a genotype
    Imran Shahid Waleed Hassan AlMalki Mohammed Wanees AlRabia Mohammed Hasan Mukhtar Shaia Saleh R. Almalki Saad Ahmed Alkahtani Sami S. Ashgar Hani S. Faidah Muhammad Hassan Hafeez
    2017, 10(7):701-709. DOI: 10.1016/j.apjtm.2017.07.011
    [Abstract](49) [HTML](0) [PDF 2.22 M](252)
    Objective: To explore inhibitory effects of genome-specific, chemically synthesized siRNAs (small interference RNA) against NS3 gene of hepatitis C virus (HCV) 1a genotype in stable Huh-7 (human hepatoma) cells as well as against viral replication in serum-inoculated Huh-7 cells. Methods: Stable Huh-7 cells persistently expressing NS3 gene were produced under antibiotic gentamycin (G418) selection. The cell clones resistant to 1 000 mg antibiotic concentration (G418) were picked as stable cell clones. The NS3 gene expression in stable cell clone was confirmed by RT-PCR and Western blotting. siRNA cell cytotoxicity was determined by MTT cell proliferation assay. Stable cell lines were transfected with sequence specific siRNAs and their inhibitory effects were determined by RT-PCR, realtime PCR and Western blotting. The viral replication inhibition by siRNAs in serum inoculated Huh-7 cells was determined by real-time PCR. Results: RT-PCR and Western blot analysis confirmed NS3 gene and protein expression in stable cell lines on day 10, 20 and 30 post transfection. MTT cell proliferation assay revealed that at most concentrated dose tested (50 nmol/L), siRNA had no cytotoxic effects on Huh-7 cells and cell proliferation remained unaffected. As demonstrated by the siRNA time-dependent inhibitory analysis, siRNA NS3-is44 showed maximum inhibition of NS3 gene in stable Huh-7 cell clones at 24 (80%, P = 0.013) and 48 h (75%, P = 0.002) post transfection. The impact of siRNAs on virus replication in serum inoculated Huh7 cells also demonstrated significant decrease in viral copy number, where siRNA NS3- is44 exhibited 70% (P < 0.05) viral RNA reduction as compared to NS3-is33, which showed a 64% (P < 0.05) decrease in viral copy number. siRNA synergism (NS3- is33 + NS3-is44) decreased viral load by 84% (P < 0.05) as compared to individual inhibition by each siRNA (i.e., 64%–70% (P < 0.05)) in serum-inoculated cells. Synthetic siRNAs mixture (NS5B-is88 + NS3-is33) targeting different region of HCV genome (NS5B and NS3) also decreased HCV viral load by 85% (P < 0.05) as compared to siRNA inhibitory effects alone (70% and 64% respectively, P < 0.05). Conclusions: siRNAs directed against NS3 gene significantly decreased mRNA and protein expression in stable cell clones. Viral replication was also vividly decreased inserum infected Huh-7 cells. Stable Huh-7 cells expressing NS3 gene is helpful to develop anti-hepatitis C drug screening assays. siRNA therapeutic potential along with other antiHCV agents can be considered against hepatitis C.
    12  Clinical study inpatient-reported outcomes after binocular implantation of aspheric intraocular lens of different negative spherical aberrations
    Xu-Hua Song Wen-Zhe Li Yan Dou Qiang Wu
    2017, 10(7):710-713. DOI: 10.1016/j.apjtm.2017.07.012
    [Abstract](37) [HTML](0) [PDF 3.01 M](307)
    Objective: To compare patient-reported outcomes after implantation of the ZA9003 intraocular lens (IOLs), or the MCX11 ASP IOLs or the spherical IOLs (HQ-201HEP). Methods: Prospective nonrandomized controlled trial was used. A total of 105 patients (210 eyes) were divided into three groups according to the type of IOLs: ZA9003 (35 patients, 70 eyes), MCX11 ASP (35 patients, 70 eyes) or HQ-201HEP (35 patients, 70 eyes). The main outcome was scores of Catquest nine-item short-form questionnaire. Additional outcome was best corrected visual acuities, spherical aberration (SA) and total higher-order aberrations (HOAs). Results: The global score was significantly lower in the spherical IOL group than the aspherical IOL group of −020 mm SA (P < 0.05) and the aspherical IOL group of −027 mm SA (P < 0.05), and no significant difference was found in the global score between the aspherical IOL group of −020 mm SA than the aspherical IOL group of −027 mm SA (P > 0.05). Significant differences were also found in question 2, question 5, question 6 and question 8 between the spherical IOLs and the aspherical IOLs. Conclusion: Implantation of an aspherical IOL could improve vision-related quality of life compared with a spherical IOL. However, there were no statistically significant differences in vision-related quality of life between aspheric IOLs with different negative spherical aberrations.
    13  Effect of axial vertical vibration on degeneration of lumbar intervertebral discs in modified bipedal rats: An in-vivo study
    Xiao Liang Hao Shen Wei-Dong Shi Shan Ren Wei Jiang Hao Liu Peng Yang Zhi-Yong Sun Jun Lin Hui-Lin Yang
    2017, 10(7):714-717. DOI: 10.1016/j.apjtm.2017.07.014
    [Abstract](30) [HTML](0) [PDF 2.81 M](243)
    Objective: To assess the effects of axial vibrations on gene expression and lumbar intervertebral disc degeneration in vivo. Methods: A modified bipedal rat model was established using a brachial plexus rhizotomy approach to imitate human upright posture. The experimental animals were randomly divided into three groups: control, vertical vibration, and whole-body vibration. Gene expression in degeneration of the intervertebral discs was assessed by reverse transcription-quantitative polymerase chain reaction. Results: The expression of aggrecan, Col1α1, Col2α1, and decorin were shown to be up-regulated in 14-week-old rats in the vertical vibration and whole-body vibration groups, whereas biglycan and versican expression was down-regulated in 14-week-old rats of the two experimental groups. Furthermore, biglycan and versican expression levels were shown to be lower in the whole-body vibration group than in the vertical vibration group (P < 0.05). Conclusions: This in-vivo study demonstrated that vibrations can influence the expression of anabolic genes. Furthermore, whole-body vibrations seem to have a greater effect in this regard than vertical vibrations. A new method is expected to relieve the low back pain of the patients through our research.
    14  First report on molecular characterization of Leishmania species from cutaneous leishmaniasis patients in southern Khyber Pakhtunkhwa province of Pakistan
    Mubbashir Hussain Shahzad Munir Sultan Ayaz Bahar Ullah Khattak Taj Ali Khan Niaz Muhammad Muhammad Anees Hazir Rahman Muhammad Qasim Muhammad Ameen Jamal Irfan Ahmed Kashif Rahim Humaira Mazhar Noha Watanay Mohamed Kasbari
    2017, 10(7):718-721. DOI: 10.1016/j.apjtm.2017.07.015
    [Abstract](71) [HTML](0) [PDF 3.01 M](294)
    Objective: To report presence of Leishmania major in Khyber Pakhtunkhwa of Pakistan, where cutaneous leishmaniasis (CL) is endemic and was thought to be caused by Leishmania tropica only. Methods: Biopsy samples from 432 CL suspected patients were collected from 3 southern districts of Khyber Pakhtunkhwa during years 2011–2016. Microscopy on Giemsa stained slides were done followed by amplification of the ribosomal internal transcribed spacer 1 gene. Results: Leishmania amastigotes were detected by microscopy in 308 of 432 samples (71.3%) while 374 out of 432 samples (86.6%) were positive by ribosomal internal transcribed spacer 1 PCR. Subsequent restriction fragment length polymorphism confirmed L. tropica in 351 and L. major in 6 biopsy samples. Conclusions: This study is the first molecular characterization of Leishmania species in southern Khyber Pakhtunkhwa. It confirmed the previous assumptions that anthroponotic CL is the major CL form present in Khyber Pakhtunkhwa province. Furthermore, this is the first report of L. major from a classical anthroponotic CL endemic focus identified in rural areas of Kohat district in southern Khyber Pakhtunkhwa.

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