Asian Pacific Journal of Tropical Medicine

673-681

Authors: Lin Zhang, Anjaneya S. Ravipati, Sundar R Koyyalamudi, Sang Chul Jeong, Narsimha Reddy, John Bartlett, Paul T. Smith, Mercedes de la Cruz, Maria Cândida Monteiro, Ángeles Melguizo, Ester Jiménez, Francisca Vicente

Objective To evaluate in vitro antimicrobial activities of selected 58 ethno-medicinal plant extracts with a view to assess their therapeutic potential. Methods A total of 58 traditional Chinese medicinal plants were carefully selected based on the literature review and their traditional use. The antimicrobial activities of ethanol extracts of these medicinal plants were tested against fungi (Aspergillus fumigatus), yeast (Candida albicans), gram-negative (Acinetobacter baumannii and Pseudomonas aeruginosa) and gram-positive bacteria (Staphylococcus aureus). The activities were tested at three different concentrations of 1.00, 0.10 and 0.01 mg/mL. The data was analysed using Gene data Screener program. Results The measured antimicrobial activities indicated that out of the 58 plant extracts, 15 extracts showed anti-fungal activity and 23 extracts exhibited anti-bacterial activity. Eight plant extracts have exhibited both anti-bacterial and anti-fungal activities. For instance, Eucommia ulmoides, Polygonum cuspidatum, Poria cocos and Uncaria rhyncophylla showed activity against both bacterial and fungal strains, indicating their broad spectrum of activity. Conclusions The results revealed that the ethanol extracts of 30 plants out of the selected 58 possess significant antimicrobial activities. It is interesting to note that the findings from the current study are consistent with the traditional use. A clear correlation has also been found between the antimicrobial activity and the flavonoid content of the plant extracts which is in agreement with the literature. Hence, the results presented here can be used to guide the selection of potential plant species for the isolation and structure elucidation of novel antimicrobial compounds in order to establish the structure-activity relationship. This in turn is expected to lead the way to the discovery of novel antimicrobial agents for therapeutic use.

689-692

Authors: KN Sahare, V Singh

Objective To evaluate the possible antifilarial effect of ethyl acetate extract of Vitex negundo (Verbenaceae) leaves against Setaria cervi filarial parasite in vitro. Methods In vitro screening was done by the method of motility inhibition and MTT reduction assay with concentrations of 0.03 to 1.00 mg/mL for 2 to 24 h incubation periods respectively, for possible antifilarial effect by comparing with control. Results In motility assay, complete inhibition of motility was observed and in MTT reduction assay which gave >50% reduction for concentrations 0.20, 0.50 and 1.00 mg/mL at 10, 6 and 2 h incubation periods respectively in a dose dependent manner (P<0.05). An antifilarial effect imparted by plant extract was found to be a function of their relative concentrations. Inhibitory concentration (IC50) for the plant extract was found to be 0.16 mg/mL. Conclusions The present study recorded significant antifilarial effect of Vitex negundo plant extract and contributed toward the development of database for novel drug candidates for lymphatic filariasis.

699-702

Authors: Sirima Kitvatanachai, Pochong Rhongbutsri

Objective To provide baseline information of parasitic infections in 3 suburban government schools, Lakhok subdistrict of Muang Pathum Thani, Thailand. Methods This study was conducted between May-June 2010 using simple direct smear and modified formalin ether and from a population of 1 253 in 3 suburban government schools. Results Total samples of 202 registered and participated. The average of prevalence of infection from 3 schools was 13.9%, there were 13.7%, 14.3% and 13.9% in N, S and R school, respectively with no significant difference between schools (P>0.05). The infection rates did not show significant difference between genders (P>0.05). The highest rate of infection was 20.4% in Pathom 2 (8 years) students and the lowest was 4% in Pathom 1 (7 years) with statistically difference between age groups (P<0.05). The highest prevalence of pathogenic protozoa was Giardia lamblia (G. lamblia) which was found in 50% of infected cases, followed by 25% of Entamoeba histolytica (E. histolytica) and Blastocystis hominis (B. hominis). The highest prevalence of non-pathogenic protozoa was Endolimax nana (E. nana) which was found in 88.9%, followed by 11.1% of Entamoeba coli (E. coli). Mixed infections between Blastocystis hominis (B. hominis) and Endolimax nana (E. nana) were reported at 7.1%. The only helminthic infection found in this study area was hookworm, found in 1 student (3.8%). The formalin ether concentration technique showed a higher efficacy of detection (78%-100%) than the simple direct smear method (0%-50%). Conclusions Surveillance of Protozoan infections may need to be focused on suburban areas.

709-712

Authors: Ya-Jun Lu, Gang Lu, Da-Zhong Shi, Li-Hua Li, Sai-Feng Zhong

Objective To predict structure and function of translationally controlled tumor protein (TCTP) from Spirometra mansoni by bioinformatics technology, and to provide a theoretical basis for further study. Methods Open reading frame (ORF) of EST sequence from Spirometra mansoni was obtained by ORF finder and was translated into amino acid residue by DNAclub. The structure domain was analyzed by Blast. By the method of online analysis tools: Protparam, InterProScan, protscale, SignalP-3.0, PSORT II, BepiPred, TMHMM, VectorNTI Suite 9 packages and Phyre2, the structure and function of the protein were predicted and analyzed. Results The results showed that the EST sequence was Sm TCTP with 173 amino acid residues, theoretical molecular weight was 19 872.0 Da. The protein has the closest evolutionary status with Clonorchis sinensis, Schistosoma mansoni, and Schistosoma japonicum. Then it had no signal peptide site and transmembrane domain. Secondary structure of TCTP contained two α -helices and eight β -strands. Conclusions Sm TCTP was a variety of biological functions of protein that may be used as a vaccine candidate molecule and drug target.

718-722

Authors: Al-Bahry SN, Al-Mashani BM, Al-Ansari AS, Elshafie AE, Mahmoud IY

Objective To screen for Escherichia coli (E. coli) resistant to tetracycline, followed by identification of tet efflux genes by polymerase chain reaction (PCR). In addition, detection of tetracycline residues in chicken livers and kidneys were conducted using high performance liquid chromatography-tandem quadrupole mass spectrometry (HPLC-MS-MS). Methods Strains of E. coli were isolated from samples of chicken colon and screened for tetracycline resistance. Tetracycline genes conferring resistance (Tcr) were detected by polymerase chain reaction (PCR). Most of the isolates were resistant to tetracycline (97.9%). Results PCR analysis indicated that Tcr E. coli R-plasmids contained tet(A), tet(B) and a combination of both efflux genes. None of the isolates contained other efflux tet genes tet (C, D, E and Y). High performance liquid chromatography-tandem quadrupole mass spectrometry (HPLC-MS-MS), a sensitive technique, was used to detect residues of chlortetracycline (CTC), oxytetracycline (OTC), doxycycline (DC) in chicken livers and kidneys. The samples containing tetracycline residues were at 0.13-0.65 pg/μL levels. Conclusions Tetracycline and other antibiotics are commonly used in the poultry and meat production industry for prevention of microbial infections. Multiple antibiotic resistant bacteria in Oman have increased to alarming levels, threatening public health, domestic and may have adverse effect on environment.

728-731

Authors: Gomathi Sivaramakrishnan, Balaji Subramanyam, Ponnuraja C, Vanaja Kumar

Objective To evaluate luciferase reporter phage (LRP) phAE85 in rapid detection of rifampicin resistance in a region where TB is endemic. Methods One hundred and ninety primary isolates on Lowenstein-Jensen medium were tested. Middlebrook 7H9 complete medium with and without rifampicin at 2 μg/mL was inoculated with standard inoculum from suspensions of the clinical isolate. After incubation for 72 h, LRP was added. Following 4 h of further incubation, light output from both control and test was measured as relative light units. Strains exhibiting a reduction of less than 50% relative light units in the drug containing vial compared to control were classified as resistant. Results were compared with the conventional minimum inhibitory concentration method (MIC) of drug susceptibility testing. Results The two methods showed high level of agreement of 97% (CI 0.94, 0.99) and P value was 0.000 1. The sensitivity and specificity of LRP assay for detection of rifampicin resistance were 91% (CI 0.75, 0.98) and 99% (CI 0.95, 1.00) respectively. Time to detection of resistance by LRP assay was 3 d in comparison with 28 d by the minimum inhibitory concentration method. Conclusions LRP assay with phAE85 is 99% specific, 91% sensitive and is highly reproducible. Thus the assay offers a simple procedure for drug sensitivity testing, within the scope of semi-automation.

739-742

Authors: Tao Wang, Tian-An Liao, Shao-Bo Zhong

Objective To study the role of bone marrow mesenchymal stem cells (BMSCs) in construction of vascularized engineered tissue. Methods hVEGF165 was amplified via RT-PCR before recombinant with pShuttle- green fluorescence protein;green fluorescent protein (GFP)-CMV. Then the recombinant shuttle plasmid was transfected into BMSCs with LipofectamineTM 2000 for packaging and amplifying. hVEGF165 mRNA expression in BMSCs cells was tested. Results: The sequence of hVEGF165 in pShuttle-GFP-hVEGF165 plasmid was confirmed by double-enzyme cleavage method and sequencing. hVEGF165 was highly expressed in BMSCs. Conclusions The GFP/hVEGF165 recombinant plasmid vector was constructed successfully and expressed effectively in host cells, which may be helpful for discussing the possibility of the application of VEGF165-BMSCs in tissue engineering and ischemic disease cure.

748-752

Authors: Yuan-Huan Xiong, Zhen Yuan, Li He

Objective To investigate the effects of estrogen (E2) level on regulatory T cells (Treg) in peripheral blood during pregnancy. Methods A total of 30 healthy non-pregnant women were selected as control group, 90 pregnant women of early, middle and late pregnancy and 30 postpartum women at 1 month after parturition were selected as experimental groups including early pregnancy group, middle pregnancy group and late pregnancy group; the proportions of CD4+CD25+ Treg and CD4+CD25+CD127− Treg among CD4+T cells were detected by flow cytometry; the serum estrogen content in peripheral blood was detected by electrochemical immune luminescence method. Results E2 level was coincident with the change of Tregs number during pregnancy. The estrogen content in peripheral blood increased gradually from early pregnancy to late pregnancy, then decreased significantly after parturition, and the level at 1 month after parturition down to the level in non- pregnancy group (P>0.05); the level of E2 in pregnancy groups were significantly higher than those in non- pregnancy group (P<0.01); and there were significant differences among early pregnancy group, middle pregnancy group and late pregnancy group (P<0.05). The proportions of CD4+ CD25+ Treg and CD4+ CD25+ CD127− Treg in pregnancy groups were significantly higher than those in non- pregnancy group (P<0.05), but decreased significantly after parturition, and there was no significant difference between non- pregnancy group and postpartum women group (P>0.05); the proportions in middle and late pregnancy groups were significantly higher than those in early pregnancy group (P<0.05), but decreased slightly in late pregnancy group, there was no significant difference between late pregnancy group and middle pregnancy group (P>0.05). There was correlation between Tregs number with estrogen level during pregnancy. The proportion of CD4+ CD25+ Treg and CD4+ CD25+ CD127− Treg were positively correlated with estrogen level. Conclusions High proportion of CD4+ CD25+ Treg and CD4+CD25+CD127− Treg is closely related to the high level of E2 during pregnancy. It suggested that high level of estrogen may induce an increase of CD4+ CD25+ Treg in peripheral blood, and then influence the immune function of pregnant women. The results of this experiment might play an important role of estrogen in immune-modulation during pregnancy.

757-761

Authors: Jian-Hao Yang, Zheng-Chen Li, Wei-Dong Kong, Wu Zhang, Ying-Ping Jia, Yue-Lan Zhang, Lin-Bo Liu, Xue-Ping Han

Objective To investigate effect of orthodontic force on inflammatory periodontal tissue remodeling and expression of IL-6 and IL-8 in rats. Methods Eighty SD rats were randomly divided into 4 groups, blank control group (group A) with 5 rats, treatment normal group (group B) with 25 rats, inflammation control group (group (group C) with 25 rats, inflammation treatment group (group D) with 25 rats. Immunohistochemistry and histomorphometric analysis was performed to measure the expression of IL-6, IL-8 and the first molar to the recent movement in the distance. Results The expression of IL-8 reached a maximum on day 5 and declined thereafter in group B; the expression of IL-6 reached a maximum on day 5 in group B. The expression of IL-6 and IL-8 was gradually weakened with time in group C. The expression of IL-6 and IL-8 were high, and reached a maximum on day 5 and declined thereafter in group D. AD of positive cells in group D were higher than group B at each time point (P<0.05). The time which 0.49 N orthodontic force was loaded was longer, orthodontic tooth movement distance was greater. Movement distance in group D were longer than group B (P<0.05). Conclusions Orthodontic force as well as inflammatory stimulus can evoke the expression of IL-6 and IL-8. Under the combined effects of inflammation and orthodontic force, the expression of IL-6, IL-8 will increase.