Asian Pacific Journal of Tropical Medicine

925-933

Authors: Ayyakkannu Purushothaman, Elumalai Nandhakumar, Panchanatham Sachdanandam

Objective To investigate the bioactive constituents of Shemamruthaa (SM), a herbal combination and its therapeutic effects on the mitochondrial functions with reference to lipid peroxidation (LPO), antioxidant status, citric acid cycle enzymes and electron transport chain enzymes in mammary tissues of 7,12-dimethylbenz(a)-anthracene (DMBA) induced mammary carcinoma in rat model. Methods Adult female Sprague-Dawley rats were used for the study and were divided into four groups. Group I served as control and Group II rats were induced mammary carcinoma by administration of DMBA (25 mg/kg b.w.) orally. The normal and cancer-induced rats (Group III) were treated with SM (400 mg/kg b.w./day) orally by gastric incubation for 14 days. Group IV rats served as SM-treated control animals. Results Cancer-induced rats showed a considerably increased level of LPO with concomitant decreased levels of antioxidants, citric acid cycle enzymes, electron transport chain enzymes and cytochrome contents in the mammary tissue. Treatment with SM brought back the aforementioned biochemical parameters to near normal. Conclusions From the results, it can be inferred that Shemamruthaa possesses significant anticancer effect through its role in attenuation of LPO, prevention of membrane damage and restoring membrane integrity.

942-946

Authors: A. Nematollahi, G.H. Moghaddam, R. Jaafari, J. Ashrafi Helan, M. Norouzi

Objective To determine Neospora caninum (N. caninum) as a cause of bovine abortion in dairy cows by ELISA, PCR and Pathological methods in Tabriz, Northwest of Iran. Methods For study of outbreak of neosporosis, blood samples were collected from 76 Holstein aborted dairy cows in Tabriz (Northwest Iran). Antibodies to N. caninum were assayed by using a commercially ELISA kit (IDEXX, USA). IgG against N. caninum were found in 14 (18.4%) cases. Aborted fetuses of these seropositive dams were proposed for histopathological and molecular investigations. Brains, spinal cords and placentas of the fetuses were fixed in 10% buffered formalin for histopathology. Also 5-10 g of brain tissue was sampled for DNA extraction. In 6 out of 14 (42.8%) fetuses, brain tissue was positive in PCR. All dams of these fetuses were serologically seropositive in ELISA test. Results Histopathologically, the lesions consistent with N. caninum were observed in brains, spinal cords and placenta of all fetuses. Lesions in CNS included severe congestion, perivascular and perineuronal edema, status spongiosis, perivascular cuffing, focal gliosis, nourophagy and focal necrosis. There were some Neospora-like cysts in brain. In placentas, severe congestion, perivascular infiltration of mononuclear cells, vascular thrombosis, focal placentitis and necrotic foci in cotyledons were noticed. Conclusions The results of present study agreed the results of similar studies about serological, histopathological and molecular results of other studies about neosporosis and it seems to support the outbreak of N. caninum-associated abortion in dairy cows in Tabriz (Northwest Iran).

952-959

Authors: Olajire A. Adegoke, Olalekan A. Orokotan

Objective To evaluate the success rate of tuberculosis intervention programme at a specialist hospital in Ibadan, Nigeria through a retrospective study as well as carry out physicochemical evaluation of anti-tuberculous agents as a way of eliminating drug-related failure. Methods The retrospective study involved the use of quarterly tuberculosis central register at the Government Chest Hospital, Ibadan between 1st quarter (2003) to 4th quarter (2009). Relevant data were extracted from these register with the aid of data collection forms. The basic physicochemical analyses of the drugs given to the patients were also carried out using the International Pharmacopoeia methods. Results All the drugs examined for their physicochemical properties passed the International Pharmacopeia recommended tests. A total number of 1 260 patients enrolled at the hospital were assessed through case notes. This comprises of 59.4% males of which 69.23% new cases were also males. There was a significant (P<0.05) patient enrollment across the quarters for the seven years. An overall 80.24% cure rate over the 7-period was obtained which is less than the WHO target of 85%. Cure rates were better in females than males. Failure treatment outcomes such as positive (1.51%), deaths (8.73%), defaulted (3.33%) and transferred out (5.95%) were recorded though not statistically significant (P>0.05). Failure rates in all categories were higher in males than females (P>0.05). Conclusions More enlightenment and counseling is still required to meet up with the target for TB control.

964-967

Authors: Gao-Feng Xu, Wan-Fu Xie

Objective To investigate the influence and possible mechanism of ERBB2 expression on the invasiveness of glioma cells. Methods Glioma TJ905 cells were separated and cultured. ERBB2 shRNA and overexpressing vectors were constructed, which were then transfected. The ERBB2 expression was up-regulated or down-regulated. Changes of invasiveness of TJ905 cells were detected by Transwell assay, and the expressions of matrix metalloprotease (MMP)-2 and MMP-9 were measured by Western blot. Results ERBB2 shRNA transfection vector could effectively inhibit expression of ERBB2; while ERBB2 overexpressing vector transfection could significantly improve the expression of ERBB2 in TJ905 cells. Transwell assay showed that when ERBB2 expression was down-regulated, the invasiveness of TJ905 cells was notably decreased; when ERBB2 expression was up-regulated, the invasiveness of TJ905 cells was markedly increased. Meanwhile, Western blot indicated that down-regulating ERBB2 inhibited the expression of MMP-2 and MMP-9, while up-regulating ERBB2 enhanced their expressions. Conclusions ERBB2 expression is closely related to the invasiveness of glioma TJ905 cells.

972-976

Authors: Yong-Gang Tan, Yong-Feng Zhang, Chang-Jun Guo, Min Yang, Man-Yin Chen

Objective To investigate the differential expression of microRNA (miRNA) in colon between ulcerative colitis (UC) and ulcerative colitis related colorectal cancer (UCRCC). Methods An UC mouse model was built by dextran sodium sulfate, and an UCRCC mouse model by dextran sodium sulfate and 1,2-diformylhydrazine. RNAs were extracted from the colon, purified and hybridized with fluorescence-labeled miRNA oligonucleotide gene chip. Real-time fluorescence quantitative PCR was used to verify the expression variation of miRNA. SAM was employed for the data analysis. Results The up-regulated miRNAs in colon cancer included has-miR-194, has-miR-215, has-miR-93, has-miR-192, has-miR-92a, has-miR-29b, and has-miR-20a (median false discovery rate<5%), while the down-regulated miRNAs were has-miR-1231, has-miR-195, has-miR-143, and has-miR-145 (median false discovery rate<5%). Conclusions Significant differential expression of miRNA was found between the UC mouse and UCRCC mouse, which may be related to the onset, erosion and transfer of colorectal cancer.

982-985

Authors: Na Li, Hua-Qiong Huang, Gen-Sheng Zhang, Wei Cui

Objective To explore effect of 5-AZn-2 ′-deoxycytidine on proliferation of human lung adenocarcinoma cell line A549 in vitro. Methods Superoxide dismutase (SOD) activity was measured by hydroxylamine colorimetric method. Inhibition effect of 5-AZn-2′ deoxycytidylic acid at different concentration and different time on growth of A549 cell was determined by MTT assay. Methylene dioxyamphetamine (MDA) was measured by thiobarbituric acid colorimetric method. Effect of 5-AZn-2′ deoxycytidylic acid on apoptosis of A549 cell was determined by Hoechst 33258 dyeing detection. Results 5-AZn-2′ deoxycytidylic acid had significant inhibition effect on proliferation of A549 cells in vitro, and the inhibition was notably dependent on time and dosage during 48-72 h; SOD level was significantly lower than those of control group (P<0.05, P<0.01), MDA level was significantly higher than those in the control group (P<0.05, P<0.01). A549 cells began to be in apoptosis after using 5-AZn-2′deoxycytidylic acid. Conclusions 5- AZn-2′ deoxycytidylic acid has significant inhibition effect on growth of A549 cell, and can lead the change of lipid peroxidation. It indicates that the mechanism has relationship with A549 cell cycle tissue and induction factor of apoptosis.

990-994

Authors: Fu Hua, Chang-Hua Li, Hong Wang, Hong-Ge Xu

Objective To investigate the roles of COX-2, TNF-α, IL-6 in the pathogenesis of autoimmune-type recurrent spontaneous abortion (RSA). Methods RT-PCR was used to detect the mRNA of COX-2, TNF-α, IL-6 in the trophoblast cells of murine RSA and normal pregnant models. The COX-2, TNF-α, IL-6 protein expressions were determined by using immunohistochemisry staining method. The COX-2, TNF-α, IL-6 protein expressions were determined by ELISA. Results The embryo loss rates in experiment group was significantly higher than that in normal pregnancy control group, the expression of COX-2, TNF-α, IL-6 in the trophoblast cells of murine RSA and normal pregnant models. The expression of COX-2 in autoimmune-type recurrent spontaneous abortion was significantly lesser than in normal pregnant models. The expression of TNF-α, IL-6 in autoimmune-type recurrent spontaneous abortion was significantly higher than in normal pregnant models. There was a positively correlation between TNF-α and IL-6. There was no relationship between COX-2, TNF-α and IL-6. Conclusions The abnormal expression of COX-2, TNF-α and IL-6 may result in RSA.

999-1003

Authors: Ye Erbo lati. Ali Mira, Muhuyati, Wu-Hong Lu, Peng-Yi He, Zhi-Qiang Liu, Yu-Chun Yang

Objective To observe the expression changes of inflammatory markers TGF-β1, Smad3 and IL-6 in patients with atrial fibrillation (AF), and to explore the significance of TGF-β1 signaling pathway in the structural remodeling of AF. Methods The expression of TGF-β1, Smad3 and IL-6 in 50 cases with AF and 30 normal cases were detected by RT-PCR and ELISA. Results The TGF-β1, Smad3 and IL-6 mRNA and protein expression levels in patients with AF were significantly higher than that in the control group (P<0.05), but there was no significantly different between the paroxysmal AF group and the persistent AF group (P>0.05). The TGF-β1mRNA expression in the ⩾ 50 years subgroup was significantly higher than that in the <50 years subgroups, and it was higher in the NYHA III subgroup than in the I/II grade subgroup. It was also higher in the left ventricular ejection fraction (LVEF) <50% subgroup than in LVEF ⩾ 50% group, and it was significantly higher in the AF time ⩾ 36 months subgroup than that in <36 months subgroup (P<0.05). The Smad3 and IL-6 expressions in the in the LVEF <50% subgroup were both high that than that in LVEF ⩾ 50% group, and higher in the AF time ⩾ 36 months subgroup than that in <36 months subgroup (P<0.05). There were a positive correlation between TGF-β1, Smad3 and IL-6 (r=0.687, r=0.547). There were also a positive correlation between Smad3 and IL-6 mRNA (r=0.823). Conclusions AF is associated with inflammation, and the inflammation is also involved in the fibrillation and sustain of AF. The TGF-β1 signal pathway may be involved in the process of atrial structural remodeling in patients with AF, and iss related with the occurrence and maintenance of AF.

841-846

Authors: RC Adiele, BB Fakae, IU Isuzu

Objective To elucidate the pharmacological bases of oral administration of Securidaca longepedunculata (S. longepedunculata) root extract as an anthelmintic in folkloric medicine. Methods Albino mice were infected with infective third (L3) larval stage of Heligmosomoides polygyrus (H. polygyrus) by esophageal intubation. Following establishment of the adult worms in the intestine, the mice were treated with 0–2 000 mg/kg body weight (bw) of methanolic root extract of S. longepedunculata and 100 mg/kg bw of pyrantel embonate, the reference drug in vivo. Bioactivity and larvicidal effects of the extract were tested by exposing brine shrimps (Artemia salina) to 0.00–1.00 mg/mL and the L3 stage of Heligmosomoides contortus (H. contortus) and H. polygyrus to 0.00–2.50 mg/mL of the extract in vitro. Results The percentage yield of the extract was 7.13% w/w dry matter. The brine shrimps toxicity bioassay resulted in an LC50 of 74.18 μg/mL. The extract had a significant, dose–dependent larvicidal effect on the L3 stage of H. contortus and H. polygyrus with the terminal effect of 75% and 70% at the highest exposure concentrations, respectively. The extract however, did not affect the number of worm eggs per gram (epg) of fecal materials (P<0.05) and total worm burden (twb) of adult H. polygyrus in infected mice. Treatment with pyrantel embonate significant reduced both the fecal egg count and twb to 0 compared to the untreated control (P<0.05). Conclusions These results indicate that S. longepedunculata root extract contains potent bioactive compounds and has larvicidal effect on L3 stage of H. contortus and H. polygyrus, substantiating its use as anthelmintic in alternative medicine.

854-858

Authors: Selvaraj Premalatha, Kuppusamy Elumalai, Alagarmalai Jeyasankar

Objective To determine the larvicidal and pupicidal activites of Solanum trilobatum (S. trilobatum) leaf extracts against Aedes aegypti (Ae. aegypti), Culex quinquefasciatus (Cx. quinquefasciatus) and Anopheles stephensi (An. stephensi). Methods The larvicidal and pupucidal was determined at five different concentrations of 50, 100, 150, 200 and 250 ppm. Percentage of larval mortality was assessed after 48 h. Results Methanol extracts of S. trilobatum was found to be more susceptible against the larvae of Ae. aegypti, Cx. quinquefasciatus and An. stephensi at 250 ppm with a LC50 value of 125.43, 127.77 and 116.64 ppm respectively. Leaf methanol extracts of S. trilobatum also exhibited pupicidal and adult emergence properties. Conclusions These results suggested that the leaf extracts of S. trilobatum showed potential to be used as an ideal ecofriendly approach for the control of the Ae. Aegypti, Cx. quinquefasciatus and An. stephensi.

865-868

Authors: CH Houben, H Fleischmann, M Gückel

Objective To assess the prevalence of malaria parasitemia in north–east Nigeria and to evaluate the measures for the prevention of malaria. Methods A village in north–eastern Nigeria was selected for the cross sectional study at the height of the rainy season in October 2011. A total of 550 inhabitants of a hamlet were recruited for this study. After obtaining the consent individuals received a structured interview and were tested for malaria parasites in their blood films. Recruits testing positive for malaria were given a course of artemesinin–based combination therapy (ACT). Results A total of 497 inhabitants representing approximately 90 percent of the population participated: a quarter of the study group carried malaria parasitesexclusively Plasmodium falciparum (P. falciparum)–representing a P. falciparum parasite rate (PfPR) of 24.5%. Besides, 53/138 in the age group of 2 to < 10 years old children tested positive for P. falciparum representing a PfPR2–10 value of 38.4%. Malaria control measures were used in just under a third (157/497) of this cohort. Despite these measures 28/157 (17.8%) still tested positive for P. falciparum. Conclusions The malaria burden is overestimated for this region in north–east Nigeria. The findings support an intermediate pattern of malaria endemicity. The 30% bed nets coverage for malaria control is well below the WHO estimates for 2011.

876-883

Authors: MBS Donio, SFA Ronica, V Thanga Viji, S Velmurugan, J Adlin Jenifer, M Michaelbabu, T Citarasu

Objective To characterize the pharmacological importance of biosurfactants isolated from halophilic Bacillus sp BS3. Methods Halophilic Bacillus sp. BS3 was isolated from solar salt works, identified by 16S rRNA sequencing and was used for screening their biosurfactant production. Characters of the biosurfactant and their anticancer activity were analyzed and performed in mammary epithelial carcinoma cell at different concentrations. Results The biosurfactant were characterized by TLC, FTIR and GC–MS analysis and identified as lipopeptide type. GC–MS analysis revealed that, the biosurfactant had various compounds including 13–Docosenamide, (Z); Mannosamine, 9– and N,N,N',N'–tetramethyl. Surprisingly the antiviral activity was found against shrimp white spot syndrome virus (WSSV) by suppressing the viral replication and significantly raised shrimp survival (P<0.01). Anticancer activity performed in the mammary epithelial carcinoma cell at different concentrations of biosurfactants, among the various concentrations of biosurfactants such as 0.000 25, 0.002 5, 0.025, 0.25 and 2.5μg, the 0.25 μg concentration suppressed the cells significantly (P<0.05) to 24.8%. Conclusions Based on the findings, the present study concluded that, there is a possibility to develop eco–friendly antimicrobial and anticancer drugs from the extremophilic origin.

889-892

Authors: Shu–Jie Zhao, Dong Zhang, Shi–Ji Wang, Ying Chen, Jin–Feng Han, Yu–Shan Wang

Objective To analyze the effect of intestinal function–recovering decoction on multiple organ dysfunction syndrome in rats, and to investigate a novel solution to multiple organ dysfunction syndrome. Methods Multiple organ dysfunction syndrome was induced in 60 Sprague–Dawley rats by intestinal ischemia–reperfusion combined with cecal ligation and puncture. Then these rats were intragastrically administered physiological saline (group I, n=20), ampicillin (group II, n=20) or intestinal function–recovering decoction (group III, n=20). After treatment, serum malondialdehyde and superoxide dismutase levels were compared among three groups. Simultaneously, bacterial culture of various organ tissues was performed and bacterial and endotoxin translocation were observed. Results Compared with group I, serum malondialdehyde, alanine aminotransferase and aspartate aminotransferase levels were significantly decreased (all P<0.05) and serum superoxide dismutase level was significantly increased (P<0.05) in the group III. However, there were no significant differences in these indices between groups II and III (P>0.05). The rate of bacterial translocation in the groups II and III was significantly lower than in the group I (P<0.05), and no significant difference was observed between groups II and III (P>0.05). Conclusions Intestinal function–recovering decoction can significantly reduce endotoxin and bacterial translocation and stabilize enteral oxidative–antioxidative balance.

897-901

Authors: Li Wang, Zhi–Qiang Liu, Ying–Qian Huo, Li–Juan Yao, Xue–Gong Wei, Yun–Fang Wang

Objective To investigate the change of the hs–CRP, sVCAM–1, NT–proBNP levels of the patients with pregnancy–induced hypertension (PIH) syndrome. Methods A total of 200 patients with PIH were divided into mild, moderate and severe group, and 50 healthy pregnancy patients served as the control group. The serum sVCAM–1 levels were detected by enzyme–linked immunosorbent assay, hs–CRP were detected by immunity transmission turbidity, and NT–proBNP levels were determined by the colloidal gold method. Patients were treated with magnesium sulfate and nifudipine and the contrastive analysis was performed before and after treatment. And the pathological changes in placental of PIH patients were detected by hematoxylin–eosin staining at the same time. Results The hs–CRP, sVCAM–1, NT–proBNP levels of patients in the mild, moderate and severe PIH group were significantly higher than that in the control group (P<0.05). The hs–CRP, sVCAM–1, NT–proBNP levels in the severe group were significantly higher than the mild group and the moderate group, the difference was statistically significant (P<0.05). The hs–CRP, sVCAM–1, NT–proBNP of the moderate group were significantly higher than the mild group (P<0.05). There was a positive correlation between hs–CRP, sVCAM–1, NT–proBNP expression levels and the degree of the PIH. The expression of hs–CRP, sVCAM–1, NT–proBNP levels of the moderate and the severe group were significantly decreased (P<0.05). The number of placental villi and interstitial blood vessel in the moderate and severe PIH group were significantly less than the control group (P<0.05). Conclusions The increased levels of serum hs–CRP, sVCAM–1, NT–proBNP may be involved in the process of vascular endothelial cell injury of the PIH, and the hs–CRP, sVCAM–1, NT–proBNP can be used as the auxiliary index for diagnosis of PIH and determination of PIH severity.

908-911

Authors: Jia–Jia Wu, Shun–Chang Jiao

Objective To investigate the expression of hENTl and ERCC1 genes in tumor tissues non–small cell lung cancer (NSCLC). Methods Fresh non–small lung cancer specimens were transplanted into nude mice. Twenty mice were randomized into two groups: experimental group receiving gemcitabine plus cisplatin and control group receiving 0.9% physiological saline. The expressions of hENTl and ERCC1 mRNA in tumor tissue were detected by real–time fluorescent quantitative PCR. The volume of tumor, the weight of nude mice and tumor volume were respectively measured and calculated 2–3 times per week. Tissue samples were collected from NSCLC mice treated with gemcitabine plus carboplatin. Results The histological examination showed that many tumor cells were well preserved in nude mice. The rate of transplanted tumor cells was 86.7%. The concomitant treatment study showed that the rate of TV, RTV, T/C in GEM + DDP group was the lowest. LBP + DOC, DDP + DOC obviously influenced the body weight. Compared with NS group, DDP group, GEM group, the survival period and the level of hENTl of DDP+GEM group increased obviously, the level of ERCC1 decreased significantly (P<0.05). Conclusions The expression of hENT1 and ERCC1 genes in tumor tissues were closely correlated with the response to chemotherapy and prognosis of patients with NSCLC treated with gemcitabine plus cisplatin.

916-920

Authors: Ling–Zhi Liu, En–Ping Wu, Heng–Liang Liu

Objective To explore the relation between K469E gene polymorphism of intercellular adhesion molecular–1 (ICAM–1) and the recurrence of ACS and cardiovascular mortality. Methods A total of 185 patients with ACS hospitalized in Department of Cardiology in our hospital from Sep 2007 to Sep 2008 were selected as objectives. Polymerase chain reaction was used to analyze K469E gene polymorphism of ICAM–1. According to the genotypes, they were divided into two groups: group with K allele (KK+KE) and group without K allele (EE). The two groups were followed up prospectively for five years and blood lipid, blood pressure, blood glucose, recurrence and death of ACS were collected when the patients left hospital. The relation between ICAM–1 gene polymorphism and the recurrence of ACS and cardiovascular mortality was analyzed by Logistic regression. Results After long–term follow–up, it was found that ACS recurred on 71 cases (38.4%) and 10 cases died, among which 3 cases died of cardiovascular disease. The recurrence of ACS and cardiovascular mortality in group with K allele were remarkably higher than that in group without K allele (P<0.01). After multivariate Logistic regression adjusted ages, gender, weight indexes, TC, LDL–C, TG, smoking, drinking, family history of cardiovascular disease, history of hypertension and the severity of coronary artery disease, the risks of ACS recurrence and cardiovascular mortality in group with genotype KK+KE was 3.31 and 3.53 times of those in group with genotype EE respectively (P<0.01). When the independent variable of hypertension was introduced in regression analysis, the risks of ACS recurrence and cardiovascular mortality in group with K allele both decreased (P<0.05). When the independent variable of HDL–C was introduced, different genotypes of ICAM–1 weren't relevant with ACS recurrence and cardiovascular mortality (P>0.05). Conclusions K469E gene polymorphism of ICAM–1 was related to ACS recurrence and cardiovascular mortality, K allele probably an independent risky factor and hypertension and to which the level of HDL–C were closely related.

762-767

Authors: Zhen Yu, Li-Fen Chen, Ling Tang, Chang-Lin Hu

Objective To investigate the effects of adenovirus (Ad)-mediated hypoxia-inducible factor-1alpha (HIF-1α) gene on proliferation and differentiation of endogenous neural stem cells (NSCs) in rats following intracerebral hemorrhage (ICH) and the underlying mechanisms. Methods A total of 120 specific pathogen-free, adult, male Sprague-Dawley rats were included in this study. After establishment of ICH models in rats, PBS, Ad, or Ad-HIF-1α was administered via the ischemic ventricle. On the 1st, 7th, 14th, 21st and 28th d after ICH, rat neurological deficits were scored, doublecortin (DCX) expression in the subventricular zone cells was detected by immunohistochemical staining, and 5-bromo-2'-deoxyuridine (BrdU)-, BrdU/DCX-, and BrdU/glial fibrillary acidic protein-positive cells in the subventricular zone were counted using immumofluorescence method among PBS, Ad, and Ad-HIF-1α groups. Results On the 7th, 14th, 21st and 28th d after ICH, neurological deficit scores in the Ad-HIF-1α group were significantly lower than in the PBS and Ad groups (P<0.05). In the Ad-HIF-1α group, DCX expression was significantly increased on the 7th d, peaked on the 14th d, and then gradually decreased. In the Ad-HIF-1α group, BrdU-positive cells were significantly increased over time course, and significant difference in BrdU-positive cell counts was observed when compared with the PBS and Ad groups at each time point (P<0.01 or 0.05). On the 7th, 14th, 21st and 28th d after ICH, the number of DCX-, BrdU-, BrdU/DCX-, and BrdU/DCX-positive cells in the Ad-HIF-1α group was significantly greater than in the PBS and Ad groups (P<0.05). Conclusions HIF-1α gene can promote the proliferation, migration and differentiation of endogenous neural stem cells after ICH, thereby contributing to neurofunctional recovery after ICH.

771-775

Authors: GC Akuodor, AD Essien, E David-Oku, KC Chilaka, JL Akpan, B Ezeokpo, JOC Ezeonwumelu

Objective To evaluate the effect of aqueous leaf extract of Guiera senegalensis (G. senegalensis) on gastric mucosal damage using different ulcer models. Methods Considering the above claims, the present study was undertaken to validate the gastroprotective potential of the aqueous leaf extract of this plant against ethanol, water immersion and Aspirin induced ulcer models. Results The leaf extract (50, 100 and 200 mg/kg, p.o.) significantly (P<0.05) decreased the ulcer index in all assays used. Conclusions The results obtained, provide strong evidence of antiulcer activity of the leaf extract of G. senegalensis and support the traditional uses of the plant for the treatment of ulcer.

780-784

Authors: Herbert J Santos, Windell L Rivera

Objective To compare the sensitivity and specificity of direct fecal smear microscopy, culture, and polymerase chain reaction in the detection of Blastocystis sp. in human stool. Methods Human stool samples were collected from a community in San Isidro, Rodriguez, Rizal, Philippines. These samples were subjected to direct fecal smear microscopy, culture and polymerase chain reaction to detect the presence of Blastocystis sp. Results Of the 110 stool samples collected, 28 (25%) were detected positive for the presence of Blastocystis sp. by two or more tests. Culture method detected the highest number of Blastocystis-positive stool samples (n=36), followed by PCR of DNA extracted from culture (n=26), PCR of DNA extracted from stool (n=10), and direct fecal smear (n=9). Compared to culture, the sensitivity of the other detection methods were 66.7% for PCR from culture and 19.4% for both PCR from stool and direct fecal smear. Specificity of the methods was high, with PCR from culture and direct fecal smear having 97.3%, while PCR from stool at 95.9%. Conclusions In this study, in vitro culture is the best method for detecting Blastocystis sp. in human stool samples.

792-797

Authors: R Jafari, N Najafzadeh, MM Sedaghat, P Parvizi

Objective To assess molecular characterization, distribution, seasonal activities of sandfly species and Leishmania parasites infecting them for this zoonotic cutaneous leishmaniasis focus. Methods The collections were carried out in 2009–2011 using CDC traps, Sticky Papers and manual aspirator in and around the villages in Abarkouh district. Individual sandflies were characterized by PCR amplification and sequencing of fragments of their mitochondrial cytochrome b gene. Leishmania parasite infections within sandflies were performed by targeting Cyt b, ITS-rDNA, k-DNA and microsatellite genes. Results The PCR assays detected only Leishmania major (L. major). All infections (30) were found in the abundant and widespread vector Phlebotomus papatasi (P. papatasi). Small numbers of other sandfly species were also screened for infections, but none was found. Sergentomyia sintoni and P. papatasi were the predominant members in all locations of this district and in all habitats throughout the trapping season. Only five other sandfly species were found, namely Phlebotomus ansari, Phlebotomus caucasicus, Phlebotomus sergenti, Sergentomyia dentata and Sergentomyia merviney. Conclusions In the current survey, the only infections detected are of L. major in females of P. papatasi (30 out of 190). The rates of infection of P. papatasi by L. major are not significantly different in compare with other locations in Iran with no diversity of parasite strains. Zoonotic cutaneous leishmaniasis may have emerged only recently in Abarkouh district, and the reason may well be the instability of the transmission cycles there.

804-810

Authors: RS Renjith, AM Chikku, T Rajamohan

Objective To analyze the cytoprotective and antidiabetic activities as well as phytochemical composition of the immature inflorescence of Cocos nucifera belonging to the Arecaceae Family. Methods The phytochemical screening of inflorescence was done to determine the major constituents present in Cocos nucifera inflorescence. The free radical scavenging potential of inflorescence extracts were evaluated using in vitro radical scavenging assay models. Results The phytochemical analyses on inflorescence showed the presence of phenolic compounds, flavonoids, resins and alkaloids. The macronutrient analyses, on the other hand, showed the presence of carbohydrate, proteins and fibers. Administration of the methanol extract of coconut inflorescence to the diabetic rats showed dose dependent reduction in hyperglycemia. The cytoprotective property of coconut inflorescence was evidenced from the acute toxicological evaluation. The levels of serum aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase were significantly decreased in the diabetic rats treated with inflorescence when compared with the diabetic control rats. Conclusion The results obtained from the present study apparently proved the non-toxic nature and the cytoprotective and antihyperglycemic properties of coconut inflorescence.

817-822

Authors: Dan Liu, Si-Ping Dong, Guang-Ming Gao, Ming-Yu Fan, Zong-Jiu Zhang, Peng-Qian Fang

Objective To get scientific basis for further health education through the research of the road construction workers' KBP before and after the interventions of highway AIDS prevention project. Methods Multi-stage random sampling method was employeed to select workers of 8 sites from 14 sites along highway to investigate their AIDS knowledge, belief and performance (KBP) before and after highway AIDS prevention project. Results Over 90% of the investigated workers had ever heard about AIDS, and the non-skilled workers of lower educational level improved more after intervention. The correct answer rate of the three transmitting ways of AIDS of drivers which is the focused group of highway before and after intervention had the obvious statistical significance (P<0.05), and the other group's correct answer rates also had improved after intervention. Most people's understanding of preventing AIDS through correct use of condoms when having sex had a statistically significant difference(P<0.05) after prevention. The rates of using condoms of foremen and skilled workers when having sex with commercial sex worker/casual partner increased after intervention. Conclusions The health education of HIV among the road construction workers is effective and further health education of HIV prevention should be carried out among the road construction workers to improve their knowledge and awareness of avoiding the high-risk behaviors.

826-830

Authors: Bao Shan, Wang Li, Shu-Ying Yang, Zhuo-Ri Li

Objective To study the effect of estrogen on anovulatory dysfunctional uterine bleeding (ADUB). Methods Primary endometrial epithelial cells of Hainan Lizu female was cultured and hydrolytic activity of gelatinase was determined by gelatin zymography analysis. Cellular mRNA and protein synthesis was blocked respectively to determine whether the increased expression of MMP-2/9 was induced by estrogen. The expression of VEGF was blocked by siRNA. After treatment with various factors, MMP-9, VEGF, total Erk and phosphorylated Erk expression in primary uterine epithelial cells was detected by Western blotting analysis. Cell MMP-2/9mRNA levels was measured by real-time RT-PCR. Results The activity and expression of MMP2/9 was increased in the endometrium of patients with ADUB. Estrogen could up-regulate the expression of VEGF and activate Erk 1/2-Elk1 signal path. After interference by siRNA, ERK1/2 pathway was blocked in cells, and the expression of MMP-2/9 was down-regulated. ERK1/2 specific blocker U0126 blocked ERK phosphorylation, and it could down-regulate the expression of MMP-2/9. Conclusions The results showed that the estrogen can increase the expression of VEGF, and thus activate ERK1/2 pathway to induce MMP-2/9 expression.

835-838

Authors: Hua Wang, Wen-Jian Wang

Objective To investigate the relationship between CYP1A1 genetic polymorphisms and the invasion and metastasis of breast cancer. Methods The CYP1A1 gene polymorphism (an T-C transversion at nucleotide position 3801) was detected by the polymerase chain reaction and restriction fragment length polymorphism in 80 cases with breast cancer and 60 samples of normal breast tissue. The difference in genotypic distribution frequency between the groups, the correlation between the genotypes and the factors related to prognosis were analyzed. Results The incidence of homozygous and variant genotypes had no difference between the breast cancer group and controls group (P=0.746). The proportion of variant genotype increased as clinical stage (P=0.006) advanced, as well as with increased numbers of lymph node metastases (P=0.010). Conclusions In patients with breast cancer there is a correlation between the CYP1A1 CC allele and some factors indicating poor prognosis, including more lymph node metastases as well as a more advanced clinical stage.

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.