Asian Pacific Journal of Tropical Medicine

673-679

Authors: Satyajit Tripathy, Somenath Roy

Malaria is uncontrolled burden in the world till now. Despite of different efforts to develop antimalarial drug for decades, any anti-malarial drug can able to eradicate completely till now. Many anti-malarial substances are practically ineffectual because of their physicochemical limitations, cytotoxicity, chemical instability and degradation, and limited activities against intracellular parasites. Taking into consideration, the amount of research is going to conduct in the field of nanoparticle based drug delivery systems, lead to new ways of improving the treatment of infectious diseases. The study has focused on the progress and advancement of research on nanotechnology based drug delivery to eradicate the malaria. We like to focus the efficacy of nanotechnology based drug application for the opening out of novel chemotherapeutics in laboratory research, which may show the way to better use with age-old antimalarial drug and may draw the attention of pharmaceutical industries for the improvement and designing of effective anti-malarial drugs in future.

685-692

Authors: Khaled M.M. Koriem, Razmy E. Shahabudin, Rafiq Z. Jamaludin

Objective To evaluate a protective effect of Aristolochia gehrtii (A. gehrtii) leaves to inhibit liver toxicity and apoptosis in Schistosoma malayensis (S. malayensis) infection. Methods Forty male albino mice were divided into four equal groups: group 1 control including non-infected healthy mice and groups 2, 3 & 4 subcutaneously infected with S. malayensis cercariae where groups 3 & 4 pretreated with A. gehrtii leaves (200 mg/kg, bwt) & cinnamoylamide (250 mg/kg, bwt), respectively. Results S. malayensis caused a significant increase in serum AST, ALT, ALP, MDA, NO, bilirubin, urea, creatinine, total cholesterol, LDL, triglycerides, and HDL levels. The pretreatment of A. gehrtii leaves and cinnamoylamide significantly inhibited that increase. On the other hand, S. malayensis induced a significant decrease in serum total protein, albumin, globulin, albumin/globulin ratio, blood SOD and GPx, while A. gehrtii leaves and cinnamoylamide pretreatment increased the above parameters. Treatment with A. gehrtii leaves and cinnamoylamide to S. malayensis infected mice increased p53 expression but decreased bcl-2 expression. These results were supported by histopathological investigations. Conclusions A. gehrtii inhibits liver toxicity and apoptosis in S. malayensis infection and this effect is associated with the major cinnamoylamide ingredient of A. gehrtii leaves.

699-702

Authors: Hidayat Hussain, Ines Kock, Ahmed Al-Harrasi, Ahmed Al-Rawahi, Ghulam Abbas, Ivan R. Green, Afzal Shah, Amin Badshah, Muhammad Saleem, Siegfried Draeger, Barbara Schulz, Karsten Krohn

Objective To evaluate the antimicrobial potential of different extracts of the endophytic fungus Phoma sp. and the tentative identification of their active constituents. Methods The extract and compounds were screened for antimicrobial activity using the Agar Well Diffusion Method. Four compounds were purified using column chromatography and their structures were assigned using 1H and 13C NMR spectra, DEPT, 2D COSY, HMQC and HMBC experiments. Results The ethyl acetate fraction of Phoma sp. showed good antifungal, antibacterial, and algicidal properties. One new dihydrofuran derivative, named phomafuranol (1), together with three known compounds, phomalacton (2), (3R)-5-hydroxymellein (3) and emodin (4) were isolated from the ethyl acetate fraction of Phoma sp. Preliminary studies indicated that phomalacton (2) displayed strong antibacterial, good antifungal and antialgal activities. Similarly (3R)-5-hydroxymellein (3) and emodin (4) showed good antifungal, antibacterial and algicidal properties. Conclusions Antimicrobial activities of the ethyl acetate fraction of the endophytic fungus Phoma sp. and isolated compounds clearly demonstrate that Phoma sp. and its active compounds represent a great potential for the food, cosmetic and pharmaceutical industries.

709-714

Authors: Kittiyod Poovorawan, Salyavit Jitmitrapab, Sombat Treeprasertsuk, Thanunrat Thongmee, Apiradee Theamboonlers, Pisit Tangkijvanich, Piyawat Komolmit, Yong Poovorawan

Objective To report clinical outcomes and viral genotypes of acute symptomatic hepatitis E virus (HEV) infection in Thailand. Methods Forty patients with acute symptomatic HEV infection were recruited during 2009-2013. Clinical, demographic and laboratory data were collected. Diagnosis was accomplished by detection of anti-HEV IgM and/or HEV RNA in the serum or stool. HEV genotypes were classified by direct sequencing of RT-PCR products and phylogenetic analysis. Results The high risk group, comprising immune-compromised, liver cirrhosis and very elderly (>80 years) patients (17 cases), had higher levels of serum alkaline phosphatase at presentation compared with the low risk group. Two fatal cases resulted from acute hepatitis E in the high risk group. Initial clinical presentation did not show statistically significant differences. In six cases (6/40), the virus could be detected in serum or stool by RT-PCR and sequencing. Upon molecular characterization, the viruses were classified as HEV genotype 3f and were in the same cluster as Thai swine HEV. Conclusions Our data showed that acute HEV infection has various clinical presentations and outcomes. Higher levels of serum alkaline phosphatase were observed in high risk patients. All isolated viruses were identified as HEV genotype 3f possibly originating from swine.

720-724

Authors: Qiu-Ran Xu, Xin Liu, Ying-Min Yao, Qing-Guang Liu

Objective To investigate the expression of HSP90 and HIF-1α in human colorectal cancer tissue, the influence of HSP90 and HIF-1α on human colorectal cancer biological behavior and their related factors. Methods The expression of HSP90 and HIF-1α protein in human colorectal cancer as well as normal tissue were detected by immunohistochemical method. Results The positive expression rates of HSP90 and HIF-1α protein in normal human colorectal tissue as well as colorectal cancer tissue were 30% vs. 63.0%, 15.0% vs. 71.7%, respectively. There were significant difference (P=0.035 and P=0.005 respectively). The expression of HSP90 was significantly correlated with the differentiation, Dukes stages and lymph node metastasis (P<0.05), while the expression of HIF-1α was significantly correlated with the Dukes stages and lymph node metastasis (P<0.05). Association analysis showed that the expression of HSP90 protein was significantly correlated with that of HIF-1α protein(P<0.01). Conclusions The expression of HSP90 and HIF-1α protein may be related to the development, metastasis and invasion of human colorectal cancer; and their synergistic effects may participate in the development of the colorectal carcinoma.

730-734

Authors: Chun-Lai Zhang, Yue-Hong Li, Hong-Xia Zhou, Yu-Xin Zhang, Yong-Sheng Wang, Zhi-Yong Zhang, Ling-Li Meng, Xiao-Ming Shang

Objective To explore the protective effects of ginseng mixture on myocardial fibrosis (MF) in rats. Methods A total of 60 Wistar rats were randomly divided into control group without modeling operation, and another 4 groups using subcutaneous injections of isopropyl adrenaline for 10 d to set up the MF model: model group with saline lavage treatment after modeling, captopril group with captopril lavage, ginseng mixture group A and group B with low and high dose mixture treatment respectively. After treatment for 14 d, abdominal aorta and myocardial tissue were extracted to observe the pathological morphological changes and heart weight index in each group. Results The left ventricular weight and heart heavy index of captopril group and group B were significantly lower than that of model group and group A (P<0.05); Model group and group A showed a higher hydroxyproline (Hyp) content in myocardial tissue than the control group and lower catalase (CAT) activity than control group (P<0.05); captopril group and group B showed a lower Hyp content and higher CAT activity compared with group A and model group (P<0.05), a significantly lower level of serum glutathione peroxidase (GSH-PX) and CAT and a higher level of serum creatine kinase, lactate dehydrogenase and H2O2 in model group and group A were observed compared with the control group (P<0.05). A higher level of GSH-PX and CAT and a lower level of creatine kinase, lactate dehydrogenase and H2O2 in captopril group and group B were observed compared with group A and model group (P<0.05); and histopathological examination showed that in captopril group and group B, secretion of collagen fiber was significantly inhibited and myocardial injury was significantly lighter than that of model group. Conclusions Ginseng mixture plays a protective effect on myocardium by inhibiting antioxidant process of MF.

739-743

Authors: Ming-Lei Lang, Ai-Lin Qin, Jian-Min Li, Peng Fu

Objective To investigate the protective effect and mechanism of lithium chloride pretreatment on myocardial ischemia-reperfusion injury (I-RI) in rats. Methods A total of 60 SD rats were randomly divided into control group, model group, lithium chloride intervention group and L-arginine methyl ester + lithium chloride intervention group with 15 in each. The I-RI model was established in model group, the lithium chloride intervention group and L-arginine methyl ester + lithium chloride intervention group by method of seaming along left anterior descending coronary artery myocardial, control group was only opened the chest without seaming, ST-elevation within 2 min was regarded as modeling success. Model group did not adopted any intervention, lithium chloride intervention group was treated with lithium chloride injection 15 mg/kg by jugular venipuncture preoperatively, L-arginine methyl ester + lithium chloride intervention group was treated with intraperitoneal injection of 30 mg•kg−1•d−1 L-arginine methyl ester 7 d before the test, and intravenous catheter of 15 mg/kg lithium chloride preoperatively. The hydroxybutyric acid dehydrogenase (HBDH), creatine kinase isoenzyme (CK-MB), superoxide dismutase (SOD), malondialdehyde (MDA) level and nitric oxide synthase (NOS) activites were tested. Each large area of myocardial ischemia tissue was extracted for determination of the MDA content, SOD activity in tissue and serum, and morphological changes of myocardial tissue. Results SOD activity was highest in lithium chloride intervention group, followed by L-arginine methyl ester + lithium chloride intervention group, control group and model group (P<0.05); SOD activity was highest in L-arginine methyl ester + lithium chloride intervention group intervention group, followed by lithium chloride intervention group, control group and model group (P<0.05). MDA content of myocardial tissue was highest in model group, followed by L-arginine methyl ester + lithium chloride intervention group, the lithium chloride intervention group and control group (P<0.05); serum MDA content was highest in L-arginine methyl ester +lithium chloride intervention group, followed by model group, lithium chloride intervention group and control group (P<0.05). Compared with the control group, serum NOS was significantly higher in model group, lithium chloride intervention group and L-arginine methyl ester + lithium chloride intervention group (P<0.05), there was no statistical difference of serum NOS activity between the three groups (P>0.05); HBDH and CK-MB of plasma were highest in model group, followed by L-arginine methyl ester + lithium chloride intervention group, lithium chloride intervention group and control group (P<0.05). A significantly lighter myocardial damage was observed microscopically in lithium chloride intervention group than that in L-arginine methyl ester + lithium chloride intervention group and model group. Conclusions lithium chloride pretreatment can significantly reduce the myocardial I-RI, maintain structure and function of myocardial cells, improve the antioxidant ability of myocardial tissue, play an effective role on protecting myocardial I-RI.

749-753

Authors: Shu-Jie Zhao, Shi-Ji Wang, Hong-Xiang Li, Li-Hua Dong, Huai-Jiang He, Zhong-Min Liu, Yu-Shan Wang

Objective To tinvestigate the protective effect and mechanism of emodin pretreatment on intestinal mucosa of rats with intestinal ischemia-reperfusion injury. Methods A total of 50 SD rats were randomly divided into control group, model group, emodin groups of low, medium and high dose, with 10 in each group. Ischemia-reperfusion injury (I-RI) mode was established by using noninvasive clamp on superior mesentericartery (SMA). Control group and model group were pretreated with 0.5% sodium carboxymethyl cellulose solution lavage 2 h before operation, emodin groups of low, medium and high dose were given emodin lavage with 20, 40, 60 mg/kg pretreatment, femoral venous blood before the lavage pretreatment (T0) and 1 h ischemia (T1), and inferior vena venous blood after 1 h of reperfusion (T2) were extracted from each group of rats for detection of serum level of intestinal fatty acid binding protein (I-FABP), tumor necrosis factor (TNF-α), endotoxin, interleukin 6 (IL-6), and the content of diamine oxidase (DAO); After model establishment, the rats were sacrificed, intestine homogenate was prepared by using blind intestinal tissue to detect intestinal tissue myeloperoxidase (MPO), malondialdehyde (MDA) and superoxide dismutase (SOD) levels. And upper small intestine tissue was retrieved, followed by fixation and conventional HE staining to observe intestinal tissue morphology under light microscopy. Results In emodin groups of low, medium and high dose at T1 and T2, I-FABP, TNF-α, endotoxin, IL-6 and DAO level were significantly lower than that of model group (P<0.05); in emodin group of low, medium and high dose, MPO and MDA content in intestinal tissue homogenate was significantly lower than that in model group (P<0.05), SOD level was significantly higher than that of model group (P<0.05). Intestinal damage of emodin low, medium and high dose groups were significantly lighter than model group. Conclusions Emodin pretreatment has certain protective effect on intestinal mucosa in ischemia reperfusion injury.

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.