Asian Pacific Journal of Tropical Medicine

253-259

Authors: Santram Lodhi, Abhay K Singhai

Objective To evaluate wound healing potential of flavonoid fractions of Martynia annua (M. annua) Linn. leaves in diabetic rats on the basis of folkloric information and preliminary study. Methods The flavonoid compound luteolin and apigenin were isolated from dried leaves of plant by column chromatography. The two concentrations (0.2% and 0.5% w/w) of luteolin and flavonoid fraction were selected for topically applied as ointment on diabetic wound. The Povidone Iodine Ointment USP was used as a reference. On 18th days, protein content, hydroxyproline and antioxidants (SOD, CAT and GSH) level in granuloma tissues were determined. Results The results showed that, percent wound contraction were observed significantly (P<0.01) greater in MAF fraction and 0.5% w/w of luteolin treatment groups. Presence of matured collagen fibres and fibroblasts with better angiogenesis were observed in histopathological studies. Conclusions In conclusion, our findings suggest that flavonoid fraction (MAF) and luteolin (0.5% w/w) may have potential benefit in enhancing wound healing in diabetic condition, possibly due to free-radical scavenging activity of plant.

265-272

Authors: Ya-Fei Zhu, Guo-Lan Gao, Sheng-Bo Tang, Zhen-Dong Zhang, Qing-Shui Huang

Objective To investigate effect and possible mechanisms of silencing human WFDC2 (HE4) gene on biological behavior changes as cell proliferation, apoptosis, movement and invasion of human serous ovarian cancer cell line SKOV3. Methods Lentiviral WFDC2 gene sequence of small interfering siRNA was stablely transfected into SKOV3 identified by Q-PCR and western-blot. Obtained SKOV3 stable strains with silenced HE4 were measured by proliferation, apoptosis, migration, and invasion. Results Gene sequencing showed that the oligonucleotides were successfully inserted into the expected site. After silencing HE4 in the SKOV3, proliferation was significantly inhibited (P<0.05). G0/G1 phase was arrested by the cell cycle (P<0.01) and capacity of the migration and invasion decreased significantly (P<0.01). Slight early apoptosis ratio and no change of late apoptosis were found without change of Caspase-3 or Bcl-2 protein. Proteins involed in ERK pathway as phosphorylated protein as p-EGFR, p-ERK decreased and protease protein involved in tissue remoding as matrix metalloproteinases MMP-9, MMP-2 and cathepsin B decreased compared with control group. Conclusions HE4 gene plays an important role in regulating proliferation, apoptosis, migration, invasion of serous ovarian cancer cells by ERK pathway and protease system. Its role in apoptosis needs to be further explored, and it may be a potential target for serous ovarian cancer.

280-284

Authors: Sunil Mistry, KR Dutt, J Jena

Objective To investigate the hepatoprotective potential of Sida cordata (Malvaceae) (S. cordata) in experimental rats to validate its traditional claim. Methods Wister albino rats were divided into 6 groups: Group I served as control; Group II served as hepatotoxic (CCl4 treated) group; Group III, IV and V served as (100, 200 and 400 mg/kg b.w.) S. cordata leaf extract (SCLE) treated groups; Group VI served as positive control (Silymarin) treated group. Liver marker enzymes serum glutamate oxyloacetic transaminase, serum glutamic pyruvic transaminase, pancreatic enzymatic antioxidants superoxide dismutase (SOD), lipid peroxidation, catalase (CAT), reduced glutathione (GSH) were measured and compared along with histopathological studies. Results Obtained results show that the treatment with SCLE significantly (P<0.05-<0.001) and dose-dependently reduced CCl4 induced elevated serum level of hepatic enzymes. Furthermore, SCLE significantly (up to P<0.001) reduced the lipid peroxidation in the liver tissue and restored activities of defence antioxidant enzymes GSH, SOD and CAT towards normal levels, which was confirmed by the histopathological studies. Conclusions The results of this study strongly indicate the protective effect of SCLE against CCl4 induced acute liver toxicity in rats and thereby scientifically support its traditional use.

296-299

Authors: Hong Wang

Objective To investigate the effect of ophiopogon-polysaccharide on adiponectin (APN) in gestational diabetic mellitus rat. Methods The model of gestational diabeties mellitus rat were established with streptozotocin by intraperitoneal injection. Model rats were randomly divided into ophiopogon-polysaccharide 125 mg/kg group, 250 mg/kg group, 500 mg/kg group, GDM group and normal pregnancy group (n=10). Ophiopogon-polysaccharide was given by intragastric administration for 14 days. The fasting blood glucose levels of rats at the 0, 7, 14 day were detected respectively. The serum insulin level and APN in fat tissue and placenta were detected at the 14 day. Results The fasting blood glucose level, serum insulin level and APN mRNA in fat tissue and placenta were significantly decreased in gestational diabetes rats treated by ophiopogon-polysaccharide compared with model group (P < 0.05). Conclusions Ophiopogon-polysaccharide could decrease the fasting blood glucose level and serum insulin level, and improve APN mRNA in fat tissue and placenta.

305-310

Authors: Zhen-Guo Cheng, Guo-Dong Zhang, Peng-Qiang Shi, Bao-Shun Du

Objective To explore the expression of Nrf2/ARE pathway in hindbrain tissue after the traumatic brain injury (TBI) and its anti-oxidative stress effect in the secondary nerve injury. Methods The mice with Nrf2 gene knockout were used for the establishment of brain injury model. The experimental animals were divided into four groups: (Nrf2+/+) sham-operation group, (Nrf2+/+) brain injury group, (Nrf2−/−) sham-operation group and (Nrf2−/−) brain injury group. The specimen 24 h after cerebral trauma was selected. Then RT-PCR method was adopted to detect the expression of Nrf2 mRNA in brain; Western blotting method was adopted to detect the levels of Nrf2, HO-1 and NQO1 proteins in brain; ELISA method was adopted to detect the oxidative stress indicators: protein carbonyls, 4-hydroxy-2-nonenal (4-HNE) and 8-hydroxy-2′-deoxyguanosine (8-OHdG). Results The Nrf2 mRNA and protein of Nrf2−/− mice were not expressed, and the difference of the relative amount of Nrf2 mRNA between Nrf2+/+ TBI group and Nrf2+/+ sham-operation group was not statistically significant (P>0.05); the level of Nrf2 protein in Nrf2+/+ TBI group increased significantly compared with the Nrf2+/+ sham-operation group (P<0.01); in the sham-operation groups, the levels of HO-1 and NQO1 proteins of Nrf2−/− mice decreased obviously compared with the Nrf2+/+ mice (P<0.01); after brain injury, the levels of HO-1 and NQO1 proteins of Nrf2+/+ mice increased obviously compared with the corresponding sham-operation group (P<0.01); the levels of HO-1 and NQO1 proteins of Nrf2−/− mice in TBI group had no obvious change compared with the corresponding sham-operation group (P>0.05); there was only a little amount of expression of protein carbonyls, 4-HNE and 8-OHdG proteins in brain tissues in the Nrf2+/+ and Nrf2−/− sham-operation groups, and the difference was not statistically significant (P>0.05); after brain injury, the three oxidative stress indicators were significantly up-regulated in the Nrf2+/+ and Nrf2−/− groups, and the up-regulation of the latter group was more significant (P<0.01). Conclusions After TBI the Nrf2/ARE pathway is activated and the activity of Nrf2 transcription regulation increases. However, the regulation dose not occur in the gene transcription level and only could increase the Nrf2 protein level, while the mRNA expression level has no obvious change. The nerve cell protective effect of Nrf2/ARE pathway in TBI achieves through inhibiting the oxidative stress injuries.

315-319

Authors: Samiran Bandyopadhyay, Pabitra H Patra, Achintya Mahanti, Dipak K Mondal, Premanshu Dandapat, Subhasis Bandyopadhyay, Indranil Samanta, Chandan Lodh, Asit K Bera, Debasish Bhattacharyya, Mihir Sarkar, Kishore K Baruah

Objective To evaluate the antimicrobial efficacy of berberine, a plant alkaloid. Methods Five multi-drug resistant (MDR) STEC/EPEC and five MDR ETEC isolates from yaks with haemorrhagic diarrhoea were selected for the study. Antibacterial activity of berberine was evaluated by broth dilution and disc diffusion methods. The binding kinetics of berberine to DNA and protein was also enumerated. Results For both categories of enterovirulent Escherichia coli (E. coli) isolates, berberine displayed the antibacterial effect in a dose dependent manner. The MIC50 of berberine chloride for STEC/EPEC isolates varied from 2.07 μM to 3.6 μM with a mean of (2.95 ± 0.33) μM where as for ETEC strains it varied from 1.75 to 1.96 μM with a mean of (1.87 ± 0.03) μM. Berberine bind more tightly with double helix DNA with Bmax and Kd of (24.68±2.62) and (357.8±57.8), respectively. Berberine reacted with protein in comparatively loose manner with Bmax and Kd of (18.9±3.83) and (286.2±113.6), respectively. Conclusions The results indicate clearly that berberine may serve as a good antibacterial against multi drug resistant E. coli.

325-332

Authors: Loganathan Karthik, Gaurav Kumar, Kokati Venkata Bhaskara Rao

Objective To investigate the antioxidant activity of marine actinobacteria. Methods The content of total phenolics, the level of antioxidant potential by DPPH radical scavenging activity, metal chelating activity, FRAP method, β carotene assay and NO scavenging activity in extract were determined. Results In all the methods the extract exhibited good scavenging activity except NO scavenging activity. The IC50 values of marine actinobacteria extract on DPPH radical were found to be 41.09 μg/mL. The zone of color retention was 12 mm in β-carotene bleaching assay. DNA protective efficiency of the extracts was also studied using UV-photolysed H2O2-driven oxidative damage to pBR322. HPLC analysis identified some of the major phenolic compounds in extracts, which might be responsible for the antioxidant potential and cyto-protection. It showed a 100% cytotoxic effect in brine shrimp lethality assay within 10 mins. The novel actinobacteria was identified as Streptomyces LK-3 (JF710608) through 16S rDNA Sequencing. Conclusions The results obtained suggest that the extracts bear anti-cancer metabolites and could be considered as a potential source for anti-cancer drug development.

Ya-Ping Li

Authors: 220-223

Objective To investigate the expression and potential roles of G protein-coupled receptor kinase 6 (GRK6) in hepatocellular carcinoma (HCC) patients. Methods Immunohistochemistry and Western blot was performed to determine GRK6 expression in 73 HCC samples. And the correlation with clinicopathological features was also analyzed. Results GRK6 expression was significantly higher in HCC than that in normal hepatic tissue. GRK6 was positively correlated with proliferation marker Ki-67, clinical stage, metastasis and survival time. Conclusions Our results suggested that GRK6 overexpression plays an important role in HCC. Monitoring the expression of GRK6 maybe helpful in early diagnosis and prognosis of HCC.

176-179

Authors: G Renugadevi, T Ramanathan, priya R Shanmuga, P Thirunavukkarasu

Objective To investigate the studies on effects of Andrographis paniculata (A. paniculata) (Burm.f.) and Andrographis lineata (A. lineata) nees (Family: Acanthaceae) extracts against two mosquitoes Culex quinquefasciatus (Cx. quinquefasciatus) (Say.) and Aedes aegypti (Ae. aegypti) (Linn.). Methods The aqueous and petroleum ether extracts of two plant species, A. paniculata and A. lineate were examined against the larvae of A. aegypti (L.) and Cx. quinquefasciatus with gradually increasing concentration ie. from 50 to 200 ppm of solvent extracts and to test their activity in combination with each other. Results In a 24 h bioassay experiment with plant extracts, highest mortalities were recorded at 200 ppm of concentrations for leaves of A. lineta and A. paniculata individually. For combination effect, only 150 ppm of the mixture of solvent extracts of petroleum ether: aqueous (1:1) extracts showed 100% mortality after 24 h of exposure. Conclusions The results show that, insecticides of plant combination is ecofriend and has better larvicidal activity compared to individual extracts.

188-194

Authors: Uma Chidambaram, Vanitha Pachamuthu, Suganya Natarajan, Bhakkiyalakshmi Elango, Suriyanarayanan, Kunga Mohan Ramkumar

Objective To determine the phenolic content in Codariocalyx motorius root extract and to evaluate its antioxidant properties using various in vitro assay systems. Methods The antioxidant activity was evaluated based on scavenging of 1,1-diphenyl-2-picrylhydrazyl, hydroxyl radicals, superoxide anions, nitric oxide, hydrogen peroxide, peroxynitrite, reducing power and by inhibition of lipid peroxidation which was estimated in terms of thiobarbituric acid reactive substances. Results The root extract of the Codariocalyx motorius (C. motorius) exhibited potent total antioxidant activity that increased with increasing amount of extract concentration, which was compared with standard drug such as quercetin, butylated hydroxytoluene, tocopherol at different concentrations. The different concentrations of the extracts showed inhibition on lipid peroxidation. In addition, the extracts had effective reducing power, free radical scavenging, super oxide anion scavenging, nitric oxide scavenging, lipid peroxidation, and total phenolic content depending on concentration. High correlation between total phenolic contents and scavenging potential of different reactive oxygen species (r2=0.831–0.978) indicated the polyphenols as the main antioxidants. Conclusions Codariocalyx motorius (C. motorius) root possess the highly active antioxidant substance which can be used for the treatment of oxidative stress-related diseases.

200-204

Authors: M. Tolulope Olaleye, Afolabi C. Akinmoladun, Olamide O. Crown, Katty E. Ahonsi, A.O. Adetuyi

Objective To investigate the restorative effect of Pterocarpus erinaceus (P. erinaceus) and homopterocarpin, an isoflavonoid isolated from it, on indomethacin-induced disruption in gastric homeostasis in rats. Methods Adult rats were divided into five groups and fasted for 48 h before treatment. Group 1 received olive oil (vehicle), group 2 received 25 mg/kg indomethacin while groups 3–5 received cimetidine (100 mg/kg), homopterocarpin (25 mg/kg) and P. erinaceus ethanolic stem bark extract (100 mg/kg) respectively. After 1 h, all the groups except group 2 were administered 25 mg/kg of indomethacin. One hour later, the rats were sacrificed and the ulcer index and other gastroprotective indices were evaluated. Results Indomethacin caused significant injury to the stomach of the rats as reflected in the ulcer indices (9.0±1.4) as compared with that of control (2.0±0.0). Equally, there were significant increases in gastric acid concentration and malondialdehyde level in the stomachs of the ulcerated animals compared with the control. However mucus content, reduced gluthatione level and gastric pH were significantly reduced in the ulcerated animals compared with the control. Pretreatment with either Pterocarpus bark extract or homopterocarpin reversed the effects of indomethacin on the evaluated parameters. Conclusions These results indicate that both homopterocarpin and Pterocarpus extract offered gastroprotection against indomethacin-induced ulcer by antioxidative mechanism and the modulation of gastric homeostasis. The results also suggest that homopterocarpin might be responsible for, or contribute to the antiulcerogenic property of P. erinaceus.

212-215

Authors: Dina Abdulla Muhammad Zaglool, Amr Mohamed, Yousif Abdul Wahid Khodari, Mian Usman Farooq

Objective To evaluate the validity of Crypto-Giardia antigen rapid test (CA-RT) in comparison with the conventional modified Ziehl-Neelsen acid fast (MZN-AF) staining method for the diagnosis of cryptosporidiosis. Methods Fifteen preserved stool samples from previously confirmed infections were used as positive controls and 40 stool samples from healthy people were used as negative control. A total of 85 stool samples were collected from suspected patients with cryptosporidiosis over 6 months during the period from January till June, 2011. The study was conducted in the department of parasitology, central laboratory, Alnoor Specialist Hospital, Makkah, Saudi Arabia. All samples were subjected to CA-RT and conventional MZN-AF staining method. Validation parameters including sensitivity (SN), specificity (SP), accuracy index (AI), positive predictive value (PPV), and negative predictive value (NPV) were evaluated for both tests. Results Out of 15 positive controls, CA-RT detected 13 (86.7%) while MZN-AF detected 11(73.3%) positive cases. However, CA-RT detected no positive case in 40 normal controls but MZN-AF detected 2(5%) as positive cases. Based on the results, the SN, SP, AI, PPV and NPV were high in CA-RT than MZN-AF staining method, ie., 86.7% vs. 73.3%, 100% vs. 95%, 96.4% vs. 89.1%, 100% vs. 84.6% and 95.2% vs. 90.5%, respectively. Out of a total of 85 suspected specimens, CA-RT detected 7(8.2%) but MZN-AF detected 6(7.1%) cases as positive. Conclusions CA-RT immunoassay is more valid and reliable than MZN-AF staining method.

224-227

Authors: Jun Zhu

Objective To probe into the influences of Chinese herbal compound on the growth and metabolism of weight-bearing bones of tail-suspended rats. Methods Twenty-four male SD rats were randomly divided into blank control group (eight), tail-suspended control group (eight) and Chinese medicine treatment group (eight) according to their weights. No treatment was done for the blank control group. Double distilled water lavage was performed daily for the tail-suspended control group. On the basis of the tail-suspended rat model, the rats were given Chinese herbal compound lavage every day in Chinese medicine treatment group. This compound includes mulberry, Poria cocos and barbary wolfberry, etc. The test cycle was four weeks. The rats were killed after the experiment. The right femoral bone was taken out for the physical measurements, and the left femoral bone was for the three-point bending test. The influences of Chinese herbal compound on femoral bone growth and biomechanical properties of simulated weightlessness rats were observed. Results (1) After simulated weightlessness (tail-suspension), compared with the blank control group, all the physiological indexes of rat femoral bone decreased in tail-suspended group and Chinese medicine treatment group (P<0.05). The strength and rigidity of rat femoral bone decreased in tail-suspended group (P<0.01). The maximum load and rigidity coefficient also decreased with the increasing toughness coefficient in the control group (P<0.01). (2) After the countermeasure of Chinese herbal compound, each biomechanical indexes showed the tendency of increasing in Chinese medicine treatment group, and theses indexes were close to those of the blank control group (P<0.05), which indicated that the bone loss caused by simulated weightlessness was improved. Conclusions Chinese herbal compound for tonifying kidney could effectively prevent the bone loss and have some enhancements on the bone biomechanical properties.

232-236

Authors: Yu-Hong Xia, Li Yao, Zhan-Xin Zhang

Objective To explore the relationship between IL-1β, IL-1Ra gene polymorphism and the occurrence of polycystic ovary syndrome (PCOS) infertility. Methods A total of 59 PCOS infertility cases visiting the reproductive center of our hospital from Mar. 2010 to Mar. 2012 and 56 healthy women were selected. ELISA method was used for the detection of IL-1β, IL-1Ra levels, and the levels of serum supersensitivity C reaction protein (US-CRP), insulin (FINS), follicule-stimulating hormone (FSH) and fasting blood-glucose (FBG) were detected. PCR analysis technology was adopted to detect the gene polymorphism of the 511 site of IL-1β and the second introne of IL-1Ra. Results The levels of IL-1β, IL-1Ra, US-CRP, FINS and FBG in blood serum of patients in PCOS group were significantly higher than those in control group (P<0.05 or P<0.01). The level of FSH in PCOS group was significantly lower than that in control group (P<0.05). The genotypic frequency of T/T, the 511 site of IL-1β in PCOS group was 42.37%, significantly higher than 12.50% in control group (P<0.01). The frequency of T allele was also significantly higher than that in control group (P<0.01). The genotypic frequency of I/V, the second introne of IL-1Ra in PCOS group was 20.34%, signiciantly higher than 3.57% in control group (P<0.05). The frequency of V allele in PCOS group was significantly higher than that in control group (P<0.05). Conclusions T allele of the 511 site of IL-1β gene and V allele of the second introne of IL-1Ra gene might be the genetic basis of the rising of IL-1β, IL-1Ra and US-CRP levels in blood serum of PCOS patients, and are associated with the infertility occurrence of PCOS patients.

242-245

Authors: Yu-Cai Zhang

Objective To explore a rapid diagnostic method in neonatal sepsis and bacterial meningitis. Methods The primers were designed and synthesized based on 16S rRNA gene of Staphylococcus aureus. Four specimens of Staphylococcus aureus, 16 specimens of coagulase-negative Staphylococci, 2 specimens of Enterococci, 3 specimens of Streptococcus, 1 specimen of Micrococcus, 3 specimens of Escherichia coli, 4 specimens of Klebsiella pneumoniae, 3 specimens of Pseudomonas aeruginosa, 2 specimens of Enterobacter cloacae, and 5 specimens of Acinetobacter were tested by loop-mediated isothermal amplification (LAMP) assay. A total of 118 clinical specimens of sepsis and non-sepsis were collected and detected with both LAMP assay and blood culture. Results By designing primers specific for Staphylococcus aureus, specimens containing different kinds of pathogens were carried out by LAMP assay, and our data showed LAMP technology for the specific detection of Staphylococcus aureus in samples was successfully established. All clinical specimens of sepsis and non-sepsis were tested by both blood cultures and LAMP, and our data showed that compared wit blood culture method, the LAMP technology showed significantly high detection rate (P <0.01). Conclusions As a quick and easy detection of Staphylococcus aureus, the LAMP technology was successfully established, laid the foundation for the diagnosis and treatment of children Staphylococcus aureus sepsis, and showed great promotion and application value.

249-250

Authors: Sunny Garg, Richik Tripathi, Kamlakar Tripathi

Leishmaniasis affects both the visceral and cutaneous tissues in body. Oral Mucosal involvement in leishmaniasis is rare and is often overlooked. We present a case 17 year old boy from the north east region of Bihar who has a history of visceral leishmaniasis one year back, came to the department of oral surgery for treatment of persistent oral ulcers. Oral examination did not give any diagnostic information while systemic examination revealed enlarged spleen and low grade fever. Patient was screened for leishmaniasis by rK39 based immunochromatographic strip test which came to be positive. Biopsy of the ulcer as well as splenic and bone marrow aspirate confirmed the presence of leishmaniasis. Patient was administered Amphotericin B for 20 days following which significant clinical and haematological improvement followed.

85-94

Authors: Wing-Chui Chong, Rusliza Basir, Yam Mun Fei

Malaria is an intra-cellular parasitic protozoon responsible for millions of deaths annually. Host and parasite genetic factors are crucial in affecting susceptibility to malaria and progression of the disease. Recent increased deployment of vector controls and new artemisinin combination therapies have dramatically reduced the mortality and morbidity of malaria worldwide. However, the gradual emergence of parasite and mosquito resistance has raised alarm regarding the effectiveness of current artemisinin-based therapies. In this review, mechanisms of anti-malarial drug resistance in the Plasmodium parasite and new genetically engineered tools of research priorities are discussed. The complexity of the parasite lifecycle demands novel interventions to achieve global eradication. However, turning laboratory discovered transgenic interventions into functional products entails multiple experimental phases in addition to ethical and safety hurdles. Uncertainty over the regulatory status and public acceptance further discourage the implementation of genetically modified organisms.

102-109

Authors: C. Panneerselvam, K. Murugan, K. Kovendan, P. Mahesh Kumar, J. Subramaniam

Objective To explore the larvicidal and pupicidal activity of Euphorbia hirta (E. hirta) leaf extract and Bacillus sphaericus (B. sphaericus) against the malarial vector, Anopheles stephensi (An. stephensi). Methods The larvicidal and pupicidal activity was assayed against An. stephensi at various concentrations ranging from (75-375 ppm) under the laboratory as well as field conditions. The LC50 and LC90 value of the E. hirta leaf extract was determined by probit analysis. Results The plant extract showed larvicidal effects after 24 h of exposure; however, the highest larval mortality was found in the methanol extract of E. hirta against the first to fourth instars larvae and pupae of values LC50= 137.40, 172.65, 217.81, 269.37 and 332.39 ppm; B. sphaericus against the first to fourth instars larvae and pupae of values LC50= 44.29, 55.83, 68.51, 82.19 and 95.55 ppm, respectively. Moreover, combined treatment of values of LC50= 79.13, 80.42, 86.01, 93.00 and 98.12 ppm, respectively. No mortality was observed in the control. Conclusions These results suggest methanol leaf extracts of E. hirta and B. sphaericus have potential to be used as an ideal eco-friendly approach for the control of the malarial vector, An. stephensi as target species of vector control programs. This study provides the first report on the combined mosquito larvicidal and pupicidal activity of this plant crude extract and bacterial toxin against An. stephensi mosquitoes.

115-118

Authors: Fooladi AA Imani, Islamieh D Iman, Doust R Hosseini, A Karami, SM Marashi

Objective To study virulence and regulatory genes (hlyA, ctxB, tcpI) in clinical strains of Vibrio cholerae (V. cholerae), simultaneously. Methods Three important genes, tcpI, hlyA and ctxB were used for detection of toxigenic and pathogenic V. cholera by chain reaction assay method. Results According to the results of the PCR, the incidence of hlyA, tcpI, and ctxB genes in clinical isolates was obtained as 94.7% (72 sample), 90.8% (69 sample), and 92.1% (70 sample), respectively. Five strains possessed all genes except ctxB, six strains possessed all genes except tcpI, four strains possessed all genes except hlyA, one strain possessed only hlyA and 60 strains contained a combination of three genes, Including hlyA, ctxB and tcpI. Conclusions Result show that this method could be reliable to detect toxigenic-pathogenic strains of V. cholerae in Iran.

126-130

Authors: Caroline E. Omoti, Chiedozie K. Ojide, Patrick V. Lofor, Emeka Eze, Joy C Eze

Objective To investigate the malaria parasitemia, CD4+ cell counts and some haematological indices among HIV-malaria co-infected adult patients with highly active antiretroviral therapy (HAART). Methods A total of 342 adult HIV positive subjects were recruited at the consultant outpatient HIV/AIDS clinic, University of Benin Teaching Hospital, Benin City, Nigeria between June 2011 to November 2011. Blood samples were taken for malaria parasite count, CD4+ cell count and other haematological counts. Results Out of the 342 adult HIV positive subjects a total of 254 patients (74.3%) were found to have malaria parasitemia. The incidence of malaria parasitemia increased with advancing clinical stage of HIV infection and this was statistically significant (P=0.002). There was no statistical significance when gender was compared with the HIV-malaria status (P >0.05). Of the 254 co-infected patients, 134 (52.8%) had high parasitemia (>1.25×109/L). Sixty patients were found to be hyperparasitemic (>2.5 parasites/L). There was a significant association between CD4+ cell count and having significant parasitemia (P < 0.000 1). About half (50.8%) of co-infected patients had CD4+ cell count ⩽ 200/μL, and majority (44.9%) of this population also had significant parasitemia. Anaemia and thrombocytopenia were not significantly associated with HIV-malaria co-infection (P > 0.05). Conclusions The prevalence of parasitemia is high among the HIV/AIDS infected patients.

135-138

Authors: Guadalupe García-Elorriaga, Mayté Martínez-Velázquez, Verónica Gaona-Flores, Guillermo del Rey-Pineda, César González-Bonilla

Objective To assess the usefulness of IGRA test (QuantiFERON®-Cell mediated immune) compared with the tuberculin skin test. Methods A cross-sectional study was carried out in Mexico, 25 infected patients with HIV-AIDS and the suspicion or with latent tuberculous infection (LTBI) who were >18 years of age and without treatment for tuberculosis (TB), were enrolled in the study. Results Median cluster of differentiation (CD4) count was 364 cells/μ L and median HIV viral load was 50 copies/mL. Overall, 20 patients (80%) had at least one positive diagnostic test for LTBI: four (16%) had a positive tuberculin skin test and 19 (76%), a positive QuantiFERON®-tuberculosis. Conclusions No agreement is found between the two diagnostic tests: k = −0.004, 95% confidence interval (−0.2219, 0.2210). Additional longitudinal studies among HIV-infected populations with high prevalence of TB are needed to further assess the usefulness of IGRAs in this patient population.

145-149

Authors: Li Zhang, Shu-Zhen Dai, Xiao-Dong Nie, Lei Zhu, Feng Xing, Li-Ya Wang

Objective To explore the effect of Salvia miltiorrhiza on diabetic retinopathy (DR). Methods Diabetic mice of natural incidence type with monogenic inheritance were selected. Alloxan was injected into the caudal vein of mice once to induce DR. The structural changes of retina tissue in normal mice, DR mice and mice with high, medium and low dose of Salvia miltiorrhiza injection were observed under microscope. Then the blood glucose concentration and malonaldehyde (MDA) content were detected. Results There were some microaneurysms in retina of DR group, number of gangliocyte was decreased significantly, and cells were sparse and in disorder. After modeling, the blood glucose level of high-dose Salvia miltiorrhiza group (SM III group) was significantly different from DR group (P<0.01). Till the tenth week, the blood glucose level of all SM groups was decreased significantly compared with DR group (P<0.01). The effective rates of three SM groups were 93.8%, 76.4% and 50.3%, respectively. After ten weeks, MDA content of DR group was significantly higher than those of the normal control group and SM group (P<0.01), and medium and low dose SM groups had significantly higher MDA than that of normal control group (P<0.01). Conclusions Salvia miltiorrhiza had certain protective effect on DR mice through the blood-ocular barrier.

153-155

Authors: Jin Ren, Li-Xuan Wang, Xue-Cheng Ji, Jia-Ying Zhou, Man-Ying Zhang

Objective To investigate ultrastructural changes in pulmonary tissue of a rat model of pulmonary fibrosis following treatment with compound Carapax trionycis (C. trionycis; Biejia in Chinese) formula. Methods Sixty male Sprague-Dawley rats were randomly divided into four groups (n=15): compound C. trionycis formula high-, middle-, and low-dose groups as well as model group. Pulmonary fibrosis was induced by bleomycin. Five rats from each group were sacrificed on day 7, 14 and 28 of the drug treatment, respectively. The pulmonary tissue was harvested followed by hematoxylin-eosin staining and subsequent transmission electron microscopy. The Szapiel's method was used to assess the degree of alveolitis and pulmonary fibrosis. Results Compared with the model group, the compound C. trionycis formula groups had slighter pulmonary alveolitis after the 7-day treatment and also had alleviated alveolar inflammation and pulmonary fibrosis after the 14-day treatment. After the 28-day treatment, the compound C. trionycis formula groups showed deposition of a small amount of fibrous tissue and lesions occupying less than 21% of the whole lung area, while the model group showed focal or diffuse fibrous deposition, narrow alveolar cavity, disordered lung structure, and lesions in larger than 51% of the whole lung area. Conclusions The compound C. trionycis formula can inhibit the proliferation of collagen fibers and resist pulmonary fibrosis.

162-166

Authors: Pin Ha, Xu-Qing Zhuge, Qian Zheng, Bing Shi, Cai-Xia Gong, Yan Wang

Objective To obtain descriptive information of behavioral pattern in Chinese school-aged children with cleft lip and palate. Methods A total of 93 cleft lip and palate patients between the age of 6-11 year-old and treated at West China Stomatology Hospital were selected. And another 100 unaffected controls, matched for age and gender, were recruited randomly from a common primary school in Chengdu. Chart review of medical records was used to obtain psychosocial checklists. Scores were compared with published norms and controls to evaluate the risk of problems, separately for three diagnostic groups. Results The patients group had lower scores of social and academic competencies, especially those with facial deformity or speech problem. No difference was found in the aspect of activity competency. All patients showed elevations in behavior problems. But the type of behavior problems varied in different genders. Conclusions Chinese school-aged children with cleft lip and palate are at raised risk for social and academic difficulties. Specific pattern of behavior problems displays differently depending on gender of the patient.

1-10

Authors: Fathima Stanley Rosarin, Vadivel Arulmozhi, Samuthira Nagarajan, Sankaran Mirunalini

Objective To synthesize silver nanoparticles by amla extract, screen the cytotoxic, oxidative stress and apoptotic effect of silver nanoparticles (AgNPs) on Hep2 cell line (laryngeal carcinoma cells) in vitro, and to compare the effect of Phyllanthus emblica (P. emblica) (amla) with AgNPs synthesized by amla and 5-FU. Methods AgNPs was synthesized by P. emblica (aqueous extract) and nanoparticles were characterized UV-Vis spec, the presence of biomoloecules of amla capped in AgNPs was found by FT-IR analysis, shape and size were examined by SEM and DLS. Cytotoxicity of experimental drugs was tested to find IC50 value. ROS generation in cells have been measured by DCFH-DA staining, AO-EtBr, Rhodamine-123 staining and DNA fragmentation were performed to assess apoptotic cell death, mitochondrial membrane potential and apoptotic DNA damage, respectively. Oxidative stress was analyzed by measuring lipid peroxides and antioxidants level to understand the cancer cell death by pro-oxidant mechanism. Results PE-AgNPs was synthesized and confirmed through kinetic behavior of NPs. The shape of PE-AgNPs was spherical and cubic since it was agglomerated, and the nanoparticle surface was complicated. Average particle size distribution of PE-AgNPs was found to be 188 nm. Potent biomolecules of P. emblica such as polyphenols were capped with AgNPs and reduced its toxicity. In cytotoxicity assay the concentration in which the maximum number of cell death was 60 μg/mL and 50 μg/mL for P. emblica (alone) and AgNPs, respectively and IC50 values were fixed as 30 μg/mL and 20 μg/mL. ROS generation, apoptotic morphological changes, mitochondrial depolarization, DNA damage and oxidative stress was observed as more in AgNPs treated cells than in P. emblica (30 μg/mL) (alone) treated cells and 5-FU treated cells gave similar result. Conclusions The results suggest that the AgNPs are capped with biomolecules of amla enhanced cytotoxicity in laryngeal cancer cells through oxidative stress and apoptotic function on Hep2 cancer cells.

20-26

Authors: Vijayakumar Mayakrishnan, Selvi Veluswamy, Krishnakumari Shanmuga Sundaram, Priya Kannappan, Noorlidah Abdullah

Objective To elucidate free radical scavenging activity of ethanolic extract Lagenaria siceraria (L. siceraria) (Molina) fruit. Methods The free radical scavenging activity of the L. siceraria (Molina) fruit extract was assayed by using α,α-diphenyl-β-picrylhydrazyl (DPPH), 2,20-azinobis 3-ethyl benzothiazoline-6-sulfonate (ABTS), FRAP, reducing power, chelating ability and β-carotene bleaching assay. Results The IC50 values of DPPH and ABTS radical-scavenging activity was found to be 1.95 mg/mL and 19 mg/mL, respectively. In ferrous chelation assay, the percentage of inhibition was found to be 89.21%. The reducing power of ethanolic extract of L. siceraria (Molina) fruit was 0.068 at 1 mg/mL and increased to 0.192 at 5 mg/mL. The β-carotene linoleate bleaching assay was 46.7% at 5 mg/mL and antioxidant activity using FRAP at 0.305 for 1 mg/mL to 0.969 for 5 mg/mL. Conclusions The results indicate that L. siceraria (Molina) fruit could be an important sources of natural radical scavengers.

37-42

Authors: Jude C Ikewuchi

Objective To investigate the effects of an aqueous leaf extract of Acalypha wilkesiana (A. wilkesiana) on plasma chemistry and hematological indices of sub-chronic salt-loaded rats. Method The control group received a diet consisting 100% of the commercial feed, while the four test groups were received a diet consisting 8% salt and 92% commercial feed all through, except for the reference treatment group that had its salt-loading discontinued after six weeks. The extract was orally administered daily at 200 and 250 mg/kg body weight; while the test control, reference and control groups received appropriate volumes of water by the same route. Results The extract had no negative effects on markers of liver and kidney functions, produced hemoconcentration, significantly higher (P<0.05) plasma calcium and potassium levels, and significantly lower (P<0.05) plasma sodium and chloride levels in the test animals compared to test control. Conclusions This result supports the traditional use of A. wilkesiana in the management of hypertension and suggests that the extract may be a potassium sparing diuretic whose mechanism of antihypertensive action may be via alteration of plasma sodium and potassium balances or calcium mediated alteration in vascular muscle tone.

49-52

Authors: Hong-Tao Liu, Chun-Lei Lu

Objective To explore the influence of silencing Bcl-2 expression by small interfering RNA (siRNA) on Bcl-2 protein expression, cell apoptosis rate and radiosensitivity of gastric cancer BGC823 cells. Methods siRNA segment for Bcl-2 gene was designed and synthesized, then was induced into gastric cancer BGC 823 cells by liposome transfection. Bcl-2 protein expression was detected by Western Blotting. After X radiation, flow cytometry and clone forming assay were used to determine the effects of RNA interference on BGC823 cell apoptosis rate and radiosensitivity. Result After the transfection of Bcl-2 siRNA, the positive expression rate of Bcl-2 protein in BGC823 cells was (35.45±2.35)%. Compared with the control group and negative siRNA transfection group, the rate was significantly decreased (P<0.01). The apoptosis rate of BGC823-RNAi cell was (10.81±0.91)%, which was significantly higher than the control group and negative siRNA transfection group (P<0.01). After 48h X radiation, the apoptosis rate of BGC823-RNAi was (28.91±1.40)%, which was significantly higher than the control group and the group without radiation (P<0.01). During clone forming assay D0, Dq and SF2 values in Bcl-2 siRNA1 transfection group were all lower than those in the control group. The radiosensitivity ratio was 1.28 (the ratio of D0) and 1.60 (the ratio of Dq). Conclusions Specific siRNA of Bcl-2 gene can effectively inhibit the expression of Bcl-2 gene, enhance the radiosensitivity and apoptosis of gastric cancer BGC823 cells, having good clinical application perspective.

57-60

Authors: Ping Feng, Zhi-Lin Zhang, Zhi-Hua Zhang, Xiu-Long Zhang, Feng Xiang, Jian-Hua Tang, Bao-Li Xiang

Objective To evaluate the therapeutic effect of endostar (ED) combined with cisplatin(DDP) on model of C57BL/6 rats, and to further investigate the inhibiting mechanism of endostar from tumor angiogenesis. Methods Lewis lung cancer cells were inoculated in C57BL/6 mouse, then the mouse were randomized into control group (group A), ED (group B), DDP (group C) and ED/DDP (group D). They were treated according to the plan. And the expressions of VEGF and Sema3A were evaluated by immunhistochemisty. Results The weight of tumor increased in group A and B. It was decreased in group C and D. The tumor volume was increased in all the 4 groups. The VEGF expression of group D was obviously lower than the other group 3, but the Sema3A expressed of group D was significantly strengthener than the other group 3. The VEGF expression of group B and group D were obviously low especially in the 4th-8th days. Pearson correlated analysis showed that the expression VEGF and Sema3A were negatively correlated (r=-0.72, P<0.05). Conclusions ED combined with DDP could control the tumor growth effectively, and avoid weight loss. ED could reduce VEGF expression, and enhance Sema3A expression. Tumor vessel presents transient normalization. It is easy for DDP perfusion, and to kill tumor cells.

68-72

Authors: Feng Chen, Jian-Jun Li, Tao Liu, Guo-Qiang Wen, Wei Xiang

Objective To investigate clinical and neuroimaging features of enterovirus71 (EV71) related acute flaccid paralysis in patients with hand-foot-mouth disease. Methods Nine patients with acute flaccid paralysis met the criterion of EV71 induced hand-foot-mouth disease underwent spinal and brain MR imaging from May 2008 to Sep 2012. Results One extremity flaccid was found in four cases (3 with lower limb, 1 with upper limb), two limbs flaccid in three cases (2 with lower limbs, 1 with upper limbs), and four limbs flaccid in two cases. Spinal MRI studies showed lesion with high signal in T2-weighted images (T2WI) and low signal T1-weighted images (T1WI) in the spinal cord of all nine cases, and the lesions were mainly in bilateral and unilateral anterior horn of cervical spinal cord and spinal cord below thoracic 9 (T9) level. In addition, the midbrain, pons, and medulla, which were involved in 3 cases with brainstem encephalitis, demonstrated abnormal signal. Moreover, spinal cord contrast MRI studies showed mild enhancement in corresponding anterior horn of the involved side, and strong enhancement in its ventral root. Conclusions EV71 related acute flaccid paralysis in patients with hand-foot-mouth disease mainly affected the anterior horn regions and ventral root of cervical spinal cord and spinal cord below T9 level. MR imaging could efficiently show the characteristic pattern and extent of the lesions which correlated well with the clinical features.

76-78

Authors: Feng Liu, Fan Wang, Cong-Cong Wang, Nuo Li, Shu-Feng Li

Objective To explore the expression of IL-2 and IL-11 and its significance in patients with ankylosing spondylitis (AS). Methods A total of 48 active AS patients in our hospital and 40 normal control subjects were selected in our study. Bath ankylosing spondylitis disease activity index (BASDAI), Bath ankylosing spondylitis functional index (BASFI), Bath ankylosing spondylitis metrology index (BASMI), ESR and CRP expression levels were compared before treatment, 12 h after treatment and 24 h after treatment. IL-2 and IL-11 expression were also compared between these two groups. Results The BASDAI score, BASFI score and BASMI score of the AS patients before treatment significantly decreased compared with those 12 weeks and 24 weeks after treatment (P<0.05). ESR and CRP levels of the AS patients 12 weeks and 24 weeks after treatment significantly decreased compared with those before treatment (P<0.05). Difference was significant in serum IL-2 and IL-11 levels between 12 weeks and 24 weeks after treatment and before treatment (P<0.05). And no statistically significance was observed for serum IL-2 and IL-11 levels between normal control group and those of patients in AS group 24 weeks after treatment (P>0.05). Pearson's linear-correlation analysis showed that serum IL-2 level had a positive correlation with BASDAI, BASFI, BASMI, ESR and CRP (r=0.661,0.547,0.474,0.362,0.416, P<0.05) and serum IL-11 level had a negative correlation with BASDAI, BASFI, BASMI, ESR and CRP (r=-0.629, −0.412, −0.422, −0.387, −0.408, −0.315, P<0.05). Conclusions Serum levels of IL-2 in active AS patients significantly increase and will decrease after treatment. However, serum levels of IL-11 significantly decrease and will increase after treatment, which indicates that serum IL-2 has a positive correlation with the degree of AS and serum IL-11 has a negative correlation with the degree of AS, both of which are correlated closely with the onset of AS.

82-84

Authors: S Peter, N Malhotra, P Peter, R Sood

Dengue fever is a very common arthropod – borne infection in tropical countries. Neurological complications in dengue fever are relatively uncommon and among these, isolated cranial neuropathies have been reported only very rarely. We present an unusual neurological complication of Bell's palsy (lower motor neuron 7 th nerve palsy) associated with dengue infection. To the best of our knowledge, there have been very few documented cases of Flavivirus causing isolated Bell's palsy.

421-426

Authors: Pallara Janardhanan Wills, Velikkakathu Vasumathi Asha

To examine the downregulation of proinflammatory cytokines in a time dependant manner on carbon tetrachloride induced toxicity in experimental animals. Methods: CCl4 (150 毺L/100 g) was dissolved in corn oil (1:1 v/v %) and administered orally. Group I was treated as normal control and received corn oil on 8th day. Group II was toxic control and was given a single dose of CCl4 on 8th days. Group III was treated with Lygodium flexuosum (L. flexuosum) n-hexane extract (200 mg/kg) for 8 days and on 8th day a single dose of CCl4 was received. Group IV (negative control) received L. flexuosum n-hexane extract (200 mg/kg) alone for 8 days. Results: Treatment with n-hexane extract prior to the administration of CCl4 significantly prevented an increase in serum AST, ALT, LDH activity and lipid peroxidation and prevented the depletion of glutathione (GSH). Rats treated with L. flexuosum had reduced mRNA levels of TGF-毬1, TNF-毩 and IL-1毬 genes in liver of CCl4 intoxicated rats when compared to CCl4 control as evidenced by RT-PCR. Conclusions: The data suggest that L. flexuosum, a widely available fern, significantly reduces CCl4 induced acute hepatotoxicity by down-regulating the expression of pro-inflammatory cytokines in rats.

434-439

Authors: Chinnadurai Veeramani, Khalid S Al-Numair, Govindasamy Chandramohan, Mohammed A Alsaif, Kodukkur Viswanathan Pugalendi

To investigate the antihyperlipidemic effect of crude ethanolic extract of Melothria maderaspatana (M. maderaspatana) leaf (CEEM) on deoxycorticosterone acetate (DOCA)-salt hypertensive rats. Methods: A midscapular incision was made on each rat and the left kidney was excised after ligation of the renal artery. The surgical wound was closed using an absorbable suture. After one week recovery period, hypertension was induced by subcutaneous injection of DOCA-salt solution, twice a week, and the rats received a 1% sodium chloride solution as drinking water throughout the experimental period. CEEM or nifedipine was administered orally once a day for 6 weeks. Results: In DOCA-salt hypertensive rats, the level of plasma and tissues of total cholesterol (TC), triglycerides (TG), free fatty acids (FFA) and phospholipids (PL) significantly increased and administration of CEEM significantly reduced these parameters towards normality. Further, the levels of low density lipoprotein-cholesterol (LDL-C) and very low density lipoprotein-cholesterol (VLDL-C) significantly increased while high density lipoprotein-cholesterol (HDL-C) decreased in hypertensive rats and administration of CEEM brought these parameters to normality which proved their antihyperlipidemic action. Histopathology of liver, kidney and heart on DOCA-salt induced rats treated with CEEM showed reduced the damages towards normal histology. Conclusions: These findings provided evidence that CEEM was found to be protecting the liver, kidney and heart against DOCA-salt administration and the protective effect could attribute to its antihyperlipidemic activities.

448-454

Authors: Gino A Kurian, Jose Paddikkala

To evaluate pharmacological mimetic action of herbal extract Desmodium gangeticum (DG) roots on ischemia reperfusion injury. Methods: With the help of Langendroff perfusion technique, ischemic post condition (POC) mimetic action of DG methanol root extract was evaluated and compared by using standard drugs that acts as muscarinic receptor agonist and antagonist, namely acetylcholine (Ach) and atropine (Atr) respectively in an isolated rat heart. Results: The physiological parameters like left ventricular developed pressure, end diastolic pressure and working index of isolated rat heart showed significant recovery in DG root extract administrated rat heart, similar to the recovery by POC. Kymogram results showed muscarinic receptor agonist like action for DG methanol root extract, confirmed in rat heart by muscarnic receptor agonist (acetylcholine) and anatoginst (atropine). Administration of DG root extract prior to reperfusion showed better antioxidant status in myocardial tissue homogenate and mitochondrial, complemented by the levels of cardiac specific marker proteins in myocardial tissue and perfusate. Even though DG methanol root extract mimics its action similar to that of Ach, the myocardial protection mediated by the extract was superior to Ach, due to the presence of antioxidants in the crude extract. Conclusions: DG methanol root extract provides myocardial protection towards IRI by stimulating muscarinic receptors.

459-464

Authors: Shi Liu, Chao Yuan, Yun-Fu Cui, Bai-Xiang Li, Li-Jie Wu, Ying Liu

To investigate the precise species of tick vector and the Borrelia spirochete pathogen at the Heilongjiang Province international border with Russia. Methods: In this study, ticks were collected from 12 Heilongjiang border crossings (including grasslands, shrublands, forests, and plantantions) to determine the rate and species type of spirochete-infected ticks and the most prevalent spirochete genotypes. Results: The ticks represented three genera and four species of the Ixodidae family [Ixodes persulcatus, Dermacentor silvarum, Haemaphysalis concinna and Haemaphysalis japonica]. Ixodes persulcatus had the highest amount of Borrelia burgdorferi sensu lato infection of 25.6% and the most common species of Borrelia isolated from Ixodes persulcatus was Borrelia garinii, strain PD91. Conclusions: Our results suggest that Borrelia garinii PD91-infected Ixodes persulcatus may be the principal cause of Lyme disease in the border crossing areas of Heilongjiang Province.

469-471

Authors: Zhe Yan, Yang Liu, Hui Huang

To study the correlation of lipid ratios and individual lipid indexes of patients with type 2 diabetes with glycosylated hemoglobin (HbA1c). Methods: Samples were collected from 128 type 2 diabetic patients (aged 19-90 years; male 72, female 56). The sera were analyzed for HbA1c, total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). According to the HbA1c level, the patients were divided into three groups, group A (HbA1c <7%, n=31), group B (7%曑HbA1c曑10%, n=48), and group C (HbA1c >10%, n=49). The correlation of HbA1c with lipid ratios & individual lipid indexes were analyzed. Results: With the increased level of HbA1c, LDL-C had a significantly increasing trend (P<0.05); whereas TC went up with the increased HbA1c, without any significant differences between three groups. There was no significant correlation between HbA1c and TG or HDL-C. With the increased level of HbA1c, TC/HDL-C, LDL-C/HDL-C ratios were gradually increased, with significant differences among groups (P<0.05). The lipid ratios, especially LDL-C/HDL-C ratio was more susceptible to impaired lipid metabolism in T2DM patients than individual lipid. Conclusions: LDL-C/HDL-C ratio is helpful in assessing and reducing the risk of cardiovascular disease caused by impaired lipid metabolism in type 2 diabetic patients.

476-479

Authors: Jing Chen, Zi-Yi Guo, Guang Yang, Xiong Wang, Qing-Yu Tang, Yue-Qiong Cheng, Yi Guo, Shui-Xi Fu, Cai-Xiang Chen, Xiang-Jun Han

This article has been retracted: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The authors have plagiarized part of a paper that had already appeared in [European Journal of Radiology 81 (2012) 1851-1857. http://dx.doi.org/10.1016/j.ejrad.2009.09.017]. One of the conditions of submission of a paper for publication is that authors declare explicitly that their work is original and has not appeared in a publication elsewhere. Re-use of any data should be appropriately cited. As such this article represents a severe abuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.

485-497

Authors: Nisha Singh, Manish Kumar, Rakesh Kumar Singh

The control of Leishmania infection relies primarily on chemotherapy till date. Resistance to pentavalent antimonials, which have been the recommended drugs to treat cutaneous and visceral leishmaniasis, is now widespread in Indian subcontinents. New drug formulations like amphotericin B, its lipid formulations, and miltefosine have shown great efficacy to treat leishmaniasis but their high cost and therapeutic complications limit their usefulness. In addition, irregular and inappropriate uses of these second line drugs in endemic regions like state of Bihar, India threaten resistance development in the parasite. In context to the limited drug options and unavailability of either preventive or prophylactic candidates, there is a pressing need to develop true antileishmanial drugs to reduce the disease burden of this debilitating endemic disease. Notwithstanding significant progress of leishmanial research during last few decades, identification and characterization of novel drugs and drug targets are far from satisfactory. This review will initially describe current drug regimens and later will provide an overview on few important biochemical and enzymatic machineries that could be utilized as putative drug targets for generation of true antileishmanial drugs.

500-502

Authors: Jaydeep Sarkar, Aditi Chopra, Bhimrai Katageri, Hans Raj, Atul Goel

Leptospirosis, long thought to be a disease of sewage workers and found post-flooding is re-emerging as a potentially life threatening infectious disease in urban India during monsoon and immediately after due to poor sanitation and drainage. However, it is often ignored and under diagnosed due to a low index of suspicion. We present a fatal case of leptospirosis with acute liver failure, respiratory failure and jaundice and encephalopathy.