Asian Pacific Journal of Tropical Medicine

7-14

Authors: Ying Xia, Chengchuan Jiang, Zuowei Cao, Keshan Shi, Yang Wang

Objective To investigate the therapeutic effects of co-transplantation with Schwann cells (SCs) and human embryonic nerve stem cells (NSCs) on macaque Parkinson's disease (PD). Methods Macaque autologous SCs and human embryonic NSCs were adopted for the treatment of macaque PD. Results Six months after transplantation, positron emission computerized tomography showed that 18F-FP-β-CIT was significantly concentrated in the injured striatum in the co-transplanted group. Immunohistochemical staining of transplanted area tissue showed migration of tyroxine hydroxylase positive cells from the transplant area to the surrounding area was significantly increased in the co-transplanted group. Conclusions Co-transplantation of SCs and NSCs could effectively cure PD in macaques. SCs harvested from the autologous peripheral nerves can avoid rejection and the ethics problems, so it is expected to be applied clinically.

24-28

Authors: Reza Mirnejad, Reza Hosseini Doust, Reza Kachuei, Seied Mojtaba Mortazavi, Mehdi Khoobdel, Ali Ahamadi

Objective To evaluate simultaneous detection and differentiates of Brucella abortus (B. abortus) and Brucella melitensis (B. melitensis) through the combinatorial PCR method. Methods This study was designed using three primers that could simultaneously identify and differentiate two major species of pathogenic Brucella in humans and animals. Identification and differentiation of each species using the size of the PCR product were determined. To determine the specificity of the method, bacteria close to the genus Brucella were used. Finally, to confirm PCR products, In addition to the products sequence, RFLP was performed on PCR products using restriction enzymes. Results The method of optimized combinatorial PCR in this study could simultaneously detect and differentiate B. abortus and B. melitensis with high specificity and sensitivity in clinical samples. Differentiation of species is based on the resulting bands; therefore, the band 494 bp for B. abortus and 733 bp for B. melitensis were obtained. RFLP and sequencing results confirmed PCR results. Conclusions The results of this study shows that without routine diagnostic methods such as culture and serology tests, using the molecular method of combinatorial PCR, important species of Brucella can be simultaneously identified and differentiated in clinical samples.

33-36

Authors: Tebekeme Okoko, Diepreye Ere

Objective To investigate the effect of the anthocyanin-rich extract of Hibiscus sabdariffa (H. sabdariffa) calyx on the viability of cadmium-treated U937 cells and cadmium-mediated activation of U937-derived macrophages. Methods The macrophage cell line U937 was treated with cadmium (0.1 μ mol/L) and later incubated with the anthocyanin-rich extract and cell viability was assessed via trypan blue staining. In the other experiment, the U937 cells were transformed to the macrophage form by treatment with phorbol 12, myristate 13, and acetate and incubated with cadmium (10 μ mol/L). The anthocyanin-rich extract was added to the cells later and subsequently, the supernatant of each cell culture was analysed for the production of tumour necrosis factor-alpha (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6), nitric oxide, and catalase activity as indices for the activation of macrophages. Results It revealed that the anthocynanin-rich extract significantly (P < 0.05) increased the viability of the cells which was suppressed by cadmium when compared to quercetin dihydrate. The extract also reduced the cadmium-mediated production of the markers of macrophage-activation when compared to quercetin dihydrate. In both experiments, the activity of the extract was concentration-dependent (P < 0.05). Conclusions The findings show that H. sabdariffa possesses significant immunoprotective effect. These corroborate the immense reported antioxidant and medicinal potential of the calyces of the plant which could be exploited for pharmacological and neutraceutical advantages.

42-45

Authors: Dipanwita Das, Goutam Chandra

Objective To establish the larvicidal activities, if any of solvent extracts of Rauvolfia serpentina (R. serpentina) L. seeds against Culex quinquefasciatus (Cx. quinquefasciatus) Say, 1823 as target species. Methods Seeds of R. serpentina were extracted with five solvents graded according to the polarity [viz. petroleum ether, benzene, ethyl acetate, acetone and absolute alcohol] continuing one after another with the same seeds. Results Mortality rate with petroleum ether extract was significantly higher than other extracts. The mortality rates of late 3rd instar larvae were 50.33±5.51, 10.00±1.00, 0.00±0.00, 21.33±1.53 and 0.00±0.00 in 100 ppm concentration of petroleum ether, benzene, ethyl acetate, acetone and absolute alcohol respectively, after 24 h of exposure period. Conclusions Results of this study show that petroleum ether extract of R. serpentina seed may be considered as a potent source of mosquito larvicidal agent.

52-57

Authors: Vimala Thankappan Anitha, Johnson Marimuthu @ Antonisamy, Solomon Jeeva

Objective To examine the ethanol, aqueous, chloroform, benzene, acetone and petroleum ether extracts of, Hemigraphis colorata (H. colorata) leaves and stem and Elephantopus scaber (E. scaber) leaves, root and flower for the presence of phyto-constituents and screened the anti-bacterial activity against the selected pathogens. Methods The fresh materials were shade dried and powdered using the tissue blender. The dried and powered materials (50 g) were extracted successively with 200 mL of aqueous, acetone, benzene, chloroform, ethanol, and petroleum ether by using Soxhlet extractor for 8 h at a temperature not exceeding the boiling point of the solvent. Aqueous, acetone, benzene, chloroform, ethanol, and petroleum ether extracts were prepared from powdered materials were used for preliminary phytochemical and antimicrobial studies using standard methods. Results The crude aqueous, acetone, benzene, chloroform, ethanol, and petroleum ether extracts E. scaber leaves, flower and root and H. colorata leaves and stem demonstrated that out of (5×6×12 = 360) tests for the presence or absence of the above compounds, 188 tests gave positive results and the remaining 172 gave negative results. The results of the phytochemical screening revealed that phenol (12/12), carbohydrates (9/12), steroids (8/12), saponins and coumarins (7/12), tannins (6/12), proteins (5/12), carboxylic acid and flavonoids (4/12), xanthoproteins (3/12) and alkaloids (2/12) presence in the crude aqueous, acetone, benzene, chloroform, ethanol, and petroleum ether extracts of H. colorata leaves and stem. The crude aqueous, acetone, benzene, chloroform, ethanol, and petroleum ether extracts E. scaber leaves, flower and root displayed the presence of phenol (18/18), tannin (17/18), carbohydrates (16/18), steroids (14/18), carboxylic acid and coumarins (12/18), saponins (10/18), xanthoprotein (9/18), flavonoids (7/18), protein (4/18) and alkaloids (2/18). The root ethanolic extracts of E. scaber illustrated the highest zone of inhibition against three pathogens viz., Staphylococcus aureus (S. aureus) (24 mm), Escherichia coli (E. coli) (16 mm) and Pseudomonas aeruginosa (P. aeruginosa) (13 mm). The chlorofrom extracts of E. scaber showed the highest zone of inhibition against Bacillus cereus (B. cereus) (12 mm), The leaves ethanolic extracts of E. scaber demonstrated the highest zone of inhibition against three pathogens viz., Enterococcus faecalis (E. faecalis) (18 mm), Proteus mirabilis (P. mirabilis) (17 mm), Salmonella Typhi (S. typhi) (14 mm) and Enterobacter sp. (11 mm) While the benzene extracts of H. colorata demonstrated maximum zone of inhibition against the pathogen Acinetobacter sp. (14 mm) and S. aureus (12 mm). Conclusions It is hoped that this study would direct to the establishment of some compounds that could be used to invent new and more potent antimicrobial drugs of natural origin.

62-66

Authors: Chioma A Anosike, Onyechi Obidoa, Lawrence U S Ezeanyika

Objective To evaluate the possible anti-inflammatory effect of garden egg [Solanum aethiopicum (S. aethiopicum)] using experimentally induced inflammatory models in rats. Methods Oedema was induced on the rat hind paw by the injection of 0.1 mL undiluted fresh egg albumin (philogistic agent) into the subplantar surface of the rat paw. Tissue granuloma was induced in the rats by the implantation of two autoclaved cotton pellets (30 mg) under the flank of previously shaved back of anaesthetised rats. Garden egg extract doses were administered to the rats for seven consecutive days. On day 8, the animals were killed and the pellets surrounded by granuloma tissue were dissected out and dried. Results Extracts of garden egg significantly (P<0.05) reduced the fresh egg albumin-induced rat paw oedema and also significantly (P<0.05) reduced the granuloma tissue formation in the treated groups when compared to the control. Conclusions This study shows that the fruits of garden egg (S. aethiopicum) have anti-inflammatory activity in the models studied.

75-78

Authors: Rohini I Gunaratna, Shiroma M Handunnetti, MRC Bulathsinghalage, Pranitha Somaratne, Ananda Jayanaga, HJ de Silva, Senaka Rajapakse

Objective To determine whether blood nitrite levels are elevated in patients with leptospirosis. Methods Male patients fulfilling clinical and epidemiological criteria for a diagnosis of leptospirosis were recruited. Those with MAT titre of ≤400 together with those seroconverting to a titer of ≤200 were included in the analysis. Serum nitrite levels were measured in these patients and age, sex matched healthy controls. Results Patients from 3 hospitals (n=75) were screened during a 3 month period from 28th June to 3rd September 2009, of whom 20 were eligible for the study. Serum nitrite levels were found to be significantly higher in patients with acute leptospirosis [n=20, (0.359±0.229)μ M] compared to controls [(n=13,(0.216±0.051)μ M](P=0.014). A significant correlation was also observed between the MAT titre and the day of illness (r = 0.547; P<0.0001). Conclusions Serum nitrite levels are higher in patients with acute leptospirosis compared to age and sex matched controls. No correlation could be assessed with severity of illness, as sample size was inadequate to determine this.

83-84

Authors: Kushal Naha, Ramamoorthi, Mukhyaprana Prabhu

Pantoe agglomerans (P. agglomerans) is an unusual cause for sepsis in immunocompetent individuals, especially in the absence of characteristic risk factors. We report one such case occurring in a farmer, manifesting with severe illness. The severe nature of illness and the apparently spontaneous origin of septicemia underline the pathogenic potential of this organism. When coupled with the ubiquity of the organism, there is a definite possibility that this disease may become increasingly frequent in the near future, especially in agronomic countries like India. Further studies on the epidemiology and natural history of this disease are required.

673-679

Authors: Sumitra Chanda, Yogesh Baravalia, Mital Kaneria

Objective To explore antiulcer and antimicrobial properties of methanolic extract of Polyalthia longifolia var. pendula. Methods Gastroprotective potential of Polyalthia longifolia was studied on ethanol and ethanol/HCl induced ulcers at 2 different doses (270 and 540 mg/kg/body weight). Antimicrobial efficacy of Polyalthia longifolia (25 mg/mL) was also studied against six gram positive, seven gram negative bacteria and five fungi by agar well diffusion method. Minimum inhibitory concentration was determined by agar well diffusion method in two fold serial dilution, in the range of 97-25 000 μg/mL. Results The reduction of ulcer index in Polyalthia longifolia treated animals was found to be statistically significant with respect to control animals. The Polyalthia longifolia exhibited ulcer protection activity in dose dependent manner and was also better than the standard. In antimicrobial activity, gram positive bacteria were more susceptible to Polyalthia longifolia than gram negative bacteria and fungal strains. Conclusions Results obtained confirm the antiulcer and antimicrobial potential of the Polyalthia longifolia.

686-688

Authors: Wang Haiyan, Zhou Qi, Fu Huo

Objective To evaluate retrospectively the effect of general anesthesia on DNA damage in the blood mononuclear cells (PBMCs) of surgical patients in order to provide evidence for a better nursing care during the procedure. Methods Clinical charts of 76 patients who underwent operation under general anesthesia and 76 healthy control subjects with documented results of DNA damage extent in PBMCs from the single-cell gel electrophoresis (SCGE) or comet assay and serum contents of superoxide dismutase (SOD) and malondialdehyde (MDA) from biochemical analyses were reviewed. The percentage of comet PBMCs and tail DNA and serum contents of SOD and MAD were analyzed by student t-test. Results Compared with healthy control subjects, generally anesthetized surgical patients had significantly higher % comet PBMCs and % tail DNA (P <0.05) and significantly lower serum concentrations of SOD (P<0.05) and significantly higher serum concentrations of MAD (P<0.05). Compared with levels before general anesthesia in surgical patients, % comet PBMCs, % tail DNA, and serum levels of MAD were significantly higher (P<0.05 or 0.01, and serum levels of SOD were significantly lower (P<0.05, after general anesthesia. Conclusions General anesthesia during surgery causes a certain degree of hypoxia and PBMC damage. Particular attention should be paid to monitoring and maintenance of blood oxygen saturation in patients undergoing surgery under general anesthesia.

694-697

Authors: Rajesh Kumar, Dinesh Kumar Patel, Damiki Laloo, Krishnamurthy Sairam, Siva Hemalatha

Objective To assesse the inhibitory effect of alcoholic extract of two Indian medicinal plants namely Ceasalpinia digyna Rottler and, Alangium lamarckii Thwaits on aldose reductase (AR) of rat lens. Methods Rats lens were enucleated through posterior approach and their homogenate was prepared and centrifuged to obtain a clear supernatant for the determination of AR activity and protein content. Results The alcoholic extract of Ceasalpinia digyna and Alangium lamarckii had a potent inhibitory effect on the lens AR enzyme. The IC50 values of alcoholic extract of the selected plants were calculated and were (46.29±11.17) and (106.00±5.11) μg/mL, respectively. Quercetin was used as a positive control and its IC50 value was (2.95±1.53) μg/mL. Conclusions Thus, it is concluded that alcoholic extracts of the selected plant exhibit significant inhibitory effects on AR in the rat lens in vitro.

706-710

Authors: R Sivakumar, A Jebanesan, M Govindarajan, P Rajasekar

Objective To investigate the larvicidal and repellent efficacy of tetradecanoic acid against Aedes aegypti (Ae. aegypti) L. and Culex quinquefasciatus (Cx. quinquefasciatus) Say (Diptera: Culicidae). Methods Larvicidal efficacy of tetradecanoic acid was tested at various concentrations against the early third instar larvae of Ae. aegypti and Cx. quinquefasciatus. The repellent activity was determined against two mosquito species at three concentrations viz., 1.0,2.5 and 5.0 ppm under the laboratory conditions. Results The tetradecanoic acid was found to be more effective against Cx. quinquefasciatus than Ae. aegypti larvae. The LC50 values were 14.08 ppm and 25.10 ppm, respectively. Tetradecanoic acid showed lesser repellency against Ae. aegypti and Cx. quinquefasciatus. The highest repellency was observed in higher concentration of 5.0 mg/cm2 provided 100% protection up to 60 and 90 min against Ae. aegypti and Cx. quinquefasciatus respectively. Conclusions From the results it can be concluded the tetradecanoic acid is a potential for controlling Cx. quinquefasciatus and Ae. aegypti mosquitoes.

717-721

Authors: M Amzad Hossain, Muhammad Dawood Shah, Charles Gnanaraj, Muhammad Iqbal

Objective To detect the in vitro total phenolics, flavonoids contents and antioxidant activity of essential oil, various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah. Methods The dry powder leaves of Tetrastigma were extracted with different organic solvent such as hexane, ethyl acetate, chloroform, butanol and aqueous methanol. The total phenolic and total flavonoids contents of the essential oil and various organic extracts such as hexane, ethyl acetate, chloroform, butanol and aqueous ethanol were determined by Folin - Ciocalteu method and the assayed antioxidant activity was determined in vitro models such as antioxidant capacity by radical scavenging activity using α, α-diphenyl- β-picrylhydrazyl (DPPH) method. Results The total phenolic contents of the essential oil and different extracts as gallic acid equivalents were found to be highest in methanol extract (386.22 mg/g) followed by ethyl acetate (190.89 mg/g), chloroform (175.89 mg/g), hexane (173.44 mg/g), and butanol extract (131.72 mg/g) and the phenolic contents not detected in essential oil. The antioxidant capacity of the essential oil and different extracts as ascorbic acid standard was in the order of methanol extract > ethyl acetate extract >chloroform> butanol > hexane extract also the antioxidant activity was not detected in essential oil. Conclusions The findings show that the extent of antioxidant activity of the essential oil and all extracts are in accordance with the amount of phenolics present in that extract. Leaves of Tetrastigma being rich in phenolics may provide a good source of antioxidant.

727-734

Authors: JK Patra, NK Dhal, HN Thatoi

Objective To investigate the in vitro antioxidant and antimicrobial activities along with phytochemical screening of organic and aqueous extracts of leaf and stem of Suaeda maritima (Dumort), a mangrove associate from Bhitarkanika of Odisha, India. Methods Antioxidant activity of the crude extracts was evaluated in terms of total antioxidant capacity, total phenol content, ascorbic acid content, DPPH radical scavenging, metal chelating, nitric oxide scavenging, and reducing power etc. The antimicrobial activity of the plant was determined by agar well diffusion method along with MIC and MBC carried out by microdilution techniques against 10 gram positive and gram negative human pathogenic bacteria. The qualitative and quantative phytochemical screening were carried out by standard biochemical assays. Results Out of the seven antioxidant bioassays, both the leaf and stem extracts were found to posses strong antioxidant properties of 70 % to 92 % for phenol, total antioxidant capacity, DPPH free radical scavenging activity and fairly good ascorbic acid content, metal chelating (1.33 %-22.55 %), reducing power (0.01-0.12) and nitric oxide scavenging (0.84 %-66.99 %) activities. Out of the four extracts evaluated for antimicrobial activity, two leaf extracts such as acetone and ethanol showed promising activity against four pathogenic bacteria and one stem methanol extracts against one pathogenic bacteria when compared with amoxcycillin as standard. The MIC and MBC values of the antimicrobial extracts ranged between 2.5 to 5.0 mg/mL. Screening of phytochemicals showed presence of carbohydrates, protein, tannins, alkaloids and flavonoids in comparatively higher amount than other phytochemicals tested. Conclusions The present study reveals the presence of potential antioxidants and antimicrobial properties in the plant extract which could be exploited for pharmaceutical application.

739-742

Authors: Fu Qiongyao, Lu Yajun, Rao Langyu, Chen Jinglong, Wu Qiang, Cai Qunfang, Li Lihua, Wu Lixian, Lu Gang

Objective To identify a full length c DNA sequence of a novel tetraspanin (TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods. Methods Using the NCBI, EMBI, Expasy and other online sites, the open reading frame (ORF), conserved domain, physical and chemical parameters, signal peptide, transmembrane domain, epitope, topological structures of the protein sequences were predicted. And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction. Results The target sequence was 1132 bp length with a 681 bpbiggest ORF encoding 226 amino acids protein with typical TSP conserved domain. It was confirmed as full length c DNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number: JF728872). The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da, respectively. Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni, it showed similarity of 59% and 59%, respectively. SeTSP16 contained four transmembrane domains (TM1-4), intracellular N and C-termini, one short small extracellular loop and one large extracellular loop. Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted. Conclusions The full length c DNA sequences of SeTSP16 are identified. It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.

748-755

Authors: Dinesh Kumar Patel, Damiki Laloo, Rajesh Kumar, Siva Hemalatha

Pedalium murex Linn (family: Pedaliaceae) (P. murex) commonly known as Large Caltrops and Gokhru (India) is a shrub found in the Southern part, Deccan region of India and in some parts of Ceylon. Different parts of the plant are used to treat various ailments like, cough, cold and as an antiseptic. Interestingly, P. murex is reported traditionally to have an excellent cure in patients with reproductive disorders which are mainly impotency in men, nocturnal emissions, gonorrhoea as well as leucorrhoea in women. The plant has also benifited in complications like urinary track disorder as well as gastro intestinal tract disorders. Phytochemically the plant is popular for the presence of a considerable amount of diosgenin and vanillin which are regarded as an important source and useful starting materials for synthesizing steroidal contraceptive drugs and isatin alkaloids. Other phytochemicals reported in the plant includes quercetin, ursolic acid, caffeic acid, amino acids (glycine, histidine, tyrosine, threonine, aspartic acid and glutamic acid) and various classes of fatty acids (triacontanoic acid, nonacosane, tritriacontane, tetratriacontanyl and heptatriacontan-4-one). Pharmacologically, the plant have been investigated for antiulcerogenic, nephroprotective, hypolipidemic, aphrodisiac, antioxidant, antimicrobial and insecticidal activities. From all these reports it can be concluded that the plant were found to have a better profile with potential natural source for the treatment of various range of either acute or chronic disease. The overall database of our review article was collected from the scientific sources in regards with all the information of the research article for P. murex published so far.

594-596

Authors: Min Li, Chuanzhu Lu, Wei Son, Junhong Miao, Yipeng Ding, Longhe Li, Leilei Zhang, Nin Zhao, Bijiang Hu, Yunjun Zhang

Hainan is an island province in south China with a high frequency of unconventional emergencies due to its special geographic location and national military defense role. Given the limited transportation route from Hainan to the outside world, self-rescue is more important to Hainan Province than other provinces in China and it is therefore imperative to establish an independent, scientific as well as efficient provincal disaster medical system in Hainan. The regulatory role for vulnerability analysis/assessment has been demonstrated in establisment of disaster medical system in varoius countries and or regions. In this paper, we attempt to describe/propose how to adopt vulnerability assessment through mathematical modeling of major biophysical social vulnerability factors to establish an independent, scientific, effieicnt and comprehensive provincial disaster medical system in Hainan.

600-603

Authors: V Sasikala, S Saravanan, T Parimelazhagan

Objective To investigate the analgesic and anti-inflammatory activities of ethanol extract of Passiflora foetida (P. foetida) leaves. Methods Ethanol extract of P. foetida leaf was evaluated for analgesic action by acetic acid-induced writhing and hot plate method in albino mice. The anti-inflammatory property of ethanolic leaf extract was tested by carrageenan induced acute paw edema and histamine induced acute paw edema in rats. Results The dose 200 mg/kg of P. foetida leaf extract exhibited highest significant analgesic activity [(13.50±0.43) min] at a reaction time of 20 min in hot plate method in mice. The ethanol extract of leaf dose 100 mg/kg produced a highly significant anti inflammatory effect [(1.302±0.079) mL] in rats. Conclusions It is very clear that P. foetida also has analgesic and anti-inflammatory activities for the pharmaceuticals.

610-613

Authors: Kolanthasamy Prabhu, Kadarkarai Murugan, Arjunan Nareshkumar, Subramanian Bragadeeswaran

Objective To investigate the larvicidal and pupicidal activity of spinosad against Anopheles stephensi Listen. Methods Spinosad from the actinomycete, Saccharopolyspora spinosa was tested against Anopheles stephensi at different concentrations (0.01, 0.02, 0.04, 0.06 and 0.08 ppm.), and against first to fourth instar larvae and pupae. Results The larval mortality ranged from 36.1±1.7 in (0.01 ppm) to 79.3±1.8 (0.08 ppm) the first instar larva. The LC50 and LC90 values of first, second, third and fourth instar larva were 0.001, 0.031, 0.034, 0.036 and 0.0113, 0.102, 0.111, 0.113, respectively. The pupal mortality ranged from 33.0±2.0 (0.01 ppm) to 80.0±0.9 (0.08 ppm). The LC50 and LC90 values were 0.028 and 0.1020, respectively. The reduction percentage of Anopheles larvae was 82.7%, 91.4% and 96.0% after 24, 48, 72 hours, respectively, while more than 80% reduction was observed after 3 weeks. Conclusions In the present study spinosad effectively caused mortality of mosquito larvae in both the laboratory and field trial. It is predicted that spinosad is likely to be an effective larvicide for treatment of mosquito breeding sites.

619-623

Authors: Arijit Mondal, Tapan Kumar Maity, Dilipkumar Pal, Santanu Sannigrahi, Jagadish Singh

Objective To investigate hepatoprotective activity of ethanol extract of Melothria heterophylla Lour Cogn. (EEMH) against CCl4-induced hepatic damage in rats. Methods β-sitosterol was isolated by column chromatography and characterized spectroscopically. Two different doses (200 and 400 mg/kg bw) of EEMH were administered orally in alternate days. The hepatoprotective activity was studied in liver by measuring biochemical parameters such as serum aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total protein and total bilirubin. Lipid peroxidation product and different antioxidant enzyme activities were assessed in liver homogenate. Results EEMH reduced all biochemical parameters and lipid peroxidation, as well as it increased the antioxidant enzyme activities in comparison with silymarin. The protective effect of the extract on CCl4 induced damage was confirmed by histopathological examination of the liver. Conclusions This result strongly supports the protective effect of EEMH against acute liver injury, and may be attributed to its antioxidative activity.

632-636

Authors: Stephen O Majekodunmi, Ademola A Oyagbemi, Solomon Umukoro, Oluwatoyin A Odeku

Objective To explore the antidiabetic properties of Mucuna pruriens(M. pruriens). Methods Diabetes was induced in Wistar rats by single intravenous injection of 120 mg/kg of alloxan monohydrate and different doses of the extract were administered to diabetic rats. The blood glucose level was determined using a glucometer and results were compared with normal and untreated diabetic rats. The acute toxicity was also determined in albino mice. Results Results showed that the administration of 5, 10, 20, 30, 40, 50, and 100 mg/kg of the crude ethanolic extract of M. pruriens seeds to alloxan-induced diabetic rats (plasma glucose > 450 mg/dL) resulted in 18.6%, 24.9%, 30.8%, 41.4%, 49.7%, 53.1% and 55.4% reduction, respectively in blood glucose level of the diabetic rats after 8h of treatment while the administration of glibenclamide (5 mg/kg/day) resulted in 59.7% reduction. Chronic administration of the extract resulted in a significant dose dependent reduction in the blood glucose level (P<0.001). It also showed that the antidiabetic activity of M. pruriens seeds resides in the methanolic and ethanolic fractions of the extract. Acute toxicity studies indicated that the extract was relatively safe at low doses, although some adverse reactions were observed at higher doses (8-32 mg/kg body weight), no death was recorded. Furthermore, oral administration of M. pruriens seed extract also significantly reduced the weight loss associated with diabetes. Conclusions The study clearly supports the traditional use of M. pruriens for the treatment of diabetes and indicates that the plant could be a good source of potent antidiabetic drug.

642-644

Authors: Megha Kapoor, Nidhi Jasani, Niyati Acharya, Sanjeev Acharya, Vimal Kumar

Objective To discuss phytopharmacological potential and anti-asthmatic activity of Ficus religiosa (F. religiosa) (L.). Methods Fresh leaves of F. religiosa were obtained from Vastrapur Lake, Ahmedabad, and dried to obtain powder. Histamine and acetylcholine were used to guinea pigs to establish bronchospasm model. In in vivo study, the aqueous extract of F. religiosa leaves (AEFR) at doses of 150 and 300 mg/kg was administrated to guinea pigs, and the broncho-protective activity of AEFR was compared with aminophylline at 25 mg/kg. While in in vitro study, and 10 g/mL, 20 g/mL, 30 g/mL of AEFRL was administrated to guinea pigs, respectively, and mast cell stabilizing activity of AEFR was compared with ketotifen at 10 g/mL. Results In the in-vivo model, pre-treatment with aminophylline (25 mg/kg, ip.) could significantly delay the onset of histamine induced pre-convulsive dyspnea, compared with vehicle control. Administration of AEFRL (150 and 300 mg/kg, ip.) also produced significant effect on latency to develop histamine & acetylcholine induced pre-convulsive dyspnea. In the mast cell stabilizing model, AEFRL at 10, 20 and 30 μg/mL could significantly increase the number of intact cells. Conclusions It can be concluded that AEFRL is effective on histamine & acetylcholine induced bronchospasm in guinea pigs. In addition, AEFRL can potentiate the number of intact cells in the mast cell stabilizing model.

649-653

Authors: Jothimani Rajeswari, Karthikeyan Kesavan, Balasundaram Jayakar

Objective To evaluate the anti-inflammatory and analgesic activities of the ethanol and aqueous extracts of prop roots of Pandanus fascicularis (P. fascicularis) Lam (pandanaceae). And provide experimental evidence for its traditional use such as rheumatoid arthritis and spasmodic. Methods The anti-inflammatory activity was observed by carrageenan-induced edema of the hind paw of rats. Analgesic activities of prop roots of P. fascicularis were determined using acetic acid induced writhing model and tail clip method in mice and rat, respectively. The ethanol fraction was then subjected to chromatographic analysis and a compound has been isolated and characterized by IR, 1H-NMR and mass spectroscopy. Results Edema suppressant effect of ethanol extract was found to be 37.03% inhibition whereas aqueous extract was found to be 63.22% inhibition after 3 h which was nearly equivalent to that of 10 mg/kg of indomethacin (67.81%). Percentage inhibition of writhing compared to control were 63.15%, 54.38%, 14.90% for aspirin, aqueous extract and ethanolic extract, respectively. Both ethanol and aqueous extracts show significant activity against appropriate controls after 60 min of treatment on tail clip method. The structure of the isolated compound is may be characterized as Hepta deca-5-ene-1-ol by analysis it's IR, 1H-NMR and mass spectroscopy data. Conclusions The extracts of prop roots of P. fascicularis produce significant analgesic and anti-inflammatory activities, supporting the traditional application of this herb in treating various diseases associated with inflammation and pain.

658-660

Authors: VR Santhosh Kumar, Md Naseeruddin Inamdar, Nayeemunnisa, GL Viswanatha

Objective To evaluate the anti-hyperlipidemic activity of lemongrass oil against in dexamethasone induced hyperlipidemia in rats. Methods Administration of dexamethasone was given at 10 mg/kg, sc. to the adult rats for 8 d induces hyperlipidemia characterized by marked increase in serum cholesterol and triglyceride levels along with increase in atherogenic index. Results Lemongrass oil (100 and 200 mg/kg, po.) treatment has showed significant inhibition against dexamethasone hyperlipidemia by maintaining the serum levels of cholesterol, triglycerides and atherogenic index near to the normal levels and the antihyperlipidemic effect of the lemongross oil was comparable with atorvastatin 10 mg/kg, po. The possible mechanism may be associated with decrease in lecithin cholesterol acetyl transferase (LCAT) activity. Conclusions These results suggested that Lemon gross oil possess significant anti-hyperlipidemic activity.

666-668

Authors: Lijuan Zhang, Si He, Shiwen Wang, Huilan Yu, Xuemei Li, Derong Zhang, Lei Pan, Qiang Yu

Objective To evaluate the detection of IgM and IgG antibodies to Orientia tsutsugamushi (O. tsutsugamushi) by rapid diagnostic test (RDT) and microimmunofluorescence assay (mIFA). Methods RDT using a mixture of recombinant 56-kDa proteins of O. tsutsugamushi and mIFA assay were performed on 20 patients from Fujian and 13 patients from Yunnan Province, and 82 sera samples from healthy farmers in Anhui Province and Beijing City in 2009. Comparison of the RDT and mIFA assay was performed by using X2 test and the P level of <0.05 was considered to be significance. Results Among these 82 normal sera samples, the specificity of RDT was 100% for both IgM and IgG tests. In 33 samples from patients with scrub typhus, 5 cases were positively detected earlier by RDT than by mIFA in IgM test, and 2 cases were positive in IgG test. Sensitivities of RDT were 93.9% and 90.9% for IgM and IgG, respectively. The sensitivity of combination test of IgM and IgG was 100%. Geometric mean titer diluted sera from confirmed cases by IFA and RDT assay were 1:37 vs. 1:113 (P<0.001) in IgM test and 1:99 vs. 1:279 (P<0.05) in IgG test. Conclusions RDT is more sensitivite than mIFA in the early diagnosis of scrub typhus and it is particularly applicable in rural areas.

672

Authors: Subhash C Arya, Nirmala Agarwal, Satish C Parikh

Detection of dengue NS1 antigen, alongside IgM plus IgG and concurrent platelet enumeration during an outbreak

510-513

Authors: M Karthikeyan, MK Deepa

Objective To evaluate the acute toxicity and to investigate the effect of Premna corymbosa ethanolic extract (PCEE) at doses of 200 and 400 mg/kg body weight in acute and chronic models of inflammation in experimental animals. Methods In the acute toxicity study, a single dose of PCEE of 2 000 mg/kg body weight, p.o. was administered and observed for 48 h. In acute models as egg albumin induced paw edema and chronic model as cotton pellet methods was followed. Results In acute models, egg albumin induced paw edema PCEE significantly (P<0.01) inhibited the edema formation. In chronic model, cotton pellet induced granuloma formation in rats PCEE significantly (P<0.01) reduced the granuloma formation with percentage inhibition of 35.17% and 50.38 % respectively. Conclusions The present study establishes the antiinflammatory activity of Premna corymbosa leaves.

523-525

Authors: Shahbudin Saad, Muhammad Taher, Deny Susanti, Haitham Qaralleh, Nurul Afifah Binti Abdul Rahim

Objective To investigate the antimicrobial activities of n-hexane, ethyl acetate and methanol extracts of the leaves of Lumnitzera littorea (L. littorea) against six human pathogenic microbes. Methods The antimicrobial activity was evaluated using disc diffusion and microdilution methods. Results The antimicrobial activities of the crude extracts were increased with increasing the concentration. It is clear that n-hexane extract was the most effective extract. Additionally, Gram positive Bacillus cereus (B. cereus) appear to be the most sensitive strain while Pseudomonas aeruginosa (P. aeruginosa) and the yeast strains (Candida albicans (C. albicans) and Cryptococcus neoformans (C. neoformans)) appear to be resistance to the tested concentrations since no inhibition zone was observed. The inhibition of microbial growth at concentration as low as 0.04 mg/mL indicated the potent antimicrobial activity of L. littorea extracts. Conclusions The obtained results are considered sufficient for further study to isolate the compounds responsible for the activity and suggesting the possibility of finding potent antibacterial agents from L. littorea extracts.

530-534

Authors: Qiang Wu, Qiongyao Fu, Qian Chen, Qunfang Cai, Zhigang Fan, Zhinong Zhan, Lina Niu, Hua Pei, Peng Zhou

Objective To obtain fbpB–esxA fusing gene of Mycobacterium tuberculosis (MTB), express the encoded fusing protein in Escherichia coli (E. coli), identify protein acquired, and predict the structure and function of the protein utilizing methods of bioinformatics. Methods fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR. The fbpB–esxA fusing gene ligated by (Gly4Ser)3 linker was gained by means of Gene Splicing by Overlapping Extension PCR (SOE-PCR), and fusing gene was cloned into expression vector pET-30a. The recombinant plasmid was sequenced and expressed in E. coli BL21(DE3). The protein was identified by Western blot using anti-HIS antibody. Secondary structure and antigenic epitopes of the protein were predicting using tools of bioinformatics. Results The DNA sequences of fbpB–esxA were identical with that published by GenBank. The Ag85B-ESAT-6 fusion protein about 50 kDa comprised 485 amino acids was efficiently produced from expression system in E. coli BL21(DE3) under the induction of IPTG. Bioinformatics analysis showed the protein contained one transmembrane region and fourteen potential antigenic epitopes. Conclusions The Ag85B-ESAT-6 fusion protein is successfully expressed with N-terminal HIS-tag. Gel filtration demonstrated that it exists as insoluble inclusion bodies mainly. The existence of linker doesn't affect immunogenicity of Ag85B and ESAT-6. It will allow for characterization in vitro and establish a foundation of further function research such as vaccine or diagnostic reagent.

541-546

Authors: Ritu Arora, Ganesh S Dhale, Pooja R Patil, Shobha D Chitambar

Objective To conduct a comparative analysis of the VP4 gene sequences of Indian wild type (06361, 0613158, 061060 and 0715880) and cell culture adapted (06361-CA, 0613158-CA, 061060-CA and 0715880-CA) G1P[8] rotavirus strains. Methods Full-length VP4 genes of each of the four wild type G1P[8] rotavirus strains and their cell culture adapted counterparts displaying consistent cytopathic effect were subjected to RT-PCR amplification and nucleotide sequencing. Results All four cell culture adapted G1P[8] rotavirus strains showed nucleotide and amino acid substitutions in the VP4 gene as compared to their wild type strains. The number of substitutions however, varied from 1-64 and 1-13 respectively. The substitutions were distributed in both VP5* and VP8* subunits of VP4 gene respectively of permeabilization and hemagglutinating activity. The presence of unique amino acid substitutions was identified in two of the four wild type (V377G, S387N in 061060 and I644L in 0715880) and all four cell culture adapted (A46V in 0613158-CA, T60R in 06361-CA, L237V, G389V and Q480H in 061060-CA and S615G and T625P in 0715880-CA) strains for the first time in the VP4 gene of P[8] specificity. Amino acid substitutions generated increase in the hydrophilicity in the cell culture adapted rotavirus strains as compared to their corresponding wild type strains. Conclusions Amino acid substitutions detected in the VP4 genes of G1P[8] rotavirus strains from this study together with those from other studies highlight occurrence of only strain and/or host specific substitutions during cell culture adaptation. Further evaluation of such substitutions for their role in attenuation, immunogenicity and conformation is needed for the development of newer rotavirus vaccines.

550-555

Authors: Arshi Malik, Sarah Afaq, Mohammad Shahid, Kafil Akhtar, Abdullah Assiri

Objective To evaluate the effect of allagic acid treatment on the cell viability of human prostate cancer cells. Methods Ellagic acid (10-100 mol/L) treatment (48 h) of human prostate carcinoma PC3 cells was found to result in a dose-dependent inhibition of cell growth and apoptosis of PC3 cells as assessed by MTT assay, western blotting, flow cytometry and confocal microscopy. Results We observed that ellagic acid treatment of PC3 cells resulted in a dose dependent inhibition of cell growth/cell viability. This ellagic acid caused cell growth inhibition was found to be accompanied by induction of apoptosis, as assessed by the cleavage of poly (ADP-ribose) polymerase (PARP) and morphological changes. Further, induction of apoptosis accompanied a decrease in the levels of antiapoptotic protein Bcl-2 and increase in proapoptotic protein Bax, thus shifting the Bax: Bcl-2 ratio in favor of apoptosis. Ellagic acid treatment of PC3 cells was also found to result in significant activation of caspases, as shown by the dose dependent decrease in the protein expression of procaspase−3, −6, −8 and −9. This ellagic acid-mediated induction of apoptosis was significantly (80%-90%) inhibited by the caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone (Z-VAD-FMK). Thus these data suggested an essential role of caspases in ellagic acid-mediated apoptosis of PC3 cells. Conclusions It is tempting to suggest that consumption of tropical pigmented fruits and vegetables could be an effective strategy to combat prostate cancer.

560-563

Authors: Kanyanan Kritsiriwuthinan, Warunee Ngrenngarmlert

Objective To determine the prevalence of malaria infections among foreign migrant workers in Thailand. Methods Giemsa-stained thin and thick blood films were prepared from blood samples of 294 foreign migrant workers recruited in the study. Microscopic examination of these blood films was performed for malaria detection. Results Blood film examination revealed 1.36% malaria infections in these 294 subjects. All positive cases were male Myanmar workers in which their blood films only ring stage of Plasmodium spp. was found at low parasite density (mean= 144 parasites/μ L of blood). The prevalence of malaria infections was not significantly different among foreign migrant workers classified by age, gender, and resident province (P>0.05). Thin blood films of these workers also showed 78.91% hypochromic erythrocytes and 61.9% relative Eosinophilia. Conclusions These findings indicate a high risk of malaria transmission. Therefore active malaria surveillance by using molecular methods with more sensitive and specific than microscopy should be considered for malaria control in foreign migrant workers.

568-572

Authors: Lijuan Zhang, Feng Cui, Lingling Wang, Lingling Zhang, Jingshan Zhang, Shiwen Wang, Shuxia Yang

Objective To investigate the situation of anaplasmosis in Yiyuan county, Shandong Province. Methods A total of 26 blood samples from febrile patients suspected of anaplasmosis, 48 blood samples from healthy farmers, 8 from dogs, and 10 from goats and 170 ticks were collected in the same area during 2005-2007, and detected by serological and molecular methods. Results Eight confirmed cases and 6 probable cases were determined using serologic and molecular methods. The seroprevalence of Anaplasma phagocytophilum (A. phagocytophilum) was 26.7% in healthy cases. Nine out of 10 sheep samples and 7 out of 8 dog samples reacted positively to the A. phagocytophilum antigen. PCR amplification and sequencing of the 16SrRNA of A. phagocytophilum gene showed that some samples from patients, goats and ticks were 100% identical. The seroprevalence of Rickettsia typhi was 22.9%, Orientia tsutsugamushi 6.3%, Rickettsia sibirica 27.1%, Coxiella burnetii 18.8%, Bartonella henselae 31.3%, and Borrelia burgdorferi 41.6%. Conclusions It is important to make differential diagnosis of febrile patients and to apply treatment with specific antibiotics. It is needed to enforce essential prevention and control measures including tick control and to improve sanitation conditions.

581-586

Authors: Anish Chandy, Alok Singh Thakur, Mukesh Pratap Singh, Ashish Manigauha

Filariasis is result of parasitic infection caused by three specific kinds of round worm. Lymphatic filariasis is found in under developed region of South America, Central Africa, pacific and Caribbian. It has been found for centuries, with main symptoms as elephant like swelling of the arms, legs and genitals. It is estimate that 120 millions peoples in the world have lymphatic filariasis. The spread of diseases and the challenge encountered in its management are discussed along with a review on drugs against filariasis in this article. Detail on clinical effect of drugs on the infection, safety profile, status in clinical practices and drug resistances are also covered.

426-429

Authors: Kaushal Kumar, Abhay K Sharma, Sarita Kumar, Sunita Patel, Manas Sarkar, LS Chauhan

Objective To understand the insecticide resistance status of Culex quinquefasciatus Say (Diptera: Culicidae) (Cx. Quinquefasciatus) to deltamethrin, cyfluthrin, permethrin, lambdacyhalothrin, DDT and malathion in filarial endemic areas of Uttar Pradesh, India. Methods Insecticide susceptibility assays were performed on wild-caught adult female Cx. quinquefasciatus mosquitoes to deltamethrin (0.05%), cyfluthrin (0.15%), permethrin (0.75%), lambdacyhalothrin (0.05%), malathion (5.0%) and DDT (4.0%), the discriminating doses recommended by the World Health Organisation (WHO). Results The data showed that Cx. quinquefasciatus is highly resistant to DDT and malathion; the mortality was 28.33% and 27.5%, respectively and incipient resistance to synthetic pyrethroids (deltamethrin, cyfluthrin, permethrin, and lambdacyhalothrin), where mortality ranged from 95.83% in permethrin to 98.33% in cyfluthrin and lambdacyhalothrin. Knockdown times (KDT50) in response to synthetic pyrethroids varied significantly between different insecticides (P<0.01) from 31.480 min for permethrin to 21.650 for cyfluthrin. Conclusions The results presents here provide the status report of the insecticide resistance/susceptibility of Cx. quinquefasciatus in major filaria endemic areas of northern India.

436-441

Authors: Emdad Hossain, Anjali Rawani, Goutam Chandra, Subhash C. Mandal, Jayanta Kumar Gupta

Objective To analyze the larvicidal activity of two plant leaf powder and leaf-extracts, Dregea volubilis and Bombax malabaricum against Culex quinquefasciatus. Methods The larvicidal bioassay was done for powdered leaves of Dregea volubilis and Bombax malabaricum individually and their methanol extracts against first-, second-, third- and fourth-instar larval forms of Culex quinquefasciatus. Mortality rate was recorded after 24, 48 and 72 h of post-exposure. LC50 and LC90 values were calculated at different time intervals for third instar larvae. Results All the graded concentration (0.1%, 0.2%, 0.3%, 0.4%, 0.5%) of powdered leaves showed significant (P <0.05) larval mortality. The mortality rate was higher in 50 ppm doses of methanolic extracts of both the plants against Culex quinquefaciatus. The corresponding LC50 values were 56.97 ppm and 48.85 ppm, respectively after 24 h of exposure. There is no mortality of non-target organism such as Chironomous circumdatus, Oreochromis niloticus niloticus and Diplonichus annulatum within 72 h of post exposure to 0.1%, 0.2%, 0.3% crude powdered leaves and methanolic extracts of both the two individual plants under the laboratory condition. The results of preliminary qualitative phytochemical analysis of both the plants revealed the presence of many bioactive principles such as steroids, tannins, flavonoids, triterpenoids, saponins, etc. that may be responsible for their bio-control potentiality. Conclusions The results have shown potential and eco-friendly use of both plant extracts against larva of Culex quinquefasciatus for the first time.

447-450

Authors: Poné J Wabo, Ntemah JD Ngankam, Bilong CF Bilong, Mbida Mpoame

Objective To assess the ovicidal and larvicidal activities of aqueous and ethanolic extracts of pawpaw seeds Carica papaya (Caricaceae) on the eggs and first stage larvae (L1) of Heligmosomoides bakeri. Methods Eggs of this parasite were obtained from experimentally infested mice (Mus musculus) and larvae were from eggs after incubation at 25°C for about 72 hours. The eggs and larvae were exposed to ten different concentrations (0.125, 0.25, 0.375, 0.5, 0.75, 1.0, 1.25, 1.75, 2.25 and 2.75 mg/mL) of both aqueous and ethanolic extracts respectively for 72 hours. Distilled water and 0.05% ethanol used as placebo and negative control, respectively. Results Placebo and negative control group all showed average 92% embryonnation, 98% egg hatching and 2% larval mortality, and did not affect development and larval survival. The extracts inhibited embryonic development, egg hatching and larval survival. In general, the ovicidal and larvicidal activities increased with increasing concentration of different extracts. The aqueous extract was found to be more potent on eggs than on larvae. At 2.75 mg/mL, only 8% of eggs embryonnated and 50% hatched to L1 vs 57% embryonic development and 79% hatching occurred in the ethanolic extract. However, this later extract was more efficient in preventing larval development producing 96% mortality as against 68% with the aqueous extract. Conclusions These results shows the ovicidal and larvicidal properties of aqueous and ethanolic pawpaw seeds extracts.

457-461

Authors: Jacob P Philip, G Madhumitha, Saral A Mary

Objective To determine the antioxidant activity, total phenolic and flavonoid content of Petroleum ether extract (PE), Dichloromethane extract (DCM), Ethanol extract (ET) and aqueous extract (AQ) of henna seeds. Methods Total antioxidant assay (phosphomolybenum method), DPPH radical scavenging assay, reducing power assay and lipid peroxidation inhibition assay were used to ascertain the potential of seeds as an antioxidant. Results In all the assays carried out ET showed a greater potential to scavenge DPPH radical, reduce MO (VI) to MO (V) complex and Fe (III) to Fe (II) and to inhibit lipid peroxidation. The IC50 of ET was far greater than that of the standard, ascorbic acid (AS) in the lipid peroxidation assay. The activity of AQ was lesser when compared with that of ET but greater than PE and DCM. The amount of phenolics and flavonoids were present in higher amounts in ET followed by AQ. Trace amounts of phenolics were detected in PE and DCM, but the amount of flavonoids were below the detection level. The study showed that the antioxidant activity and the concentrations of phenolics and flavonoids are proportionate to each other. Conclusions Ethanolic extract of henna seeds are efficient antioxidants, which can be utilized for further isolation of active compounds and pharmaceutical applications.

466-469

Authors: BR Eesha, Amberkar V Mohanbabu, Kumari K Meena, Sarath babu, M Vijay, M Lalit, R Rajput

Objective To evaluate the hepatoprotective activity of Terminalia paniculata against paracetamol induced hepatic damage in rats. Methods The plant material was shade dried, powdered and extracted with ethanol. Liv 52 and silymarin were used as standard drugs and 2% gum acacia as a control (vehicle). Alteration in the levels of biochemical markers of hepatic damage like AST, ALT, ALP and lipid peroxides were tested, and phytochemical tests were also performed. Results Paracetamol (2 g/kg) increased the serum levels of alanine aminotransfer (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and the lipid peroxides. Treatment of Liv 52, silymarin and ethanolic extract of Terminalia paniculata (200 mg/kg) altered levels of biochemical marker and showed significant hepatoprotective activity. Ethanolic extract revealed the presence of phenolic compound and flavanoids. Our findings suggested that ethanolic bark extract of Terminalia paniculata possessed hepatoprotective activity in a dose dependent manner. Conclusions Terminalia paniculata possesses hepatoprotective activity. It could be an effective and promising preventive agent against PCT induced hepatotoxicity.

475-477

Authors: SC Ohadoma, HU Michael

Objective To investigate the interacting effects of co-administration of methanol leaf extract of Catharanthus roseus (C. roseus) on the hypoglycemic activity of metformin as well as glibenclamide using experimental rats. Methods Phytochemical analysis as well as acute toxicity and lethality (LD50) test were carried out on its methanol leaf extract. The alloxan model for experimental induction of diabetes in rats was employed. Six groups comprising five rats each were used. Groups II, III and IV received 250 mg/kg of extract, 100 mg/kg of metformin and 1 mg/kg of glibenclamide respectively, while V and VI were administered metformin-extract and glibenclamide-extract combinations respectively at doses as above. Group I served as negative control and received only distilled water. All administration was done once daily for seven days. Fasting blood glucose was determined at 2, 12, 24, 72 and 168 h using a glucometer. One-way ANOVA with post-hoc tests was used to assess for significant difference due to administration of drug alone and with co-administration of drug and extract. Results The LD50 was 2 121.32 mg/kg. The phytochemical studies indicated the presence of saponins, tannins, alkaloids, phlotatannins, flavonoids, triterpenoids, reducing sugars, anthraquinones and glycosides. All medicaments significantly reduced blood glucose levels when compared with control alone (P<0.05) with the highest percentage reduction in blood glucose (64.86%) exhibited by metformin-extract combination. Conclusions The leaf extract of C. roseus significantly increases the hypoglycemic effect of metformin.

482-485

Authors: Shengbo Yang, Can Gu, Guiying Zhang, Jian Kang, Haiquan Wen, Qianjin Lu, Jinhua Huang

Objective To explore the molecular mechanisms of antitumor properties of triptolide, a bioactive component isolated from the Chinese herb Tripterygium wolfordii Hook F. Methods Human fibrosarcoma HT-1080 cells were treated with different doses of triptolide for 72 h. Then the expression and activity of matrix metalloproteinase (MMP)-2 and -9 were measured and the invasiveness of triptolide-treated HT-1080 cells was compared with that of anti-MMP-9-treated HT-1080 cells. Results 18 nmol/L triptolide inhibited the gene expression and activity of MMP-9, but not those of MMP-2, in HT-1080 cells. In addition, both 18 nmol/L triptolide and 3 μg/mL anti-MMP-9 significantly reduced the invasive potential of HT-1080 cells, by about 50% and 35%, respectively, compared with the control. Whereas there was no significant difference between the effect of 18 nmol/L triptolide and that of anti-MMP-9 on invasive potential of HT-1080 cells. Conclusions These data suggest that triptolide inhibits tumor cell invasion partly by reducing MMP-9 gene expression and activity.

492-497

Authors: Xinkai Zhao, Qiaoming Ning, Xiaoning Sun, De'an Tian

Objective To investigate the relationship among Pokemon, NF-κ B p65 and Bcl-2 in hepatoma cells. Methods HCC cell HepG2, SMMC7721 and human fetal liver cell line LO2 cells were used, and expression of Pokemon, NF-κ B p65 and Bcl-2 in three cells were detected by real-time PCR and western blot. Then siRNA of Pokemon was applied to inhibit the expression of Pokemon and NF-κ B p65 and apoptotic rate was determined by flow cytometric analysis. Results Expressions of Pokemon, NF-κ B p65 and Bcl-2 in human hepatoma cell HepG2, SMMC7721 expression were significantly higher than those in human embryonic stem cells LO2. siRNA of Pokemon inhibited the expression of Pokemon, NF-κ B p65 and Bcl-2 in liver cancer cells, and significantly increased apoptosis of liver cells. While siRNA of NF-κ B p65 inhibited the expression of NF-κ B p65 and Bcl-2, but Pokemon expression in hepatoma cells had no significant change. Conclusions The proto-oncogene Pokemon can inhibit P14ARF by specific transcription regulation of cell cycle and can induce tumors. In addition, Pokemon can regulate NF-κ B p65 through the expression of apoptosis repressor, and promote the development of liver cancer. It suggests signal network in the liver include the regulation of new non-classical NF-κ B regulatory pathway.