Asian Pacific Journal of Tropical Medicine

673-681

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

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

689-692

Authors: KN Sahare, V Singh

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

699-702

Authors: Sirima Kitvatanachai, Pochong Rhongbutsri

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

709-712

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

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

718-722

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

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

728-731

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

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

739-742

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

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

748-752

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

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

757-761

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

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

589-594

Authors: Segun Isaac Oyedeji, Henrietta Oluwatoyin Awobode, Chiaka Anumudu, Jürgen Kun

Objective To characterize the genetic diversity of Plasmodium falciparum (P. falciparum) field isolates in children from Lafia, North-central Nigeria, using the highly polymorphic P. falciparum merozoite surface protein 2 (MSP-2) gene as molecular marker. Methods Three hundred and twenty children were enrolled into the study between 2005 and 2006. These included 140 children who presented with uncomplicated malaria at the Dalhatu Araf Specialist Hospital, Lafia and another 180 children from the study area with asymptomatic infection. DNA was extracted from blood spot on filter paper and MSP-2 genes were genotyped using allele-specific nested PCR in order to analyze the genetic diversity of parasite isolates. Results A total of 31 and 34 distinct MSP-2 alleles were identified in the asymptomatic and uncomplicated malaria groups respectively. No difference was found between the multiplicity of infection in the asymptomatic group and that of the uncomplicated malaria group (P>0.05). However, isolates of the FC27 allele type were dominant in the asymptomatic group whereas isolates of the 3D7 allele type were dominant in the uncomplicated malaria group. Conclusions This study showed a high genetic diversity of P. falciparum isolates in North-central Nigeria and is comparable to reports from similar areas with high malaria transmission intensity.

602-608

Authors: Yogesh Anant Kulkarni, Addepalli Veeranjaneyulu

Objective To study the effects of aqueous extract of Gmelina arborea bark on normoglycemic levels and streptozotocin (STZ) induced diabetes in rats. Methods After single administration of the aqueous extract, plasma glucose level was determined up to 6 h. In subacute study, the aqueous extract was administered for 28 d and plasma glucose level was determined weekly. The diabetes was induced in rats by the intraperitoneal injection of STZ at a dose of 55 mg/kg body weight. The diabetic animals were divided into four groups containing six in each: Group I diabetic control, Group II and III treated with the aqueous extract respectively at a dose of 250 and 500 mg/kg body weight once daily and Group IV treated with glibenclamide at a dose of 0.6 mg/kg body weight once daily. In acute study, the aqueous extract and glibenclamide were administered orally to rats. Plasma glucose levels were determined at 30, 60, 120, 240 and 360 min after the administration of the test samples. To study subacute effects, test samples (the aqueous extract and glibenclamide) were administered for 28 d consecutively. The effects of each test sample on plasma glucose level, body weight as well as food and water intake were also monitored weekly. The oral glucose tolerance test and biochemical indicators were estimated on day 28. Results The aqueous extract did not significantly decrease the plasma glucose level in the normoglycemic rats as shown by the acute and subacute assays. However, after oral administration of the aqueous extract, the plasma glucose level was significantly (P<0.001) decreased in the diabetic rats in the acute study. The long-term administration of the aqueous extract significantly (P<0.001) reduced plasma glucose levels of the diabetic rats. Additionally, the aqueous extract also reduced loss of body weight and significantly decreased food and water intake in the diabetic animals. Nevertheless, no effects on biochemical indicators were observed at the selected doses. Conclusions The aqueous extract of Gmelina arborea bark had antihyperglycemic activity against STZ induced diabetes in rats, after single and subacute oral administration. Moreover, it did not show significant glucose lowering effect in normoglycemic rats.

616-624

Authors: Arzu Birinci Yildirim, Fatma Pehlivan Karakas, Arzu Ucar Turker

Objective To investigate antibacterial and antitumor activities of 51 different extracts prepared with 3 types of solvents (water, ethanol and methanol) of 16 different plant species (Ajuga reptans (A. reptans) L., Phlomis pungens (P. pungens) Willd., Marrubium astracanicum (M. astracanicum) Jacq., Nepeta nuda (N. nuda) L., Stachys annua (S. annua) L., Genista lydia (G. lydia) Boiss., Nuphar lutea (N. lutea) L., Nymphaea alba (N. alba) L., Vinca minor (V. minor) L., Stellaria media (S. media) L., Capsella bursa-pastoris (C. bursa-pastoris) L., Galium spurium (G. spurium) L., Onosma heterophyllum (O. heterophyllum) Griseb., Reseda luteola (R. luteola) L., Viburnum lantana (V. lantana) L. and Mercurialis annua (M. annua) L.) grown in Turkey was conducted. Methods Antibacterial activity was evaluated with 10 bacteria including Streptococcus pyogenes (S. pyogenes), Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis), Escheria coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Salmonella typhimurium (S. typhimurium), Serratia marcescens (S. marcescens), Proteus vulgaris (P. vulgaris), Enterobacter cloacae (E. cloacea), and Klebsiella pneumoniae (K. pneumoniae) by using disc diffusion method. Antitumor activity was evaluated with Agrobacterium tumefaciens (A. tumefaciens)-induced potato disc tumor assay. Results Best antibacterial activity was obtained with ethanolic extract of P. pungens against S. pyogenes. Ethanolic and methanolic extract of N. alba and ethanolic extract of G. lydia also showed strong antibacterial activities. Results indicated that alcoholic extracts especially ethanolic extracts exhibited strong antibacterial activity against both gram-positive and gram-negative bacteria. Best antitumor activity was obtained with methanolic extracts of N. alba and V. lantana (100% tumor inhibition). Ethanolic extract of N. alba, alcoholic extracts of N. lutea, A. reptans and V. minor flowers, methanolic extracts of G. lydia and O. heterophyllum and ethanolic extract of V. lantana and aqueous extract of V. minor leaves exhibited strong tumor inhibitions. Conclusions In near future works, identification of active components can be studied for plant extracts having strong bioactivity.

631-634

Authors: Chintana Chirathaworn, Yong Poovorawan, Somrat Lertmaharit, Norra Wuttirattanakowit

Objective To investigate cytokine profile in patients with chikungunya virus (CHIKV) infection. Methods Twenty eight pairs of serum samples collected from CHIKV infected patients during the outbreak of chikungunya fever in South Thailand in 2008 were obtained. A multiple cytokine assay for detection of 17 cytokines was performed. Results In the acute stage of CHIKV infection, the patients had significantly higher levels of interleukin-6, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, monocyte chemotactic protein 1 and tumor necrosis factor alpha than the control (P<0.001, P=0.023, P=0.015, P<0.001 and P=0.024, respectively). When the disease developed to the recovery stage, the patients had significantly lower levels of interleukin-6, granulocyte-macrophage colony-stimulating factor, monocyte chemotactic protein 1 and macrophage inflammatory protein beta than in the acute stage (P<0.001). Conclusions This study provides additional information that these cytokines could play roles in pathogenesis of CHIKV infection and could be used as disease biomarkers or drug targets.

640-643

Authors: Tie-Jun Li, Jun Cui

Objective To observe cyclooxygenase (COX)-2 expression in normal oral mucosa (NOM), oral lichen planus (OLP) and oral squamous cell carcinoma (OSCC) and explore its significance in the incidence of oral cancer. Methods The immunohistochemical method and RT-PCR method were applied to detect the expression of COX-2 and MMP-7 in 10 cases with NOM, 33 cases of with OLP and 38 cases with OSCC. Results The expression of COX-2 mRNA in OSCC tissues (68.4%, 26/38) was significantly higher than in the OLP (24.2%, 8/33) and NOM (0.0%, 0/10) (P<0.01). The expression of MMP-7 mRNA in OSCC tissues (65.8%, 25/38) was significantly higher than in the OLP (30.3%, 10/33) and NOM (0.0%, 0/10) (P<0.01). The expression of MMP-7 in OLP was significantly higher than in the NOM (P<0.05). There was no significant expression of COX-2 protein in NOM, and the positive rate was 42.4% (14/33) and 89.5% (34/38) in OLP and OSCC group, respectively. The COX-2 expression in cancer tissues was significantly higher than in NOM and OLP (P<0.05). The MMP-7 protein expression in cancer tissues (84.2%, 32/38) was significantly higher than in NOM (10.0%, 1/10) and in OLP (42.4%, 14/33), and the positive rate in OLP was significantly higher than in NOM (P< 0.01). The COX-2 expression was associated with clinical stage (P<0.05), the MMP-7 expression was associated with clinical stage and lymph node metastasis (P<0.05). The expressions of COX-2 and MMP-7 mRNA were positively correlated with OSCC. Conclusions The abnormal expressions of COX-2 and MMP-7 are closely related to the biological behavior of OSCC, the MMP-7 may be induced by COX-2, and further lead to the invasion and metastasis of OSCC.

649-652

Authors: Bo Zhang, Lei Qin, Cui-Jie Zhou, Ye-Lu Liu, Hai-Xin Qian, Song-Bing He

Objective To observe expression of SIRT3 in normal liver tissue, cirrhotic tissue and hepatocellular carcinoma (HCC) tissues, and to explore the significance of SIRT3 in primary HCC. Methods SIRT3 expression was detected in 10 normal cases, 30 cases with, 30 HCC cases by immunohistochemical and Western-blotting method. Results Immunohistochemical assay showed that the SIRT3 positive expression rates were 100.0% (10/10), 96.7% (29/30) and 60.0% (18/30), respectively in normal group, paracancer group and HCC group. And the SIRT3 expression in HCC group was significantly lower than in normal group and paracancer group (P<0.05). Western-blotting showed the SIRT3 expression in cancer tissue was 0.29±0.07, significantly lower than that in paracancer group and normal group (P<0.05). SIRT3 expression was related to the differentiation degree and portal vein tumor thrombus (P<0.05). Conclusions Abnormal expression of SIRT3 is closely related to the biological behavior of primary HCC.

658-662

Authors: Xiao-Lu Liang, Jia-Ying Yuan

Objective To explore the value of B-ultrasound on the evaluation of the effects of traditional Chinese medicine compound of Radix astragali, Salvia miltiorrhiza and Angelica sinensis, and TCM + praziquantel on liver fibrosis in rabbits with schistosomiasis. Methods The hepatic fibrosis model in rabbits with schistosomiasis was established. The experimental animals (24 rabbits) were randomly divided into four groups (group A, B, C and D, n=6). Group A (control group) was only treated by praziquantel; Group B was treated by mixture of Radix astragali and Salvia miltiorrhiza + praziquantel; Group C was treated by mixture of Radix astragali and Angelica sinensis + praziquantel; Group D was treated by mixture of Radix astragali, Salvia miltiorrhiza and Angelica sinensis + praziquantel. Then B-ultrasonogram was used to evaluate the effects. Results Each group showed certain curative effect on liver fibrosis in rabbits with schistosomiasis. The efficacy of group B, C and D was better than group A, and that of group D was the best. The differences in long diameter, thickness diameter, transverse diameter and portal vein inner diameter of liver before and after treatment were statistically significant (P<0.05). The liver function indexes and liver fibrosis indexes were significantly improved after treatment (P<0.05). Conclusions The mixture of Radix astragali, Salvia miltiorrhiza and Angelica sinensis combined with Western medicine treatment can obviously improve the efficacy on liver fibrosis of schistosomiasis.

667-672

Authors: Tarek Tawfik Amin, Hamdan Ibrahim Al-Mohammed, Feroze Kaliyadan, Balghonaim Salah Mohammed

Objective To study epidemiological trends related to cutaneous leishmaniasis (CL) in Al Hassa, an endemic area in Saudi Arabia. Methods This retrospective study included the spatial/temporal analysis of the reported cases of CL using the available surveillance database for the disease at the regional Vector Control Unit, from 2000 to 2010. Results The incidence of CL was declining at a stable rate especially during the last 3 years of the study (2008-2010). An interesting finding was the percentage of expatriates affected was increasing over the last 10 years compared to that of the Saudis. Conclusions A definite declining trend in the incidence of CL was observed in Al Hassa. Further studies are warranted to assess whether special public health measures are needed for better control of CL in expatriate populations in Saudi Arabia.

505-509

Authors: Guadalupe García-Elorriaga, Leila Vera-Ramírez, Guillermo del Rey-Pineda, César González-Bonilla

Objective To determine the polymorphisms of Interleukin-10 (IL-10) (−592, −1082) in pulmonary tuberculosis (PTB) with and without type 2 diabetes (T2D). Methods We studied a Mexican mestizo population of 37 patients with TB in remission (TBr) and 40 with active pulmonary TB (PTB), 21 patients with TB + T2D, 47 blood donors accepted, and 13 healthy health-care workers with tuberculin skin test positive. Determination of IL-10 polymorphisms was performed by real-time Polymerase chain reaction. Results IL-10-592C/A presented in a greater proportion in healthy individuals than in patients with type 2 diabetes and TB in a not quite significant statistically manner. IL-10-1082A/A presented more frequently in the group of patients with both diseases, not being statistically significant in comparison with the group of healthy subjects. Conclusions This study describes two important new findings. First, it reveals that the IL-10 (−592 A/A and −592 C/C) polymorphisms were found in a greater proportion in a group of patients with T2D and TB than in healthy subjects. Second, the study provides evidence that the (−1082 G/G) polymorphism presented with greater frequency in healthy individuals than in patients with both of these diseases.

516-519

Authors: Jin-Hong Luo, Jun Zhou, Yong Gao

Objective To investigate the correlation between periostin and SNCG and esophageal cancer invasion, infiltration and apoptosis. Methods A total of 78 cases esophageal surgical resection specimens were collected, expression of periostin and SNCG in esophageal cancer were detected. Effect of periostin and SNCG in esophageal carcinoma invasion and infiltration was analyzed. Results The upregulated rate of periostin had significant difference in esophageal cancer tissues (39.74%), adjacent tissues (17.86%) and normal tissues (0.00%); The positive expression rates of SNCG had significant difference in esophageal cancer tissues (61.54%), adjacent tissues (32.14%) and normal tissues (1.96%); The upregulated rate of periostin had a significant correlation with lymph node metastasis, adventitia invasion, TNM stage; The positive expression rates of SNCG had a significant correlation with differentiation degree, lymph node metastasis, adventitia invasion, TNM stage; Apoptosis index of the positive of expression of SNCG of esophageal cancer tissue (4.541±2.267) was significantly lower than that of the negative expression (7.316±2.582) (P<0.05). Conclusions SNCG may play an important role in invasion, infiltration and apoptosis of esophageal cancer and serve as target spots in the targeted therapy of esophageal cancer.

532-536

Authors: SN Harsha, KR Anilakumar

Objective To study anxiolytic property of hydro alchohol extract and to estimate polyphenols present in the extract by HPLC. Methods To evaluate anxiolytic property two animal models were used viz. Elevated T maze and hyponeophagia. Diazepam (1 mg/kg body wt.) served as the standard anxiolytic agent for all the tests. The dried extract of the plant leaf in doses of 100, 200 and 400 mg/kg body weight was administered orally to mice for duration of 15 or 30 days and locomotor and anxiolytic activities were performed. Polyphenols was estimated using HPLC. Results The HPLC analysis of the polyphenols revealed the presence chlorogenic acid, vanillin, epicatechin, caffeic acid, rutin hydrate, sinapic acid, quercetin-3-rhamnoside, p-coumeric acid and quercitin. Time spent and number of entries into the open arm was improved in 30 days treated animals than that of 15 days treated groups, 200 and 400 mg/kg body weight treated group showed significant results when comparing with the control group. Conclusions The hydro alcohol extract rich in Polyphenols and other secondary metabolites is a potent anxiolytic agent.

544-547

Authors: Jing Bai, Hui Zhou, Xin-Hong Yang, Hua-Fen Liu, Yan-Yan Meng

Objective To observe the effect on the inhibition of coronary atherosclerosis hardening of the paraoxonase gene (PON-1) which transfected to the rabbit epicardial adipose tissue. Methods Rabbit coronary atherosclerosis model was established by high-fat feeding, liposome-encapsulated recombinant plasmid pEGFP-PON-1 50 μ L was injected to the rabbit pericardial cavity, and was harvested 4 weeks after transfection. Results The epicardial fat transfected PON-1 gene had effect on the high lipid level. It significantly increased expression of PON-1 in peripheral arterial vascular tissue (P <0.05); and significantly reduced total cholesterol and low-density lipoprotein cholesterol levels (P<0.05), and the thickness ratio of coronary artery intima/media (P <0.05). Conclusions The injection of the PON-1 gene in the pericardial cavity can effectively suppress the formation of coronary atherosclerosis.

552-555

Authors: Richard Davis, Trenden Flanigan, Eric Wilson

Objective To determine if passive gravitational sedimentation of blood samples, followed by buffy coat thin smear preparation could increase the sensitivity of malaria diagnosis when compared to conventional thin smear preparation without the additional cost of centrifuges or molecular diagnostics. Methods Blood samples were collected from 205 patients. Each patient sample was analyzed using all three methods of sample preparation. Results Buffy coat analysis of centrifuged blood samples greatly increased the sensitivity of malaria diagnosis when compared to standard thin smear techniques. Sensitivity between mechanically centrifuged samples and gravitationally sedimented samples showed equal improvement in sensitivity when compared to standard thin smear preparation. Conclusions Passive gravitational sedimentation of red blood cells followed by buffy coat analysis dramatically improves the sensitivity of malaria diagnosis without the additional costs associated with centrifugation.

561-569

Authors: JS Ramkissoon, MF Mahomoodally, N Ahmed, AH Subratty

Objective To determine the contribution of total phenolic content (TPC) in glycation inhibitory activity of common tropical medicinal food and spices with potential antioxidative properties. Methods In vitro glucose-bovine serum albumin (BSA) assay was used. Ethanolic extracts of ten common household condiments/herbs (Allium sativum, Zingiber officinale, Thymus vulgaris, Petroselinum crispum, Murraya koenigii Spreng, Mentha piperita L., Curcuma longa L., Allium cepa L., Allium fistulosum and Coriandrum sativum L.) were evaluated for antioxidative activity by 2,2-diphenyl-2-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) and the TPC, flavonoid and tannins content were determined. Results Findings showed good correlation between TPC/DPPH (r = 0.8), TPC/FRAP (r = 0.8), TPC/anti–glycation (r = 0.9), DPPH/anti–glycation (r = 0.6), FRAP/anti–glycation (r = 0.9), Flavonoid/anti–glycation (r = 0.7) and Tannins/anti–glycation (r = 0.8) and relatively fair correlation for TPC/Flavonoids (r = 0.5) and TPC/Tannins (r = 0.5). Results imply that these plants are potential sources of natural antioxidants which have free radical scavenging activity and might be used for reducing oxidative stress. Conclusions The positive glycation inhibitory and antioxidative activities of these tropical herbs suggest a possible role in targeting ageing, diabetic complications and oxidative stress related diseases.

570-573

Authors: Wei-Tao Cheng, Ning Wang

Objective To investigate the correlation between expressions of MMP-2 and NF-κ B in the intracranial aneurysm wall, and explore their role in the mechanism of the occurrence, growth and rupture of intracranial aneurysms. Methods RT-PCR was used to detect the expression of MMP-2 and NF-κ B mRNA of 30 cases of intracranial aneurysm tissue and 10 cases of normal intracranial arterial tissue; Immunohistochemical method was used to detect the expression of MMP-2 and NF-κ B protein. Results The semi-quantitative analysis of MMP-2 and NF-κ B in aneurysms tissues and normal tissues were statistically significant different from each other (P<0 05). Immunohistochemical staining results showed NF-κ B was expressed in different layers. The expression of them were positive in intimal and medial, and the expression sites were located in the nucleus. MMP-2 were expressed in different layers of the aneurysm wall, and the expressions were positive in media and extima. The MMP-2 and NF- κ B-positive expression of aneurysm wall were significantly higher than in normal cerebral arteries (P <0.05). MMP-2 and NF-κ B mRNA expression showed positive correlation in the aneurysm wall tissue (r = 0.689, P = 0.005). Conclusions The expressions of MMP-2 and NF-κ B in the intracranial aneurysm wall tissue were significantly higher than in the normal intracranial arterial tissues. They have a synergistic effect on the formation of intracranial aneurysms.

578-582

Authors: Jia Luo, Yan-Wen Xu, Ming-Fang Zhang, Ling Gao, Cong Fang, Can-Quan Zhou

Objective To explore the effects of different M II stage oocytes zona pellucida birefringence on pregnancy outcome. Methods A total of 46 couples with infertile which induced by single cause received in-vitro fertilization treatment were analyzed retrospectively, and randomly divided into the high zona birefringence (HZB)/HZB group, HZB/low zona birefringence (LZB) group and LZB/LZB group according to different oocytes zona pellucida birefringence. Intracytoplasmic sperm injection outcome was analyzed and compared. Results The proportion of HZB oocytes, implantation rate and the pregnancy rate were decreased in three groups (HZB/HZB group>HZB/LZB group>LZB/LZB group) (P<0.05). But there was no significantly different between the number of oocytes and fertilization rate of these groups (P>0.05). Logistic regression analysis showed that factors affect M II stage oocytes zona pellucida birefringence were age, basal FSH level and the LH level on the day of HCG injection. Age and FSH levels were negatively correlated with the single oocyte zona pellucida birefringence; While the LH level on the day of hCG injection was positively correlated with the single oocyte zona pellucida birefringence. Conclusions The primary influence factors on M II stage oocytes zona pellucida are age, basal FSH level and the LH level on the day of hCG injection. The birefringence value of zona pellucida can affect the pregnancy outcome.

421-425

Authors: Edward O Salami, Raymond I Ozolua, Stephen O Okpo, Gerald I Eze, Dickson O Uwaya

Objective To evaluate the antiasthmatic and antitussive properties of the aqueous leaf extract of Bryophyllum pinnatum (B. pinnatum) (BP) Lam. Methods Ovalbumin-sensitized guinea pigs which were treated with BP for 21 consecutive days were exposed to 0.2% histamine aerosol in a glass chamber. Mucus viscosity, white blood cell and lymphocyte counts and tracheal wall morphometry were measured. Bouts of cough were counted pre and post acute exposure of extract-treated (× 7 d) guinea pigs to 7.5% citric acid aerosol in a chamber. Phenol red expectoration was estimated in mice after 7 d of daily administration of BP. Results Doses of 200 and 400 mg/kg/day (×21 d) BP significantly increased the time for guinea pigs to experience preconvulsive dyspnoea. BP and salbutamol (0.5 mg/kg/day ×21 d) reduced mucus viscosity in the sensitized group to values comparable with controls. White blood cell, lymphocyte counts and tracheal morphometry were not significantly altered. Both doses of BP also significantly reduced the bouts of cough but only 400 mg/kg/day significantly inhibited the amount of phenol red secreted. Conclusions BP has demonstrated antiasthmatic and antitussive properties in these rodent models. These properties may underscore its use in Nigerian ethnomedicine.

433-437

Authors: Imran Kazmi, Muhammad Afzal, Babar Ali, Zoheir A. Damanhouri, Aftab Ahmaol, Firoz Anwar

Objective To investigate the anxiolytic activity of newly isolated compound by our lab called ursolic acid stearoyl glucoside (UASG) from the leaves ofLantana camara (L. camara). Methods Column chromatography was used to isolate UASG. Anxiolytic potential was experimentally proved and demonstrated through Elevated plus-maze, Open field and light and dark test. Results The UASG showed marked increased in time spent (%) and number of frequent movements made by animals in open arm of elevated plus-maze apparatus. In light and dark model, UASG produced marked increase in time spent by animal, number of crossing and reduced duration of immobility in light box. Conclusions UASG showed significant increase in number of rearing, assisted rearing and number of square crossed in open field established test model. UASG showed its anxiolytic effect in dose dependent manner.

443-448

Authors: Pabitra Saha, Arindam Naskar, Swagata Ganguly, Sonali Das, Subhasish K Guha, Asit Biswas, Dilip K Bera, Pratip K Kundu, Madhusudan Das, Shrabanee Mullick, Krishnangshu Ray, Ardhendu K Maji

Objective To study the in vivo efficacy of these two ACTs in the treatment ofPlasmodium falciparum (P. falciparum malaria) in Kolkata and to determine the prevalence of mutant S769N codon of thePfATPase6 gene among field isolates ofP. falciparum collected from the study area. Methods A total of 207P. falciparum positive cases were enrolled randomly in two study arms and followed up for 42 days as per WHO (2009) protocol. A portion ofPfATPase6 gene spanning codon S769N was amplified and sequenced by direct sequencing method. Results It was observed that the efficacy of both the ACT regimens were highly effective in the study area and no mutant S769N was detected from any isolate. Conclusions The used, combination AS+SP is effective and the other combination AM+LF might be an alternative, if needed.

453-456

Authors: Reza Mirnejad, Mozafar mohamadi, Vahbeh Piranfar, Seied Mojtaba Mortazavi, Reza Kachuei

Objective To design a duplex PCR for rapid and simultaneous detection of Brucella species. in human blood samples. Methods Fifty-two peripheral bloods samples were collected from suspicious patients with brucellosis. Following DNA extraction, PCR assay were performed, using three primers that could simultaneously identify and differentiate three major species of pathogenic Brucella in humans and animals. Results Of the 52 peripheral bloods samples tested, 25 sample (48%) showed positive reactions in PCR. Twelve samples were positive for Brucella abortus 39 (B. abortus 39) (23%), 13 for Brucella melitensis 39 (B. melitensis 39) (25%) and 0 for Brucella ovis 39 (B. ovis 39) (0%). Conclusions This work de=monstrates that in case where specific primers were utilized, duplex PCR has proved to be a simple, fast, and relatively inexpensive method for simultaneous detection of important species of Brucella in clinical samples.

462-466

Authors: Md. Shahin Mia, Rawshan Ara Begum, A.C. Er, Raja Datuk Zaharaton Raja Zainal Abidin, Joy Jacqueline Pereira

Objective To analyze trends of dengue incidences and deaths in Malaysia from 2000 to 2010 as well as the predominant dengue virus serotypes during the last decade. Methods We used the national data on annual reported cases, deaths, incidence rate, mortality rate, and case fatality rate of dengue fever (DF) and dengue hemorrhagic fever (DHF) as well as dengue virus serotypes prevalent in Malaysia during the last decade. Trend/ regression lines were fitted to investigate the trend of dengue incidences and deaths due to the disease for a 11-year period (2000-2010). For the distribution of national incidence rate, mortality rate, and case fatality rate of DF and DHF, descriptive statistics using mean and 95% confidence intervals (CI 39) for means, and range were applied. Results The number of dengue cases and number of deaths have increased, on average, by 14% and 8% per year respectively. The average annual incidence rate of DF per 100 000 populations was higher as compared to that of DHF. Conversely, the yearly mean mortality rate of DHF per 100 000 populations was greater than that of DF. The simultaneous circulation of all four dengue serotypes has been found in Malaysia. But a particular dengue virus serotype predominates for at least two years before it becomes replaced by another serotype. Conclusions The dengue situation in Malaysia has worsened with an increasing number of reported cases and deaths during the last decade. The increasing trend of dengue highlights the need for a more systematic surveillance and reporting of the disease.

473-480

Authors: Lei Yu, Yang-De Zhang, Jun Zhou, De-Ming Yao, Xiang Li

Objective To indentify target genes of transcription factor CCAAT enhancer-binding protein β (CEBPB) in acute promyelocytic leukemia cells induced by all-trans retinoic acid. Methods A new strategy for high-throughput identification of direct target genes was established by combining chromatin immunoprecipitation (ChIP) with in vitro selection. Then, 106 potential CEBPB binding fragments from the genome of the all-trans retinoic acid (ATRA)-treated NB4 cells were identified. Results Of them, 82 were mapped in proximity to known or previously predicted genes; 7 were randomly picked up for further confirmation by ChIP-PCR and 3 genes (GALM, ITPR2 and ORM2) were found to be specifically up-regulated in the ATRA-treated NB4 cells, indicating that they might be the down-stream target genes of ATRA. Conclusions Our results provided new insight into the mechanisms of ATRA-induced granulocytic differentiation.

485-488

Authors: Jiang-Hua Zhong, Shi-Juan Lu, Mo-Shui Chen, Zi-Bin Chen, Liu Wang, Ping-Sheng Wu

Objective To observe effects of hypokalemia on transmural heterogeneity of ventricular repolarization in left ventricular myocardium of rabbit, and explore the role of hypokalemia in malignant ventricular arrhythmia (MVA). Methods A total of 20 rabbits were randomly divided into control group and hypokalemic group. Isolated hearts in the control group were simply perfused with modified Tyrode's solution, and were perfused with hypokalemic Tyrode's solution in hypokalemic group. Ventricular fibrillation threshold (VFT), 90% monophasic action potential repolarization duration (APD90) of subepicardial, midmyocardial and subendocardial myocardium, transmural dispersion of repolarization (TDR) and C×43 protein expression in three layers of myocardium were measured in both groups. Results VFT in the control group and the hypokalemic group were (13.40±2.95) V, and (7.00±1.49) V, respectively. There was a significant difference between two groups (P<0.01). APD90 of three myocardial layers in the hypokalemic group were significantly prolonged than those in the control group (P<0.01). ΔAPD90 in the hypokalemic group and the control group were (38.10±10.29) ms and (23.70±5.68) ms, and TDR were (52.90±14.55) ms and (36.10±12.44) ms, respectively. ΔAPD90 and TDR in the hypokalemic group were significantly higher than those in the control group (P<0.05), and the increase in APD90 of midmyocardium was more significant in the hypokalemic group. Cx43 protein expression of all three myocardial layers were decreased significantly in the hypokalemic group (P<0.01), and ΔCx43 was significantly increased (P<0.05). Reduction of Cx43 protein expression was more significant in the midmyocardium. Conclusions Hypokalemic can increase transmural heterogeneity of C×43 expression and repolarization in left ventricular myocardium of rabbit, and decrease VFT and can induce MVA more easily.

493-496

Authors: Ding-Zhu Shen, San-Li Xin, Chuan Chen, Te Liu

Objective To study the effect of atorvastatin on atherosclerotic rabbits. Methods A total of 60 New Zealand male rabbits were randomly divided into the normal group, model group and atorvastatin group. The replication rabbit atherosclerotic model with immune injury combined with a high fat diet feeding was used. All rabbits were sacrificed after 3 months. TLR4 and NF-κB p65 were observed by HE staining, immunohistochemistry and western blotting. Results The expression of TLR4, NF-κB p65 were significantly increased in the model group compared with the normal group. The expression of TLR4 and NF-κB p65 decreased significantly in the atorvastatin group, and there was no difference compared with the normal group. Conclusions The effect of atorvastatin on atherosclerosis may be achieved by the inhibition of the expression of TLR4 and NF-κB p65.

500-501

Authors: Yingjuan Mok, Jessica Quah, Chuin Siau

Severe rhabdomyolysis is an uncommon but potentially fatal complication of dengue fever that is not well characterised and may be underreported. With the resurgence and continued rise of dengue cases worldwide, physicians must be aware of the less common but serious complications of dengue. Here, we report a patient who presented with severe rhabdomyolysis secondary to dengue fever with a serum creatine kinase of 742 900 U/L.

504

Authors: Hafiz Azhar Ali Khan, Waseem Akram

None

337-345

Authors: VGM Naidu, Uma Mahesh Bandari, Ashwini Kumar Giddam, Kuppan Rajendran Dinesh Babu, Jian Ding, K Suresh Babu, B Ramesh, Rajeswara Rao Pragada, P Gopalakrishnakone

Objective To evaluate the anti-proliferative and apoptogenic activity of ethyl acetate extract from the leaves of Memecylon edule (EtAc-LME) in MKN-74, NUGC gastric cancer cells and non cancerous gastric mucous cells (GES-1), and to explore the mechanism of EtAc-LME induced apoptosis. Methods The mechanism of EtAc-LME induced apoptosis was explored by analysing the activation of pro-caspases, PARP cleavage, expression of cytochrome-c (Cyt-c) was determined by western blotting, mRNA expression of Bcl-2, Bax by RT-PCR, loss of mitochondrial potential using DiOC6 dye, annexin binding assay and its influence on cell cycle arrest by flow cytometry. Results The results indicated that EtAc-LME inhibited the gastric cancer cell growth in dose-dependent manner and cytotoxicity was more towards the gastric cancer cells (NUGC and MKN-74) compared to normal gastric cells (GES-1), suggesting more specific cytotoxicity to the malignant cells. Over expression of Cyt-c and subsequent activation of caspases-3 and down regulation of Bcl-2 and loss in mitochondrial potential in EtAc-LME treated MKN-74 and NUGC cells suggested that EtAc-LME induced apoptosis by mitochondrial dependent pathway. Conclusions The present findings suggest that ethyl acetate extract of Memecylon edule induces apoptosis selectively in gastric cancer cells emphasizing the importance of this traditional medicine for its potential in the treatment of gastric cancer.

352-359

Authors: Muhammad Kamruzzaman, S.M. Nayeemul Bari, Shah M. Faruque

Objective To investigate in vitro and in vivo antibacterial potentials of Vitex negundo (V. negundo) leaf extracts against diverse enteric pathogens. Methods Water and methanol extracts of V. negundo leaves were evaluated against enteric bacterial pathogens by using standard disc diffusion, viable bacterial cell count methods, determination of minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC). Results Methanol extract of V. negundo leaves showed potent antibacterial activity (inhibition zone: 9.9–22.6 mm, MIC: 200–3200 μg/mL, MBC: 200–6400 μg/mL) against all the pathogenic enteric bacteria (Vibrio cholerae, Vibrio parahaemolyticus, Vibrio mimicus, Echerichia coli, Shigella spps., and Aeromonas spps) tested. Methanol extract of V. negundo leaves showed potent bactericidal activity both in vitro laboratory conditions (MBC, 200-400 μg/mL) and in the intestinal environment (Dose, 1-2 mg/mL) of infant mice against pathogenic Vibrio cholerae, the major causative agent of cholera. Furthermore, assays using the mice cholera model showed that V. negundo methanol extract can protect mice from Vibrio cholerae infection and significantly decrease the mortality rate (P<0.0001). Conclusions For the first time we showed that methanol extract of V. negundo leaves exhibited strong vibriocidal activity both in vitro and in vivo conditions. Therefore, it will be useful to identify and isolate the active compounds of this extract that could be a good alternative of antibiotics to treat cholera.

366-371

Authors: Rambir Singh, Shariq Hussain, Rajesh Verma, Poonam Sharma

Objective To find out the anti-mycobacterial potential of Cassia sophera (C. sophera), Urtica dioica (U. dioica), Momordica dioica, Tribulus terrestris and Coccinia indica plants against multi-drug resistant (MDR) strain of Mycobacterium tuberculosis (M. tuberculosis). Methods Plant materials were extracted successively with solvents of increasing polarity. Solvent extracts were screened for anti-mycobacterial activity against fast growing, non-pathogenic mycobacterium strain, Mycobacterium semegmatis, by disk diffusion method. The active extracts were tested against MDR and clinical isolates of M. tuberculosis by absolute concentration and proportion methods. The active extracts were subjected to bio-autoassay on TLC followed by silica column chromatography for isolation of potential drug leads. Results Hexane extract of U. dioica (HEUD) and methanol extract of C. sophera (MECS) produced inhibition zone of 20 mm in disc diffusion assay and MIC of 250 and 125 μ g/mL respectively in broth dilution assay against Mycobacterium semegmatis. Semipurified fraction F2 from MECS produced 86% inhibition against clinical isolate and 60% inhibition against MDR strain of M. tuberculosis. F18 from HEUD produced 81% inhibition against clinical isolate and 60% inhibition against MDR strain of M. tuberculosis. Phytochemical analysis indicated that anti-mycobacterial activity of MECS may be due to presence of alkaloids or flavonoids and that of HEUD due to terpenoids. Conclusions C. sophera and U. dioica plant extracts exhibited promising anti-mycobacterial activity against MDR strain of M. tuberculosis. This is the first report of anti-mycobacterial activity form C. sophera. This study showed possibility of purifying novel anti-mycobacterial compound(s) from C. sophera and U. dioica.

379-382

Authors: Li-Feng Qin, Wan-Chun Wang, Han Fang, Xin-Zhan Mao, Guo-Liang Huang, You Chen, Hou-De Zhou, Yi Shen, Li-Hua Qin, Dan Peng

Objective To explore the significance of osteopontin and nuclear factorκB (NF-κB) expression in patients with knee osteoarthritis. Methods RT-PCR and enzyme-linked immunosorbent assay were used to measure the Osteopontin (OPN) and NF-κB concentration of knee joint synovial fluid of patients with knee osteoarthritis and trauma fractures, and analyze the relationship between the expressiones of them. Results OPN and NF-κB expression at the mRNA and protein levels of patients with knee osteoarthritis were significantly higher than the control group. the result showed statistical significance (P<0.05). There was a positive correlation between the OPN levels in synovial fluid of patients with knee osteoarthritis and NF-κB expression levels (P<0.05). Conclusions The high expression of OPN and NF-κB are closely related to occurrence and development of knee osteoarthritis.

387-394

Authors: Ke Tao, Hong-Tao Wang, Bi Chen, Bo-Tao Wang, Zhi-Yuan Li, Xiong-Xiang Zhu, Chao-Wu Tang, Da-Hai Hu

Objective To investigate the effect and the relevant potential mechanism of nonpeptide neurokinin 1 (NK1) receptor antagonist L-703,606 in the edema formation after burn injury. Method L-703,606 treatment was performed in Sprague-Dawley (SD) rats at early stage after deep partial-thickness skin scalding. One hundred and fifty two adult male SD rats were used in the study and randomly divided into sham scald (SS, n=8), scald control (SC, n=48), and L-703,606 treatment (LT, n=48) groups. The rats in SC and LT groups were subjected to 20% total body surface area (TBSA) deep partial-thickness skin scalding. Modified Evans blue extravasation, tracing electron microscopy by lanthanum nitrate and mean water content assay were employed to observe and detect the changes of vascular permeability, ultrastructure and edema formation in adjacent tissue to the wounds and in the jejuna of rats at early stage (72 h) after scald. Results The pathological increase of vascular permeability in the periwound tissue and jejunum of rats in LT group were significantly lower than that in SC group (P<0.01), and recuperated earlier. Meanwhile, the changes of water contents of corresponding tissues in LT group were lighter than those in SC group (P<0.01). The ultrastructural changes of the microvessels in the peri-wound tissue of LT group showed that the junctions between microvascular endothelium cells were more narrow than those of SC group, moreover, and the number of opening and the engorgement and cavitation of the vascular endothelium cells decreased, the areosis and edema in perivascular tissue lightened, and the precipitation of the high eletron density lanthanum tracing agent in the interspace of the tissue decreased significantly in LT group. Conclusions It is concluded that nonpeptide NK1-receptor antagonist L-703,606 could lighten the vascular permeability and edema formation in the periwound tissue and jejunum, and accelerate the normalization process of pathological changes in the tissues of rats after scald.

400-403

Authors: Zhong Li

Objective To discuss the value of lymph node mapping in rabbit liver cancer with nanocarbon and methylene blue injecta. Methods Rabbit liver cancer model was established by transplanting VX2 cells with laparotomy in celiac planting method. Twenty Japan white rabbits were divided into two groups randomly. Each group had 10 rabbits. Lymph node mapping in two groups rabbit liver cancer were observed. Two groups rabbit liver cancer and local lymph nodes were removed. The number and location of local lymph nodes were recorded, and then the samples were obtained from both groups. Results The lymph nodes dyed time was (100.50±29.92) s in nanocarbon group, and (11.20±4.18) s in methylene blue group with statistical significance between two groups (P=0.000). In the comparison of lymph node fading time, nanocarbon group was (2.22±0.74) h, methylene blue group was (1.63±0.54) h, nanocarbon group was longer than the methylene blue group, but without statistical significance (P=0.058). The accuracy was 87.5% (35/40) in methylene blue group, while, the nanocarbon group was 87.2% (34/39), with statistical significance (P=1.000). Conclusions Experimental results show that application of nanocarbon injection and methylene blue injection during resection of liver cancer and local lymph nodes in rabbit liver cancer model has obvious tracer function in liver cancer and lymphatic drainage. It can reduce the complexity and risk of the operation, and avoid the blindness in the process of traditional lymph node dissection surgery. Besides, they can effectively reduce the number of residual lymph nodes after operation. It can achieve the lymph node dissection more thoroughly, promptly, easily and safely.

407-409

Authors: Xiao-Feng Li, Lin Ma, Jing Lu, Li-Xia Kong, Xiao-Hua Long, Su-Huan Liao, Bao-Rong Chi

Objective To discuss effect of ionizing radiation on transcription of colorectal cancer multidrug resistance (MDR) 1 gene of HCT-8 cells. Methods Total RNA was extracted by guanidine thiocyanate one-step method. Northern blot was applied to detect transcription level of MDR1 gene. The expression of P-gp protein was detected by flow cytometry. Results The expression of MDR1 of normal colorectal cancer HCT-8 cells was low. It was increased by 8.35 times under stimulus with 2 Gy. When treated with low doses in advance, high expressed MDR was decreased significantly under 0.05, 0.1 Gy, which was 69.00%, 62.89% in 2 Gy group and 5.77 times, 5.25 times in sham irradiation group. No obvious difference was detected between (0.2+2) Gy group and 2 Gy group. Compared with sham irradiation group, the percentage of P-gp positive cells after radiation of a high 2 Gy dose was increased significantly (P<0.01). When treated with high radiation dose following low radiation dose (0.05 Gy, 0.1 Gy) in advance, the percentage of P-gp positive cells were also increased significantly. The percentage of P-gp positive cells were increased obviously in 0.2 Gy and 2 Gy groups. Compared with simple high radiation 2 Gy group, the percentage of P-gp positive cells was decreased significantly (P<0.05). Conclusions Low radiation dose can reverse multidrug resistance of colorectal cancer cells caused by high radiation dose.

416-418

Authors: Hari Krishan Aggarwal, Deepak Jain, Vipin Kaverappa, Promil Jain, Ashwani Kumar, Sachin Yadav

In tropical countries like India, malaria has been one of the most common parasitic illnesses leading to frequent hospitalization and causing major economic burden among the masses. Although Plasmodium vivax infection is considered to be benign, in contrast to Plasmodium falciparum infection which is notorious for its severe splenic complications can occur frequently. Splenomegaly tends not to receive special attention, as it is not usually accompanied by any symptoms and can be gradually resolved via standard antimalarial therapy. Splenic infarction, although rarely attributable to malaria in an endemic region with high parasitemia, can be a rare presentation of this disease entity.