Asian Pacific Journal of Tropical Medicine

1-10

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

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

20-26

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

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

37-42

Authors: Jude C Ikewuchi

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

49-52

Authors: Hong-Tao Liu, Chun-Lei Lu

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

57-60

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

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

68-72

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

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

76-78

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

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

82-84

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

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