Asian Pacific Journal of Tropical Medicine

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.