Asian Pacific Journal of Tropical Medicine

Volume 9,Issue 3,2016 Table of Contents

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  • 1  Historic accounts of Mansonella parasitaemias in the South Pacific and their relevance to lymphatic filariasis elimination efforts today
    J. Lee Crainey Túllio Romo Ribeiro da Silva Sérgio Luiz Bessa Luz
    2016, 9(3):201-207. DOI: 10.1016/j.apjtm.2016.01.040
    [Abstract](63) [HTML](0) [PDF 695.76 K](269)
    There are two species of filarial parasites with sheathless microfilariae known to commonly cause parasitaemias in humans: Mansonella perstans and Mansonella ozzardi. In most contemporary accounts of the distribution of these parasites, neither is usually considered to occur anywhere in the Eastern Hemisphere. However, Sir Patrick Manson, who first described both parasite species, recorded the existence of sheathless sharp-tailed Mansonella ozzardilike parasites occurring in the blood of natives from New Guinea in each and every version of his manual for tropical disease that he wrote before his death in 1922. Manson's reports were based on his own identifications and were made from at least two independent blood sample collections that were taken from the island. Pacific region Mansonella perstans parasitaemias were also later (in 1923) reported to occur in New Guinea and once before this (in 1905) in Fiji. Although Mansonella-parasitaemias are generally regarded as benign, they are thought to be of public health importance because they can affect the epidemiological monitoring of other filarial diseases. In this article, we reviewed the historic literature concerning Pacific-origin Mansonella-parasitaemias in an attempt to explain how, despite repeated reports of Pacific-region Mansonella-parasitaemias, by as early as the 1970s, the WHO had arrived at the present-day view that Wuchereria bancrofti is the only cause of filarial parasitaemias in Papua New Guinea. We have also evaluated the evidence supporting the contemporary existence of Pacific-area parasitaemia-causing Mansonella parasites and assessed the relevance such parasites could have for present-day lymphatic filariasis elimination efforts in the region.
    2  Antihydatic and immunomodulatory effects of Punica granatum peel aqueous extract in a murine model of echinococcosis
    Moussa Labsi Lila Khelifi Dalila Mezioug Imene Soufli Chafia Touil-Boukoffa
    2016, 9(3):208-216. DOI: 10.1016/j.apjtm.2016.01.038
    [Abstract](51) [HTML](0) [PDF 1.01 M](322)
    Objective: To investigate the effect of pomegranate peel aqueous extract (PGE) on the development of secondary experimental echinococcosis and on the viability of Echinococcus granulosus protoscoleces, and the immunomodulatory properties of PGE. Methods: Swiss mice were inoculated intraperitoneally with viable protoscoleces. Then, PGE was orally administered daily during cystic echinococcosis development. Cyst development and hepatic damage were macroscopically and histologically analyzed. The production of nitric oxide and TNF-α was assessed in plasma and the hepatic expression of iNOS, TNF-α, NF-κB and CD68 was examined. Moreover, protoscoleces were cultured and treated with different concentrations of PGE. Results: It was observed that in vitro treatment of protoscoleces caused a significant decrease in viability in a PGE-dose-dependent manner. In vivo, after treatment of cystic echinococcosis infected mice with PGE, a significant decrease in nitric oxide levels (P<0.000 1) and TNF-α levels (P<0.001) was observed. This decline was strongly related to the inhibition of cyst development (rate of hydatid cyst growth inhibition=63.08%) and a decrease in CD68 expression in both the pericystic layer of hepatic hydatid cysts and liver tissue (P<0.000 1). A significant diminution of iNOS, TNF-α and NF-κB expression was also observed in liver tissue of treated mice (P<0.000 1). Conclusions: Our results indicate an antihydatic scolicidal effect and immunomodulatory properties of PGE, suggesting its potential therapeutic role against Echinococcus granulosus infection.
    3  Hepatoprotective effects of Nigella sativa seed extract against acetaminophen-induced oxidative stress
    Gareeballa Osman Adam Md. Mahbubur Rahman Sei-Jin Lee Gi-Beum Kim Hyung-Sub Kang Jin-Shang Kim Shang-Jin Kim
    2016, 9(3):217-222. DOI: 10.1016/j.apjtm.2016.01.039
    [Abstract](92) [HTML](0) [PDF 862.54 K](240)
    Objective: To investigate the protective effects of Nigella sativa seed extract (NSSE) against acetaminophen (APAP)-induced hepatotoxicity in TIB-73 cells and rats. Methods: Toxicity in TIB-73 cells was induced with 10 μmol/L APAP and the protective effects of NSSE were evaluated at 25, 50, 75, 100 μg/mL. For in vivo examination, a total of 30 rats were equally divided into five experimental groups; normal control (vehicle), APAP (800 mg/kg body weight single IP injection) as a hepatotoxic control, and three APAP and NS pretreated (2 weeks) groups (APAP+NSSE 100 mg; APAP+NSSE 300 mg and APAP+NSSE 900 mg/kg). Results: TIB-73 cell viability was drastically decreased by (49.0±1.9)% after the 10 μmol/L APAP treatment, which also increased reactive oxygen species production. Co-treatment with NSSE at 25, 50, 75, and 100 μg/mL significantly improved cell viability and suppressed reactive oxygen species generation. In vivo, the APAP induced alterations in blood lactate levels, pH, anionic gap, and ion levels (HCO3 , Mg2+ and K+), which tended to normalize with the NSSE pretreatment. The NSSE also significantly decreased elevated serum levels of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and alkaline phosphatase induced by APAP, which correlated with decreased levels of hepatic lipid peroxidation (malondialdehyde), increased superoxide dismutase levels, and reduced glutathione concentrations. Improved hepatic histology was also found in the treatment groups other than APAP group. Conclusions: The in vitro and in vivo findings of this study demonstrated that the NSSE has protective effects against APAP-induced hepatotoxicity and metabolic disturbances by improving antioxidant activities and suppressing both lipid peroxidation and ROS generation.
    4  Achillea fragrantissima, rich in flavonoids and tannins, potentiates the activity of diminazine aceturate against Trypanosoma evansi in rats
    Ibrahim M. El-Ashmawy Naser A. Al-Wabel Aida E. Bayad
    2016, 9(3):223-229. DOI: 10.1016/j.apjtm.2016.01.032
    [Abstract](55) [HTML](0) [PDF 698.48 K](284)
    Objective: To evaluate activity of methanol extract of Achillea fragrantissima (meth) (A. fragrantissima) alone or in combination with diminazine aceturate (DA) against Trypanosoma evansi in experimentally infected rats. Methods: Sixty adult male Wister albino rats were divided equally into 6 groups (A-F). Rats in groups A-E were experimentally infected with T. evansi and those in group F were uninfected. The groups were treated respectively as follows: group A- with 3.5 mg/kg DA; group B- with 1 000 mg/kg meth A. fragrantissima; group C-3.5 mg/kg DA plus 500 mg/kg meth A. fragrantissima; group D-3.5 mg/kg DA plus 1 000 mg/ kg meth A. fragrantissima. Group E was left untreated. Parasitaemia, survivability, packed cell volume, hemoglobin concentration, total leucocytes count, lymphocyte count, and serum malondialdehyde and reduced glutathione (GSH) levels were estimated. Phytochemical screening of meth A. fragrantissima was also performed. Results: The phytochemical analysis of the meth A. fragrantissima indicated a higher content from polyphenolic tannins and non tannins and flavonoids. The efficacy percentage against trypanosomiasis in groups A to E was respectively as follows 80, 40, 90, 100, 0. The administration of meth-A. fragrantissima (1 000 mg/kg b.wt.) produced a moderate efficacy against trypanosomiasis. Untreated rats in group E died between 25 and 30 d post infection. The rats given DA and meth A. fragrantissima combinations (C and D) showed faster and higher recovery rates than the uninfected control and groups A and B. The initial reduction in packed cell volume, hemoglobin, total leucocytes count, increases in serum malondialdehyde and decreases in GSH levels were reversed by the treatments. Conclusions: The administration of the methanol extracts of A. fragrantissima and DA combination therapy was more effective than each product alone in the treatment of rats infected with Trypanosoma evansi and further studies are required to isolate more active ingredients.
    5  Detection of coat protein gene of nervous necrosis virus using loop-mediated isothermal amplification
    Jinik Hwang Sung-Suk Suh Mirye Park Myung-Joo Oh Jong-Oh Kim Sukchan Lee Taek-Kyun Lee
    2016, 9(3):230-235. DOI: 10.1016/j.apjtm.2016.01.035
    [Abstract](68) [HTML](0) [PDF 933.44 K](281)
    Objective: To establish a novel and highly specific loop-mediated isothermal amplification (LAMP) assay for the identification of nervous necrosis virus (NNV) infection. Methods: A set of synthesized primers was used to match the sequences of a specific region of the nnv gene from the National Center for Biotechnology Information database, not originating from NNV-infected fish, the efficiency and specificity of LAMP were measured dependent on the concentration of DNA polymerase and the reaction temperature and time. In addition, to determine species-specific LAMP primers, cross reactivity testing was applied to the reaction between NVV and other virus families including viral hemorrhagic septicemia virus and marine birnavirus. Results: The optimized LAMP reaction carried out at 64 ℃ for 60 min, and above 4 U Bst DNA polymerase. The sensitivity of LAMP for the detection of nnv was thus about 10 times greater than the sensitivity of polymerase chain reaction. The LAMP assay primers were specific for the detection NNV infection in Epinephelus septemfasciatus. Conclusions: The development of LAMP primers based on genetic information from a public database, not virus-infected samples, may provide a very simple and convenient method to identify viral infection in aquatic organisms.
    6  Pharmacodynamics of aminoglycosides and tetracycline derivatives against Japanese encephalitis virus
    Rashmee Topno Siraj Ahmed Khan Pritom Chowdhury Jagadish Mahanta
    2016, 9(3):236-240. DOI: 10.1016/j.apjtm.2016.01.033
    [Abstract](80) [HTML](0) [PDF 911.04 K](232)
    Objective: To explore the antiviral activity of antibiotic compounds, mainly aminoglycosides and tetracyclines against Japanese encephalitis virus (JEV) induced infection in vitro.Methods: Antiviral activity were evaluated against JEV using cytopathic effect inhibition assay, virus yield reduction assay, caspase 3 level, extracellular viral detection by antigen capture ELISA and viral RNA levels. Results: JEV induced cytopathic effect along with reduction of viral progeny plaque formation indicated antiviral potential of the compounds suggesting that antibiotics had broad spectrum activity. Doxycycline and kanamycin administration in dose dependent manner declined viral RNA replication. Conclusions: The present study shows kanamycin and doxycycline can affect virion structure and alter replication causing inhibition of JEV induced pathogenesis in vitro.
    7  Relationship between CD14-159C/T gene polymorphism and acute brucellosis risk
    Mehdi Moghadampour Ebrahim Eskandari-Nasab Fatemeh Shabani
    2016, 9(3):241-245. DOI: 10.1016/j.apjtm.2016.01.036
    [Abstract](44) [HTML](0) [PDF 693.02 K](232)
    Objective: To investigate the association between the cluster of differentiation 14 (CD14)159C/T (rs2569190) gene polymorphism and susceptibility to acute brucellosis in an Iranian population. Methods: The study included 153 Iranian patients with active brucellosis and 128 healthy individuals as the control group. Genotyping of the CD14 variant was performed using an amplification refractory mutation system-polymerase chain reaction method. Results: The prevalence of CD14-159 TT and CT genotypes were associated with increased risk of brucellosis [odds ratio (OR)=1.993, 95% confidence interval (95% CI)=1.07-3.71, P=0.03 for CT; OR=3.869, 95% CI=1.91-7.84, P=0.01 for TT genotype. Additionally, the minor allele (T) was significantly more frequently present in brucellosis patients than in controls (61% vs. 45%, respectively), and was a risk factor for brucellosis (OR=3.058, 95% CI=1.507-6.315, P=0.01). Conclusions: The findings provid suggestive evidence of association of the CD14159C/T gene polymorphism with susceptibility to acute brucellosis in the Iranian population.
    8  Echovirus serotypes circulating in Malaysia from 2002 to 2013
    Jeyanthi Suppiah TS Saraswathy Subramaniam Amry Khursany Ismail Apandi Yusof Zainah Sa’at
    2016, 9(3):246-249. DOI: 10.1016/j.apjtm.2016.01.037
    [Abstract](44) [HTML](0) [PDF 697.59 K](244)
    Objective: To identify the circulating serotypes of human echovirus in Malaysia from 2002 to 2013. Methods: A total of 31 retrospective samples from non-polio acute flacid paralysis, hand-food-and-mouth disease, viral meningitis and enterovirus cases were subjected to amplification of partial VP1 gene by RT-PCR. Results: Sequencing and phylogenetic analysis of the partial sequences identified presence of human echovirus and human coxsackie viruses. It was found that echovirus 11 was the commonly circulating serotype followed by echovirus 6, echovirus 7, echovirus 3, echovirus 9, echovirus 30 and echovirus 1 in decreasing order. Additionally two types of human coxsackie virus isolates were detected which were coxsackie A24 and B3. Conclusions: From the findings, there is a possibility that echovirus 11 is the predominant serotype among Malaysian patients with echovirus infection. However, a larger sample size will yield a more confident result to support this evidence.
    9  Characteristics, clinical outcomes and factors influencing mortality of patients with melioidosis in southern Thailand: a 10-year retrospective study
    Chaitong Churuangsuk Sarunyou Chusri Thanaporn Hortiwakul Boonsri Charernmak Kachornsakdi Silpapojakul
    2016, 9(3):250-255. DOI: 10.1016/j.apjtm.2016.01.034
    [Abstract](62) [HTML](0) [PDF 656.41 K](240)
    Objective: To study characteristics, clinical outcomes and factors influencing mortality of patients afflicted with melioidosis. Methods: A total of 134 patients were retrospectively analyzed with a microbiologically-confirmed diagnosis of melioidosis, during the period from January 2002 to June 2011 at Songklanagarind Hospital, a tertiary care hospital in southern Thailand. Results: The prevalence of melioidosis among admitted patients was 36.8 per 100 000 in patients. The median age was 49 years and they were predominantly male. The most common underlying disease was diabetes mellitus (47.01%). The majority of cases (50%) had localized infection. The rates of multifocal, bacteremic, and disseminated infections were 12.7%, 23.1%, and 14.2%, respectively. The lungs were the most common organ afflicted, resulting in infection (24.63%). Splenic abscess as well as liver abscess accounted for 20.90% and 19.40%, respectively. A total of one eighth of the patients had septic shock at presentation. The overall mortality rate was 8.96%. The factors influencing mortality were pneumonia, septic shock, a positive blood culture for Burkholderia pseudomallei, superimposing with nosocomial infection and inappropriate antibiotic administration. Conclusions: Melioidosis is not uncommon in southern Thailand. The mortality of patients with pneumonia, bacteremia and septic shock is relatively high. Appropriate antibiotics, initially, will improve outcomes.
    10  Effect of dimethyl fumarate on rats with chronic pancreatitis
    Wen-Xian Zhang Jian-Hong Zhao Fu-Min Ping Zhi-Jun Liu Jun-Xia Gu Xin-Qing Lu
    2016, 9(3):256-259. DOI: 10.1016/j.apjtm.2016.01.023
    [Abstract](39) [HTML](0) [PDF 646.83 K](222)
    Objective: To discuss the effect of dimethyl fumarate (DMF) on rats with L-arginine induced chronic pancreatitis (CP). Methods: Male Wistar rats were given DMF treatment (25 mg/kg) by
    11  Onco-microRNA miR-130b promoting cell growth in children APL by targeting PTEN
    Xiang-Cui Gong Yuan-Qin Xu Yan Jiang Hui Guan Hua-Lin Liu
    2016, 9(3):260-263. DOI: 10.1016/j.apjtm.2016.01.024
    [Abstract](91) [HTML](0) [PDF 660.04 K](241)
    Objective: To study the expression of microRNA-130b (miR-130b) in children acute promyelocytic leukemia (APL) and its role for regulating PTEN expression. Methods: A total of 50 children APL marrow tissues and 15 normal marrow tissues between January and December in 2012 were collected into our study. The expression of miR-130b in APL and normal marrow tissues were detected by quantitative real-time polymerase chain reaction. MiR-130b inhibitor was transfected into HL-60 cells. Cell Counting Kit-8 assay and flow cytometry were used to measure cell proliferation and apoptosis, respectively. The expression of PTEN, a potential target of miR-130b, and its downstream genes, Bcl-2 and Bax, in transformed cells were detected by quantitative real-time polymerase chain reaction and western-blot. Results: The expression of miR-130b was significantly higher in children APL marrow tissues than in normal marrow tissues (P<0.05). Down-regulation of miR-130b could significantly suppress cell proliferation and induce apoptosis in HL-60 cells (P<0.05). PTEN expression was upregulated when miR-130b was knocking-down (P<0.05). As downstream genes of PTEN, the expression of Bcl-2 and Bax were regulated as well. Conclusions: MiR130b is overexpressed in children APL marrow tissues and associated with cell growth. MiR130b may promote children APL progression by inducing cell proliferation and inhibiting apoptosis.
    12  Effect of levocarnitine on TIMP-1, ICAM-1 expression of rats with coronary heart disease and its myocardial protection effect
    Shi-Xun Wang Lei Tan Jian Wang Jing-Quan Zhong
    2016, 9(3):264-268. DOI: 10.1016/j.apjtm.2016.01.025
    [Abstract](48) [HTML](0) [PDF 675.65 K](249)
    Objective: To study the effect of levocarnitine (L-CN) on tissue inhibitor of metalloproteinase-1 (TIMP-1) and intercellular adhesion molecule-1 (ICAM-1) expression of rats with coronary heart disease and evaluate the protective effect of L-CN on myocardial cells. Methods: High-fat diet feeding and intraperitoneal injection of pituitrin was performed on rats in model group and CHD Model of rats was built. Rats with successful model-building were selected and divided into L-CN group and Ctrl group randomly. Rats in L-CN group were given L-CN treatment, with intraperitoneal injection of 200 mgkg-1d-1 and successive administration for 3 d. Rats in Ctrl group were given equal volumes of normal saline. Blood was collected from carotid artery at different time and expression quantity of creatine kinase-MB (CK-MB) and Troponin Ⅰ (TnⅠ) in serum was detected. Rats in each group were put to death and were separated to obtain the myocardial tissue. Real-time PCR and Western Blotting hybridization were performed to detect the TIMP-1, ICAM-1 expression in myocardial tissue in each group. Statistical analysis was employed to explore the expression changes of TIMP-1 and ICAM-1, and ELISA test was used to analyse the expression changes of myocardial necrosis marker- CK-MB and TnⅠ to learn the effect of L-CN and its myocardial protective effect. Results: The total cholesterol, triglyceride and blood glucose levels of rats in model group were significantly higher than that in control group, which indicated that due to high-fat diet feeding, blood lipid of rats in model group was obviously higher than that in control group. In myocardial tissue of rats in model group, TIMP-1 level significantly reduced and ICAM-1 level significantly increased (P<0.01). In model group, after L-CN treatment, TIMP-1 level had double increase, while ICAM-1 level had 43% of decrease in L-CN group compared with Ctrl group. After L-CN intervention treatment, CK-MB and TnⅠ content in L-CN group relatively reduced compared with Ctrl group. The difference among groups was obvious (P<0.01). Conclusions: L-CN could increase the TIMP-1 expression level and inhibit the ICAM-1 expression level. L-CN has a certain myocardial protective effect.
    13  Regulating effect of activated NF-kB on edema induced by traumatic brain injury of rats
    Zi-Ran Wang Yu-Xin Li Hong-Yan Lei Dai-Qun Yang Li-Quan Wang Ming-Yu Luo
    2016, 9(3):269-272. DOI: 10.1016/j.apjtm.2016.01.027
    [Abstract](59) [HTML](0) [PDF 684.86 K](308)
    Objective: To observe the effect of nuclear transcription factor-κB (NF-κB) on cerebral edema in rats with traumatic brain injury (TBI). Methods: Male SD rats with fluid percussion injury (FPI) were selected. After separation and culture, rats’ astrocytes all suffered FPI. The expression of NF-κB and the water content were detected at the animal and cellular levels, while the activity of NOX was evaluated at the cellular level. Results: According to the results, the positive expression of NF-κB and expression of mRNA were significantly increased and the water content was increased for rats after TBI, while NF-κB inhibitor BAY11-7082 could significantly reduce the effect of TBI. 1 and 3h after FPI of astrocytes, the activation of NFκB was increased and BAY 11-7082 could significantly improve the injury-induced swelling of astrocytes. After the injury of astrocytes, the activity of NOX was also increased, while BAY 11-7082 could reduce the activity of NOX. Conclusions: The results show that the activation of NF-kB in astrocytes is a key factor in the process of cerebral edema after TBI of rats.
    14  Mechanism of all-transretinoic acid increasing retinoblastoma sensitivity to vincristine
    Yan Jiang Lin Zhang
    2016, 9(3):273-277. DOI: 10.1016/j.apjtm.2016.01.028
    [Abstract](48) [HTML](0) [PDF 652.86 K](208)
    Objective: To explore the mechanism of all-transretinoic acid (ATRA) increasing retinoblastoma (RB) sensitivity to vincristine, and the inhibiting effect of vincristine combined with ATRA treatment on the SO-RB50 cell proliferation. Methods: SO-RB50 cells were cultivated by routine culture method. Different concentrations of vincristine or ATRA were added into culture solution. After 48 h, cell counting kit-8 was used to detect the median inhibitory concentration (IC50) of vincristine combined with ATRT treatment to SO-RB50 cells. SO-RB50 cells were divided into drug combination group, vincristine group, ATRA group and control group. Different drugs were added into the culture solution respectively for cell culture based on the IC50 value. Cell counting kit-8 was used to detect the cell proliferation every 24-h cultivation. After continuous determination for 6 d, data was processed to draw the cell growth curve. After drug use for 72 h, flow cytometry was used to detect the proportion of different cell cycles of SO-RB50 cells in each group. After drug use for 48 h, annexin V/ propidium iodide method was used to detect the SO-RB50 cell apoptosis in each group. Results: The IC50 value of vincristine treatment on the SO-RB50 cells was 0.11 μmol/L, and ATRT was 12.84 μmol/L. The cell growth curve in control group rose gradually along with the extended culture time, but after vincristine and ATRA treatment, the cell growth curve was smooth and steady. The cell increment was the least in drug combination group and its cell growth curve was the smoothest. There was significant difference in A450 48 h and 72 h after treatment (Fgrouping=77.316, P<0.001; Ftime=86.985, P<0.001). Compared with control group, A450 value in drug combination group, vincristine group, ATRA group was significantly lower (P<0.001). Compared with control group, the G2/M phase cell proportion in vincristine group was significantly increased, while the G0/G1 phase cell proportion was significantly decreased; the G0/G1 phase cell proportion in ATRA group was significantly increased, while the S phase cell proportion was significantly decreased (FG0/G1=85.878, Fs=56.455, FG2/M=85.878, P<0.001). After 48 h, there was significant difference in SO-RB50 cell apoptosis rate among groups (F=11.312, P<0.05). The apoptosis rate in drug combination group was significantly higher than that of other groups (P<0.001). Conclusions: ATRA can increase the sensitivity of SO-RB50 cells to vincristine. Vincristine combined with ATRA treatment can significantly increase the inhibiting effect on SO-RB50 cells, which may be related with promoting cell apoptosis and involving in cell cycle control.
    15  Protective effect of apoptosis signal-regulating kinase1 inhibitor against mice liver injury
    Ping He Bo Zeng Xiao-Li Zhang Dian-Liang Fang Xia-Qia Zhou Ke-Qiang Wan Wen-Guang Tian
    2016, 9(3):278-282. DOI: 10.1016/j.apjtm.2016.01.029
    [Abstract](64) [HTML](0) [PDF 651.25 K](232)
    Objective: To explore the protective effect and its molecular mechanism of apoptosis signal-regulating kinase1 (ASK1) inhibitor (GS-459679) on acetaminophen-induced liver injury in mice. Methods: The model of liver injury was established by administration of acetaminophen (APAP) (300 mg/kg, i.p.) on C57BL/6 mice. Forty-eight male C57BL/6 mice were randomly divided into four groups, consisting of control group, GS group (GS-459679, 30 mg/kg, i.p.), APAP-induced group, and GS combined with APAP-induced group. For GS combined with APAP-induced group, mice were treated with GS 30 min prior to administration of APAP. After mice were euthanized at 6 h or 12 h, respectively, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed, and mRNA levels of /i>TNF-α, IL-6 and IL-1β were tested. The activity of glutathione (GSH), oxidized GSH (GSSG) and malondialdehyde were quantified. In addition, ASK1,P-ASK1, JNK and P-JNK protein levels were tested in all groups. Results: The ASK1 and P-ASK1 levels were up-regulated in APAP-induced group. Compared to the control group, serum levels of ALT and AST, and mRNA levels of TNF-α, IL-6 and IL-1β were increased in APAP-induced group. Meanwhile, the levels of MAD and GSSG, and the ratio of GSSG/GSH were higher and the JNK was activatedin APAP-induced group compared with that in control group. However, compared to APAP-induced group, GS combined with APAP-induced group displayed a decrease of protein expression levels of ASK1, P-ASK1 and P-JNK, a reduction of serum levels of ALT and AST, a decrease in TNF-α, IL-6 and IL-1β mRNA levels, and a low ration of GSSG/GSH. Conclusions: GS-459679 treatment effectively down-regulates ASK1 and P-ASK1 expression. Addition of GS-459679 decreases the generation of liver metabolites and inflammatory factors, reduces oxidative stress reaction, inhibits JNK activation, and then protects the responsiveness to APAP-induced liver injury.
    16  Effect of TRPV1 combined with lidocaine on cell state and apoptosis of U87-MG glioma cell lines
    Jun Lu You-Ting Ju Chang Li Fu-Zhou Hua Guo-Hai Xu Yan-Hui Hu
    2016, 9(3):283-287. DOI: 10.1016/j.apjtm.2016.01.030
    [Abstract](44) [HTML](0) [PDF 758.43 K](260)
    Objective: To study the effects of Transient receptor potential cation channel, subfamily V, member 1 (TRPV1) combined with lidocaine on status and apoptosis of U87-MG glioma cell line, and explore whether local anesthetic produces neurotoxicity by TRPV1. Methods: U87MG cells were divided into control group, gene silencing group, empty vector group and TRPV gene up-regulation group. For cells in each group, flow cytometry was employed to detect the intracellular calcium ion concentration and mitochondrial membrane potential at different time point from cellular perspective. Cell apoptosis of U87-MG was assayed by flow cytometry and MTT from a holistic perspective. Results: Calcium ion concentration increased along with time. The concentration in TRPV1 gene up-regulation group was significantly higher than those in other groups at each time point (P<0.05). After adding lidocaine, mitochondrial membrane potential in U87-MG significantly increased (P<0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P<0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P<0.05). Flow cytometry result and MTT result both showed that cell apoptosis in each group significantly increased after lidocaine was added (P<0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P<0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P<0.05). Moreover, apoptosis was more severe along with the increasing concentration of lidocaine (P<0.05). Conclusions: In this study, it was proved that lidocaine could dose-dependently induce the increase of intracellular calcium ion concentration, mitochondrial membrane potential and apoptosis in U87-MG glioma cell line. The up-regulation of TRPV1 enhanced cytotoxicity of lidocaine, which revealed the correlations between them. Lidocaine might have increased intracellular calcium ion concentration by activating TRPV1 gene and induced apoptosis of U87GM glioma cell line by up-regulating mitochondrial membrane potential.
    17  Immune formulation-assisted conventional therapy on anti-infective effectness of multidrug-resistant Mycobacterium tuberculosis infection mice
    Xiu-Li Yuan Qiang Wen Ming-De Ni Li-Kun Wang
    2016, 9(3):288-292. DOI: 10.1016/j.apjtm.2016.01.031
    [Abstract](65) [HTML](0) [PDF 694.54 K](268)
    Objective: To study the effect of immune formulation-assisted conventional therapy on anti-infective ability of multidrug-resistant Mycobacterium tuberculosis infection mice. Methods: BALB/c mice were used as experimental animals, multidrug-resistant Mycobacterium tuberculosis infection models were built, randomly divided into model group, moxifloxacin group, thymopentin group and combined treatment group and given corresponding drug intervention, and then colony numbers in the spleen and lung, T lymphocyte subset contents and programmed death-1 (PD-1) expression levels in peripheral blood were detected. Results: Colony numbers in lung and spleen of moxifloxacin group and thymopentin group were significantly lower than those of model group and colony numbers in lung and spleen of combined treatment group were significantly lower than those of moxifloxacin group and thymopentin group; contents of CD3+CD4+T cells, Th1 and Th17 in peripheral blood of moxifloxacin group and thymopentin group were higher than those of model group, and contents of CD3+CD8+T cells, Th2 and Treg were lower than those of model group; contents of CD3+CD4+T cells, Th1 and Th17 in peripheral blood of combined treatment group were higher than those of moxifloxacin group and thymopentin group, and contents of CD3+CD8+T cells, Th2 and Treg were lower than those of moxifloxacin group and thymopentin group; PD-1 expression levels on T lymphocyte, B lymphocyte and monocyte surface in peripheral blood of moxifloxacin group and thymopentin group were lower than those of model group, and PD-1 expression levels on T lymphocyte, B lymphocyte and monocyte surface in peripheral blood of combined treatment group were lower than those of moxifloxacin group and thymopentin group. Conclusions: Immune formulation thymopentin can enhance the anti-infective ability of multidrug-resistant Mycobacterium tuberculosis infection mice, decrease bacterial load in lung and spleen, and enhance immune function.
    18  Effect of miR-467b on atherosclerosis of rats
    Xiao-Mei Guan Yong-Xin Li Hai Xin Jun Li Zong-Gang Zhao Yue-Wei Wang Hao-Fu Wang
    2016, 9(3):293-296. DOI: 10.1016/j.apjtm.2016.01.026
    [Abstract](57) [HTML](0) [PDF 662.55 K](264)
    Objective: To observe the effect of miR-467b on the atherosclerosis (AS) of rats with apolipoprotein E (ApoE) gene knockout (ApoE-/-). Methods: ApoE-/-rats were fed with high fat and high cholesterol diet and were randomly divided into group A, group B and group C, with 10 rats in each group. Group A: rats were injected with ApoE agonist through the caudal vein; Group B: rats were injected with ApoE antagonist through the caudal vein; Group C: as negative control group. Enzyme oxidation method was used to detect the blood lipid levels of rats. Western blotting method was used to detect the aortic lipoprotein lipase (LPL) expression levels of rats. HE staining and oil red o staining were performed to observe the AS lesions and lipid accumulation state. Results: Compared with Group C, blood lipid level, aortic intima and aortic sinus lipid accumulation area ratio, aortic sinus lesion area and LPL expression level in Group A significantly reduced; while blood lipid level, aortic intima and aortic sinus lipid accumulation area ratio, aortic sinus lesion area, and LPL expression level in Group B significantly increased, with the statistical difference (P<0.05). Conclusions: miR-467b can alleviate the AS lesions of ApoE-/-rats, and its inhibiting effect on AS may be related to LPL expression.
    19  Preinduced intestinal HSP70 improves visceral hypersensitivity and abnormal intestinal motility in PI-IBS mouse model
    Cheng Lan Xiao-Ning Sun Xu-Chun Zhou Bo Yang Bai-Li Huang Tao-Zhi Deng Zhou-Tao He Xiang-Yang Han
    2016, 9(3):297-300. DOI: 10.1016/j.apjtm.2016.01.022
    [Abstract](45) [HTML](0) [PDF 701.70 K](232)
    Objective: To investigate the impact of the preinduced intestinal heat shock protein 70 (HSP70) on the visceral hypersensitivity and abnormal intestinal motility in a post-infectious irritable bowel syndrome (PI-IBS) mouse model. Methods: Eighty-four female C57BL/6 mice were randomly assigned to four groups: control group (n=21) and induction+PI-IBS group (n=21), PI-IBS group (n=21) and induction group (n=21). The mice in PI-IBS group were infected in vivo with trichinella spiralis by oral administration. The visceral hypersensitivity and intestinal motility were evaluated respectively with abdominal withdrawal reflex and colon transportation test. The intestinal HSP70 protein?? and mRNA level was measured by Western blot and real-time PCR. Meanwhile, the intestinal proinflammatory cytokines IL-10 and TNF-α level was detected by ELISA. Results: Compared with their counterparts in PI-IBS group, the animals in the Induction+PI-IBS group show significantly increased intestinal level of HSP70 and obviously ameliorative clinical figures, including abdominal withdrawal reflex score, intestine transportation time and Bristol scores (P<0.05). Meanwhile, the intestinal post-inflammatory cytokines remarkably changed, including increased IL-10 level and decreased TNF-α level (P<0.05). Conclusions: Intestinal HSP70 may play a potential protective role through improving the imbalance between the intestinal post-inflammatory and anti-inflammatory cytokines in PI-IBS.

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