Inhibition of Autophagy Promotes the Elimination of Liver Cancer Stem Cells by CD133 Aptamer-Targeted Delivery of Doxorubicin.

Doxorubicin is the most frequently used chemotherapeutic agent for the treatment of hepatocellular carcinoma. However, one major obstacle to the effective management of liver cancer is the drug resistance derived from the cancer stem cells. Herein, we employed a CD133 aptamer for targeted delivery of doxorubicin into liver cancer stem cells to overcome chemoresistance. Furthermore, we explored the efficacy of autophagy inhibition to sensitize liver cancer stem cells to the treatment of CD133 aptamer-doxorubicin conjugates based on the previous observation that doxorubicin contributes to the survival of liver cancer stem cells by activating autophagy. The kinetics and thermodynamics of aptamer-doxorubicin binding, autophagy induction, cell apoptosis, and self-renewal of liver cancer stem cells were studied using isothermal titration calorimetry, Western blot analysis, annexin V assay, and tumorsphere formation assay. The aptamer-cell binding andintracellular accumulation of doxorubicin were quantified via flow cytometry. CD133 aptamer-guided delivery of doxorubicin resulted in a higher doxorubicin concentration in the liver cancer stem cells. The combinatorial treatment strategy of CD133 aptamer-doxorubicin conjugates and an autophagy inhibitor led to an over 10-fold higher elimination of liver cancer stem cells than that of free doxorubicin in vitro. Future exploration of cancer stem cell-targeted delivery of doxorubicin in conjunction with autophagy inhibition in vivo may well lead to improved outcomes in the treatment of hepatocellular carcinoma.

Neuropeptide Y modifies a part of diencephalic catecholamine but not indolamine metabolism in chicks depending on feeding status.

The role of the monoaminergic system in the feeding behavior of neonatal chicks has been reported, but the functional relationship between the metabolism of monoamines and appetite-related neuropeptides is still unclear. This study aimed to investigate the changes in catecholamine and indolamine metabolism in response to the central action of neuropeptide Y (NPY) in different feeding statuses and the underlying mechanisms. In Experiment 1, the diencephalic concentrations of amino acids and monoamines following the intracerebroventricular (ICV) injection of NPY (375 pmol/10 µl/chick), saline solution under ad libitum, and fasting conditions for 30 min were determined. Central NPY significantly decreased L-tyrosine concentration, the precursor of catecholamines under feeding condition, but not under fasting condition. Central NPY significantly increased dopamine metabolites, including 3,4-dihydroxyphenylacetic acid and homovanillic acid (HVA). The concentration of 3-methoxy-4-hydroxyphenylglycol was significantly reduced under feeding condition, but did not change under fasting condition by NPY. However, no effects of NPY on indolamine metabolism were found in either feeding status. Therefore, the mechanism of action of catecholamines with central NPY under feeding condition was elucidated in Experiment 2. Central NPY significantly attenuated diencephalic gene expression of catecholaminergic synthetic enzymes, such as tyrosine hydroxylase, L-aromatic amino acid decarboxylase, and GTP cyclohydrolase I after 30 min of feeding. In Experiment 3, co-injection of α-methyl-L-tyrosine, an inhibitor of tyrosine hydroxylase with NPY, moderately attenuated the orexigenic effect of NPY, accompanied by a significant positive correlation between food intake and HVA levels. In Experiment 4, there was a significant interaction between NPY and clorgyline, an inhibitor of monoamine oxidase A with ICV co-injection which implies that co-existence of NPY and clorgyline enhances the orexigenic effect of NPY. In conclusion, central NPY modifies a part of catecholamine metabolism, which is illustrated by the involvement of dopamine transmission and metabolism under feeding but not fasting conditions.

Central GABAA receptor mediates taurine-induced hypothermia and possibly reduces food intake in thermo-neutral chicks and regulates plasma metabolites in heat-exposed chicks.

The aim of this study was to examine the central action of taurine on body temperature and food intake in neonatal chicks under control thermoneutral temperature (CT) and high ambient temperature (HT). Intracerebroventricular injection of taurine caused dose-dependent hypothermia and reduced food intake under CT. The mRNA expression of the GABAA receptors, GABAAR-α1 and GABAAR-γ, but not that of GABABR, significantly decreased in the diencephalon after central injection of taurine. Subsequently, we found that picrotoxin, a GABAAR antagonist, attenuated taurine-induced hypothermia. Central taurine significantly decreased the brain concentrations of 3-methoxy-4-hydroxyphenylglycol, a major metabolite of norepinephrine; however, the concentrations of serotonin, dopamine, and the epinephrine metabolites, 3,4-hydroxyindoleacetic acid and homovanillic acid, were unchanged. Although hypothermia was not observed under HT after central injection of taurine, plasma glucose and uric acid levels were higher, and plasma sodium and calcium levels were lower, than those in chicks under CT. In conclusion, brain taurine may play a role in regulating body temperature and food intake in chicks through GABAAR. The changes in plasma metabolites under heat stress suggest that brain taurine may play an important role in maintaining homeostasis in chicks.

The inhibition of ABCB1/MDR1 or ABCG2/BCRP enables doxorubicin to eliminate liver cancer stem cells.

Two ATP-binding cassette transporters, ABCB1/MDR1 and ABCG2/BCRP, are considered the most critical determinants for chemoresistance in hepatocellular carcinoma. However, their roles in the chemoresistance in liver cancer stem cells remain elusive. Here we explored the role of inhibition of MDR1 or ABCG2 in sensitizing liver cancer stem cells to doxorubicin, the most frequently used chemotherapeutic agent in treating liver cancer. We show that the inhibition of MDR1 or ABCG2 in Huh7 and PLC/PRF/5 cells using either pharmacological inhibitors or RNAi resulted in the elevated level of intracellular concentration of doxorubicin and the accompanied increased apoptosis as determined by confocal microscopy, high-performance liquid chromatography, flow cytometry, and annexin V assay. Notably, the inhibition of MDR1 or ABCG2 led to the reversal of the chemoresistance, as evident from the enhanced death of the chemoresistant liver cancer stem cells in tumorsphere-forming assays. Thus, the elevation of effective intracellular concentration of doxorubicin via the inhibition of MDR1 or ABCG2 represents a promising future strategy that transforms doxorubicin from a traditional chemotherapy agent into a robust killer of liver cancer stem cells for patients undergoing transarterial chemoembolization.

Icariin reduces bone loss in a Rankl-induced transgenic medaka (Oryzias latipes) model for osteoporosis.

Given the limitations and side effects of many synthetic drugs, natural products are an important alternative source for drugs and medications for many diseases. Icariin (ICA), one of the main flavonoids from plants of the Epimedium genus, has been shown to ameliorate osteoporosis and improve bone health in preclinical studies. Those studies have used different in vivo models, mostly rodents, but the underlying mechanisms remain unclear. The present study shows, for the first time, that ICA reduces bone damage in a Rankl-induced medaka fish (Oryzias latipes), a non-rodent osteoporosis model. Live imaging was previously performed in this model to characterize antiresorptive and bone-anabolic properties of drugs. Here, a new quantification method (IM ) was established based on the length of mineralized neural arches to quantify levels of bone mineralization damage and protection in early post-embryonic fish. This method was validated by quantification of three levels of bone damage in three independent Rankl fish lines, and by the determination of different degrees of severity of osteoporosis-like phenotypes in one Rankl line exposed to variable Rankl induction schemes. IM was also used to quantify the efficacy of alendronate and etidronate, two common anti-osteoporotic bisphosphonates, and revealed comparable bone protective effects for ICA and alendronate in this fish osteoporosis model. This study's data support the value of the medaka fish model for bone research and establish a method to screen for novel osteoprotective compounds.

Oral Administration of Watermelon Rind Extract to Induce Hypothermia in Chicks.

Oral administration of l-citrulline (l-Cit) caused hypothermia, but l-Cit is not recommended in poultry diets in Japan. Watermelon is a natural source of l-Cit. The objective of this study is to examine the effect of watermelon waste, i.e., watermelon rind (WR) on the body temperature and plasma free amino acids of chicks. In Experiment 1, 14-day-old chicks were subjected to acute oral administration of WR extract (WRE) (2 ml) under control thermoneutral temperature (CT). In Experiment 2, 15-day-old chicks were orally administered 1.6 ml of either WRE, lowdose l-Cit (7.5 mmol/10 ml), or high-dose l-Cit (15 mmol/10 ml) under CT. In both experiments, rectal temperature (RT) and plasma free amino acids were analyzed. In Experiment 3, after dual oral administration of (1.6 ml) WRE or l-Cit (15 mmol/10 ml), 15-day-old chicks were exposed to high ambient temperature (HT; 35±1°C, 2 h) to monitor changes in RT. Acute oral administration of WRE significantly reduced RT under CT. The degree of RT reduction by WRE was similar to that by high l-Cit. Moreover, RT was significantly low at HT owing to the oral administration of WRE. However, the reduced RT was difficult to explain by the content of Cit in WRE alone. In conclusion, WRE could be used as a dietary ingredient to reduce body temperature for imparting thermotolerance in chicks.

An acute increase in water temperature can increase free amino acid concentrations in the blood, brain, liver, and muscle in goldfish (Carassius auratus).

Water temperature directly affects the body temperature in fish, so increasing water temperatures in oceans and rivers will lead to increases in fish body temperatures. Whilst a range of responses of fish to increases in water temperature have been measured, amino acid metabolism in a fish under high water temperature (HT) conditions has not been investigated. The aim of this study was to determine the effects of an acute increase in water temperature on oxygen consumption, plasma cortisol concentrations, and free amino acid concentrations in plasma and several tissues in goldfish (Carassius auratus). Oxygen consumption and plasma cortisol concentrations were increased in goldfish exposed to HT (30 ± 1 °C) for 200 min compared with goldfish at a control water temperature (CT 17 ± 1 °C). Oxygen consumption and plasma cortisol concentrations in both groups of fish combined were positively correlated. When goldfish were exposed to HT for 300 min oxygen consumption and plasma concentrations of 15 free amino acids were increased compared with goldish at CT. Concentrations of several free amino acids were increased to varying extents in the brain, liver, and muscle tissues. In conclusion, an acute increase in water temperature affected amino acid metabolism differently in the brain, liver, and muscle tissues. Goldfish will be a useful species for further studies of the possible roles of various amino acids in the brain, muscle, and liver during acute increases in water temperature in fish.

Alcohol Consumption and Binge Drinking Among Adult Population: Evidence From the CHILILAB Health and Demographic Surveillance System in Vietnam.

CONTEXT: Alcohol contributes to severe social and health problems and is a major risk factor for noncommunicable diseases in Vietnam. Over the years, there has been an increase in consumption per capita as well as a rapid expansion of commercially prepared alcohol. OBJECTIVE: To describe the prevalence of alcohol consumption and binge drinking in a random sample of people 15 years of age and older living in Chi Linh who were also a part of the Chi Linh Health and Demographic Surveillance System (CHILILAB HDSS) and to determine the association between alcohol use and sociodemographic characteristics. DESIGN: Data on alcohol consumption of 5438 people 15 years of age and older were extracted from the CHILILAB HDSS information collected in 2016. Descriptive statistics and multiple logistic regression were utilized to assess the association between current drinkers and binge drinkers with socioeconomic groups. RESULTS: The overall prevalence of alcohol use 1 month prior to interview was 41.1%, which is composed of 75.1% males and 17.3% females. Among the 41% of alcohol drinkers, 31.7% reported binge drinking over the last 30 days. The proportion of binge drinking was also found to be higher among males than among females. The association between current drinkers, binge drinkers and gender, area of residence, education, and family income level was statistically significant. CONCLUSIONS: This study confirms that alcohol use among current and binge drinkers is common among males in Vietnam and that it is also a rising issue among females. Alcohol use is also associated with sociodemographic factors and income level. The results of this study provide evidence of harmful alcohol use among the Vietnamese population, which could help policy makers further advocate for the approval of the Vietnamese alcohol harm reduction law in the coming years. The results of this study reaffirm the need for public health strategies, including the formulation of laws and policies to reduce the harmful effects of alcohol consumption in Vietnam.

Injury Mortality in Vietnam: Patterns and Trends, 2005-2013.

CONTEXT: Injury is a growing public health problem worldwide. Deaths due to injuries account for 10% of the world's mortality. More than 90% of the world's injury-related deaths occur in low- and middle-income countries such as Vietnam. The public health burden has been recognized by government; however, there is a need to study and better understand the epidemiology and trends for injuries and injury-related mortality to be able to better address this burden. OBJECTIVES: In this article, we analyze the patterns and trends in injury mortality from 2005 to 2013 in Vietnam. DESIGN: All registered deaths from injuries from January 2005 to December 2013 were extracted from the Ministry of Health death recording system (A6). Mortality rates per 100 000 population per year were calculated. A linear regression model was used to estimate the injury mortality trends. RESULTS: In the 9-year period, 313 101 deaths due to injuries were recorded in the A6 system; this accounts for about 10% of all deaths. The leading causes of injury-related death were road traffic injuries among the entire population, while drowning was the leading cause of death among children. Other unintentional injuries including occupational, fall, poisoning, and exposure to electric current also accounted for a substantial proportion of the burden of injury. There is a significant reduction trend found in drowning among children 0 to 18 years of age and significant increasing trends in intentional injury causes. CONCLUSIONS: While injuries have been recognized as a public health problem in Vietnam in the last decade, as seen by our analysis, there is a need for concerted action to reduce their burden. Of particular concern is the increasing prominence of intentional injuries. Improved data systems, increasing the awareness in the community, and making appropriate policies and implementing them, as well as implementing effective, evidence-based interventions are all key to decreasing this burden. This is an important study describing injury mortality in Vietnam for the period 2005-2013. The results of the study show that injury death remains an important public health issue that needs more attention from government and relevant agencies.

Cytotoxic Clerodane Diterpenoids from the Leaves of Casearia grewiifolia.

Two new clerodane diterpenoids (1 and 2) and the known compound caseanigrescen D (3) were isolated from the leaves of Casearia grewiifolia by bioassay-guided fractionation. Their structures were determined by analyses of MS and 2D NMR data. The absolute configurations of 1 and 3 were established by analysis of X-ray diffraction data. Compounds 1-3 were evaluated for their cytotoxicity against four cancer cell lines, KB (mouth epidermal carcinoma cells), HepG-2 (human liver hepatocellular carcinoma cells), LU-1 (human lung adenocarcinoma cells), and MCF-7 (human breast cancer cells). Caseagrewifolin B (2) had inhibitory activity against KB and HepG-2 cell lines with IC50 values of 6.2 to 7.0 µM, respectively. When tested against the normal cells (NIH/3T3), caseagrewifolin B (2) exhibited a significant selective inhibition against cancer cells in comparison with the normal cells. Caseanigrescen D (3) was cytotoxic against four cancer cell lines; however it had no selective inhibition compared with normal cells.

Factors associated with suboptimal adherence to antiretroviral therapy in Viet Nam: a cross-sectional study using audio computer-assisted self-interview (ACASI).

BACKGROUND: Optimal adherence to antiretroviral therapy (ART) is necessary for people living with HIV/AIDS (PLHIV). There have been relatively few systematic analyses of factors that promote or inhibit adherence to antiretroviral therapy among PLHIV in Asia. This study assessed ART adherence and examined factors associated with suboptimal adherence in northern Viet Nam. METHODS: Data from 615 PLHIV on ART in two urban and three rural outpatient clinics were collected by medical record extraction and from patient interviews using audio computer-assisted self-interview (ACASI). RESULTS: The prevalence of suboptimal adherence was estimated to be 24.9% via a visual analogue scale (VAS) of past-month dose-missing and 29.1% using a modified Adult AIDS Clinical Trial Group scale for on-time dose-taking in the past 4 days. Factors significantly associated with the more conservative VAS score were: depression (p < 0.001), side-effect experiences (p < 0.001), heavy alcohol use (p = 0.001), chance health locus of control (p = 0.003), low perceived quality of information from care providers (p = 0.04) and low social connectedness (p = 0.03). Illicit drug use alone was not significantly associated with suboptimal adherence, but interacted with heavy alcohol use to reduce adherence (p < 0.001). CONCLUSIONS: This is the largest survey of ART adherence yet reported from Asia and the first in a developing country to use the ACASI method in this context. The evidence strongly indicates that ART services in Viet Nam should include screening and treatment for depression, linkage with alcohol and/or drug dependence treatment, and counselling to address the belief that chance or luck determines health outcomes.

Child injury prevention in Vietnam: achievements and challenges.

As with other South Asian countries, injury is becoming a leading cause of death and morbidity among children in Vietnam. In response to the increasing burden of child injury, government and non-government agencies in Vietnam have combined efforts during the last decade to develop and implement various child injury prevention strategies and programmes. This article provides, through a review of relevant documents and interviews with major stakeholders, an overview of these efforts and highlights major challenges to child injury prevention in the country. The findings point to notable achievements in terms of increasing awareness of injury facing children at all levels in the community and developing a sound injury prevention policy framework in a relatively short period of time. However, much needs to be done to implement necessary environmental and legislative changes, strengthen child injury surveillance and injury prevention research; and to improve access to health services. The insight into the experience of Vietnam could benefit other low- and middle-income countries with a high burden of child injury.

Antituberculosis cycloartane triterpenoids from Radermachera boniana.

Three new triterpenoids, bonianic acids A (1) and B (2) and 3-O-acetyluncaric acid (3), were isolated from the leaves and twigs of Radermachera boniana, together with six known compounds, ursolic acid (4), oleanolic acid (5), 3-epi-oleanolic acid (6), 3α-O-acetyl-α-boswellic acid (7), ergosterol peroxide (8), and ß-sitostenone (9). Ergosterol peroxide (8) and bonianic acids A (1) and B (2) exhibited significant activity against Mycobacterium tuberculosis H37Rv strain.

Occupational injuries as reported in the Vietnam Multi-Center Injury Survey.

Hundreds of deaths and thousands of serious injuries occur at workplace every year in Vietnam, causing losses in life, labor productivity and burden to the society. However, these injuries are poorly reported and little effort has been made to prevent them. Using data from the Vietnam Multi-Center Injury Survey in 2001, this study sought to provide better data on the extent and nature of occupational injuries. The overall rate of occupational injuries was 26.01/1000 among people aged 18 years and older, with higher rates in rural than urban areas. Handicraft workers had the highest rate of injury (46.4/1000) followed by farmer and individuals working privately. These rates are all consistently higher than the rates from official reports. This study highlights the need for strengthening current occupational injury surveillance system, to provide better understanding of the problem for setting priority and creating evidence based prevention strategies.