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  1. Fulltext Promotion of HepG2 cell apoptosis by flower of Allium atroviolaceum and the mechanism of action
    Khazaei S, Abdul Hamid R, Mohd Esa N, Ramachandran V, Aalam GT, Etemad A, et al.
    BMC Complement Altern Med, 2017 Feb 10;17(1):104.
    PMID: 28187719 DOI: 10.1186/s12906-017-1594-6
    BACKGROUND: Liver cancer is a high incidence and fatal disease, the fifth most frequent cancer worldwide that is usually diagnosed at an advanced stage. The number of deaths from liver cancer has not declined even following various therapies. Plant secondary metabolites and their semi-synthetic derivatives play a principal role in anti-cancer drug therapy, since they are effective in the treatment of specific characteristics while also reducing side effects. Allium atroviolaceum, a plant of the genus Allium has been used in folk medicine to protect against several diseases. However, cytotoxicity and the anti-proliferative effect of Allium atroviolaceum remain unclear. This work aims to investigate the anticancer properties of Allium atroviolaceum and the mechanism of action.

    METHODS: To evaluate the in vitro cytotoxicity of flower of Allium atroviolaceum, methanol extract at a dose range from 100 to 3.12 μg/ml was assessed against the HepG2 hepatocarcinoma cell line, and also on normal 3T3 cells, by monitoring proliferation using the MTT assay method. A microscopy study was undertaken to observe morphological changes of HepG2 cells after treatment and cell cycle arrest and apoptosis were studied using flow cytometry. The apoptosis mechanism of action was assessed by the level of caspase-3 activity and expression of apoptosis related genes, Bcl-2, Cdk1 and p53. The combination effect of the methanolic extract with doxorubicin was also investigated by determination of a combination index.

    RESULTS: The results demonstrated growth inhibition of cells in both dose- and time-dependent manners, while no cytotoxic effect on normal cell 3T3 was found. The results revealed the occurrence of apoptosis, illustrated by sub-G0 cell cycle arrest, the change in morphological feature and annexin-V and propidium iodide staining, which is correlated with Bcl-2 downregulation and caspase-3 activity, but p53-independent. In addition, a combination of Allium atroviolaceum and doxorubicin led to a significant synergistic effect.

    CONCLUSION: These findings suggest that Allium atroviolaceum flower extract has potential as a potent cytotoxic agent against HepG2 cell lines, as it has commendable anti-proliferative activities against human hepatocarcinoma and it can be considered as an effective adjuvant therapeutic agent after the clinical trials.

  2. Fulltext Analysis of selected glutathione S-transferase gene polymorphisms in Malaysian type 2 diabetes mellitus patients with and without cardiovascular disease
    Etemad A, Vasudevan R, Aziz AF, Yusof AK, Khazaei S, Fawzi N, et al.
    Genet. Mol. Res., 2016 Apr 07;15(2).
    PMID: 27173202 DOI: 10.4238/gmr.15025845
    Type 2 diabetes mellitus (T2DM) is believed to be associated with excessive production of reactive oxygen species. Glutathione S-transferase (GST) polymorphisms result in decreased or absent enzyme activity and altered oxidative stress, and have been associated with cardiovascular disease (CVD). The present study assessed the effect of GST polymorphisms on the risk of developing T2DM in individuals of Malaysian Malay ethnicity. A total of 287 subjects, consisting of 87 T2DM and 64 CVD/T2DM patients, as well as 136 healthy gender- and age-matched controls were genotyped for selected polymorphisms to evaluate associations with T2DM susceptibility. Genomic DNA was extracted using commercially available kits, and GSTM1, GSTT1, and α-globin sequences were amplified by multiplex polymerase chain reaction. Biochemical parameters were measured with a Hitachi autoanalyzer. The Fisher exact test, the chi-square statistic, and means ± standard deviations were calculated using the SPSS software. Overall, we observed no significant differences regarding genotype and allele frequencies between each group (P = 0.224 and 0.199, respectively). However, in the combined analysis of genotypes and blood measurements, fasting plasma glucose, HbA1c, and triglyceride levels, followed by age, body mass index, waist-hip ratio, systolic blood pressure, and history of T2DM significantly differed according to GST polymorphism (P ˂ 0.05). Genetically induced absence of the GSTT1 enzyme is an independent and powerful predictor of premature vascular morbidity and death in individuals with T2DM, and might be triggered by cigarette smoking's oxidative effects. These polymorphisms could be screened in other ethnicities within Malaysia to determine further possible risk factors.
  3. Fulltext Erratum to "Expression Profiling of Genes Related to Endothelial Cells Biology in Patients with Type 2 Diabetes and Patients with Prediabetes"
    Moradipoor S, Ismail P, Etemad A, Wan Sulaiman WA, Ahmadloo S, Khazaei S
    Biomed Res Int, 2017;2017:9764930.
    PMID: 28293642 DOI: 10.1155/2017/9764930
    [This corrects the article DOI: 10.1155/2016/1845638.].
  4. Flower extract of Allium atroviolaceum triggered apoptosis, activated caspase-3 and down-regulated antiapoptotic Bcl-2 gene in HeLa cancer cell line
    Khazaei S, Ramachandran V, Abdul Hamid R, Mohd Esa N, Etemad A, Moradipoor S, et al.
    Biomed Pharmacother, 2017 May;89:1216-1226.
    PMID: 28320088 DOI: 10.1016/j.biopha.2017.02.082
    Cervical cancer accounts for the second most frequent cancer and also third leading cause of cancer mortality (15%) among women worldwide. The major problems of chemotherapeutic treatment in cervical cancer are non-specific cytotoxicity and drug resistance. Plant-derived products, known as natural therapies, have been used for thousands of years in cancer treatment with a very low number of side effects. Allium atroviolaceum is a species in the genus Allium and Liliaceae family, which could prove to have beneficial effects on cancer treatment, although there is a lack of corresponding attention. The methanolic extract from the A.atroviolaceum flower displayed marked anticancer activity on HeLa human cervix carcinoma cells with much lower cytotoxic effects on normal cells (3T3). The A.atroviolaceum extract induced apoptosis, confirmed by cell cycle arrest at the sub-G0 (apoptosis) phase, characteristic morphological changes, evident DNA fragmentation, observed by fluorescent microscope, and early and late apoptosis detection by Annexin V. Furthermore, down-regulation of Bcl-2 and activation of caspase-9 and -3 strongly indicated that the mitochondrial pathway was involved in the apoptosis signal pathway. Moreover, combination of A.atroviolaceum extract with doxorubicin revealed a significant reduction of IC50and led to a synergistic effect. In summary, A.atroviolaceum displayed a significant anti-tumour effect through apoptosis induction in HeLa cells, suggesting that the A.atroviolaceum flower might have therapeutic potential against cervix carcinoma.
  5. Fulltext In vitro Antiproliferative and Apoptosis Inducing Effect of Allium atroviolaceum Bulb Extract on Breast, Cervical, and Liver Cancer Cells
    Khazaei S, Esa NM, Ramachandran V, Hamid RA, Pandurangan AK, Etemad A, et al.
    Front Pharmacol, 2017;8:5.
    PMID: 28197098 DOI: 10.3389/fphar.2017.00005
    Natural products are considered potent sources for novel drug discovery and development. The multiple therapeutic effects of natural compounds in traditional medicine motivate us to evaluate the cytotoxic activity of bulb of Allium atroviolaceum in MCF7 and MDA-MB-231, HeLa and HepG2 cell lines. The bulb methanol extract of A. atroviolaceum was found to be an active cell proliferation inhibitor at the time and dose dependent manner. Determination of DNA content by flow cytometry demonstrated S and G2/M phase arrest of MCF-7 cell, correlated to Cdk1 downregulation, S phase arrest in MDA-MB-231 which is p53 and Cdk1-dependent, sub-G0 cell cycle arrest in HeLa aligned with Cdk1 downregulation, G0/G1, S, G2/M phase arrest in HepG2 which is p53-dependent. Apoptosis as the mechanism of cell death was confirmed by morphology study, caspases activity assay, as well as apoptosis related gene expression, Bcl-2. Caspase-8, -9, and -3 activity with downregulation of Bcl-2 illustrated occurrence of both intrinsic and extrinsic pathways in MCF7, while caspase-3 and -8 activity revealed extrinsic pathway of apoptosis, although Bcl-2 downregulated. In HeLa cells, the activity of caspase-9 and -3 and downregulation of Bcl-2 shows intrinsic pathway or mitochondrial pathway, whereas HepG2 shows caspase independent apoptosis. Further, the combination of the extract with tamoxifen against MCF7 and MDA-MB-231 and combination with doxorubicin against HeLa and HeG2 demonstrated synergistic effect in most concentrations, suggests that the bulb of A. atroviolaceum may be useful for the treatment of cancer lonely or in combination with other drugs.
  6. Fulltext A full pipeline of diagnosis and prognosis the risk of chronic diseases using deep learning and Shapley values: The Ravansar county anthropometric cohort study
    Jafari H, Shohaimi S, Salari N, Kiaei AA, Najafi F, Khazaei S, et al.
    PLoS One, 2022;17(1):e0262701.
    PMID: 35051240 DOI: 10.1371/journal.pone.0262701
    Anthropometry is a Greek word that consists of the two words "Anthropo" meaning human species and "metery" meaning measurement. It is a science that deals with the size of the body including the dimensions of different parts, the field of motion and the strength of the muscles of the body. Specific individual dimensions such as heights, widths, depths, distances, environments and curvatures are usually measured. In this article, we investigate the anthropometric characteristics of patients with chronic diseases (diabetes, hypertension, cardiovascular disease, heart attacks and strokes) and find the factors affecting these diseases and the extent of the impact of each to make the necessary planning. We have focused on cohort studies for 10047 qualified participants from Ravansar County. Machine learning provides opportunities to improve discrimination through the analysis of complex interactions between broad variables. Among the chronic diseases in this cohort study, we have used three deep neural network models for diagnosis and prognosis of the risk of type 2 diabetes mellitus (T2DM) as a case study. Usually in Artificial Intelligence for medicine tasks, Imbalanced data is an important issue in learning and ignoring that leads to false evaluation results. Also, the accuracy evaluation criterion was not appropriate for this task, because a simple model that is labeling all samples negatively has high accuracy. So, the evaluation criteria of precession, recall, AUC, and AUPRC were considered. Then, the importance of variables in general was examined to determine which features are more important in the risk of T2DM. Finally, personality feature was added, in which individual feature importance was examined. Performing by Shapley Values, the model is tuned for each patient so that it can be used for prognosis of T2DM risk for that patient. In this paper, we have focused and implemented a full pipeline of Data Creation, Data Preprocessing, Handling Imbalanced Data, Deep Learning model, true Evaluation method, Feature Importance and Individual Feature Importance. Through the results, the pipeline demonstrated competence in improving the Diagnosis and Prognosis the risk of T2DM with personalization capability.
  7. Fulltext Cytotoxicity and Proapoptotic Effects of Allium atroviolaceum Flower Extract by Modulating Cell Cycle Arrest and Caspase-Dependent and p53-Independent Pathway in Breast Cancer Cell Lines
    Khazaei S, Abdul Hamid R, Ramachandran V, Mohd Esa N, Pandurangan AK, Danazadeh F, et al.
    Evid Based Complement Alternat Med, 2017;2017:1468957.
    PMID: 29250124 DOI: 10.1155/2017/1468957
    Breast cancer is the second leading cause of cancer death among women and despite significant advances in therapy, it remains a critical health problem worldwide. Allium atroviolaceum is an herbaceous plant, with limited information about the therapeutic capability. We aimed to study the anticancer effect of flower extract and the mechanisms of action in MCF-7 and MDA-MB-231. The extract inhibits the proliferation of the cells in a time- and dose-dependent manner. The underlying mechanism involved the stimulation of S and G2/M phase arrest in MCF-7 and S phase arrest in MDA-MB-231 associated with decreased level of Cdk1, in a p53-independent pathway. Furthermore, the extract induces apoptosis in both cell lines, as indicated by the percentage of sub-G0 population, the morphological changes observed by phase contrast and fluorescent microscopy, and increase in Annexin-V-positive cells. The apoptosis induction was related to downregulation of Bcl-2 and also likely to be caspase-dependent. Moreover, the combination of the extract and tamoxifen exhibits synergistic effect, suggesting that it can complement current chemotherapy. LC-MS analysis displayed 17 major compounds in the extract which might be responsible for the observed effects. Overall, this study demonstrates the potential applications of Allium atroviolaceum extract as an anticancer drug for breast cancer treatment.
  8. Fulltext Prevalence of HIV, Hepatitis B and C Virus Co-infections among Iranian High-Risk Groups: A Systematic Review and Meta-Analysis
    Hashiani AA, Sadeghi F, Ayubi E, Rezaeian S, Moradi Y, Mansori K, et al.
    Malays J Med Sci, 2019 May;26(3):37-48.
    PMID: 31303849 DOI: 10.21315/mjms2019.26.3.3
    Co-infection with hepatitis B and C among HIV infected patients are prevalent among high-risk populations. This meta-analysis aimed to estimate the prevalence of HIV, HCV and HBV co-infections among high-risk populations in Iran. We systematically searched the national and international electronic databases until 2016. The primary outcome was the prevalence of HIV, HBV, HCV and HIV co-infections in different high-risk populations in Iran. All English and Persian studies conducted on Iranian high-risk groups were included in the study. The review was reported based on PRISMA guidelines and data were analysed at 95% confidence level using random effect models. Overall, 916 relevant papers were recognised and 14 articles were included in the meta-analysis. The pooled estimates of HBV/HCV, HCV/HIV, HBV/HIV and HBV/HCV/HIV were 1.3% (95%CI: 0.5-2.1), 16.3% (95%CI: 1.1-31.6), 0.5% (95%CI: 0-1.4) and 0.5% (95%CI: 0.2-0.8), respectively. Based on subgroup analysis, there was a higher proportion of all co-infections from the years 2010-2016 as compared to that of the years 2003-2009. Our results highlighted that HCV/HIV co-infection in Iranian high-risk groups including injection drug users (IDUs) and prisoners is common. In addition, the increasing trend of coinfections should be considered alarming for policymakers.
  9. Fulltext Estimating global injuries morbidity and mortality: methods and data used in the Global Burden of Disease 2017 study
    James SL, Castle CD, Dingels ZV, Fox JT, Hamilton EB, Liu Z, et al.
    Inj Prev, 2020 Oct;26(Supp 1):i125-i153.
    PMID: 32839249 DOI: 10.1136/injuryprev-2019-043531
    BACKGROUND: While there is a long history of measuring death and disability from injuries, modern research methods must account for the wide spectrum of disability that can occur in an injury, and must provide estimates with sufficient demographic, geographical and temporal detail to be useful for policy makers. The Global Burden of Disease (GBD) 2017 study used methods to provide highly detailed estimates of global injury burden that meet these criteria.

    METHODS: In this study, we report and discuss the methods used in GBD 2017 for injury morbidity and mortality burden estimation. In summary, these methods included estimating cause-specific mortality for every cause of injury, and then estimating incidence for every cause of injury. Non-fatal disability for each cause is then calculated based on the probabilities of suffering from different types of bodily injury experienced.

    RESULTS: GBD 2017 produced morbidity and mortality estimates for 38 causes of injury. Estimates were produced in terms of incidence, prevalence, years lived with disability, cause-specific mortality, years of life lost and disability-adjusted life-years for a 28-year period for 22 age groups, 195 countries and both sexes.

    CONCLUSIONS: GBD 2017 demonstrated a complex and sophisticated series of analytical steps using the largest known database of morbidity and mortality data on injuries. GBD 2017 results should be used to help inform injury prevention policy making and resource allocation. We also identify important avenues for improving injury burden estimation in the future.

  10. Fulltext Mapping 123 million neonatal, infant and child deaths between 2000 and 2017
    Burstein R, Henry NJ, Collison ML, Marczak LB, Sligar A, Watson S, et al.
    Nature, 2019 Oct;574(7778):353-358.
    PMID: 31619795 DOI: 10.1038/s41586-019-1545-0
    Since 2000, many countries have achieved considerable success in improving child survival, but localized progress remains unclear. To inform efforts towards United Nations Sustainable Development Goal 3.2-to end preventable child deaths by 2030-we need consistently estimated data at the subnational level regarding child mortality rates and trends. Here we quantified, for the period 2000-2017, the subnational variation in mortality rates and number of deaths of neonates, infants and children under 5 years of age within 99 low- and middle-income countries using a geostatistical survival model. We estimated that 32% of children under 5 in these countries lived in districts that had attained rates of 25 or fewer child deaths per 1,000 live births by 2017, and that 58% of child deaths between 2000 and 2017 in these countries could have been averted in the absence of geographical inequality. This study enables the identification of high-mortality clusters, patterns of progress and geographical inequalities to inform appropriate investments and implementations that will help to improve the health of all populations.
  11. Fulltext Global, Regional, and National Cancer Incidence, Mortality, Years of Life Lost, Years Lived With Disability, and Disability-Adjusted Life-Years for 29 Cancer Groups, 1990 to 2017: A Systematic Analysis for the Global Burden of Disease Study
    Global Burden of Disease Cancer Collaboration, Fitzmaurice C, Abate D, Abbasi N, Abbastabar H, Abd-Allah F, et al.
    JAMA Oncol, 2019 Dec 01;5(12):1749-1768.
    PMID: 31560378 DOI: 10.1001/jamaoncol.2019.2996
    IMPORTANCE: Cancer and other noncommunicable diseases (NCDs) are now widely recognized as a threat to global development. The latest United Nations high-level meeting on NCDs reaffirmed this observation and also highlighted the slow progress in meeting the 2011 Political Declaration on the Prevention and Control of Noncommunicable Diseases and the third Sustainable Development Goal. Lack of situational analyses, priority setting, and budgeting have been identified as major obstacles in achieving these goals. All of these have in common that they require information on the local cancer epidemiology. The Global Burden of Disease (GBD) study is uniquely poised to provide these crucial data.

    OBJECTIVE: To describe cancer burden for 29 cancer groups in 195 countries from 1990 through 2017 to provide data needed for cancer control planning.

    EVIDENCE REVIEW: We used the GBD study estimation methods to describe cancer incidence, mortality, years lived with disability, years of life lost, and disability-adjusted life-years (DALYs). Results are presented at the national level as well as by Socio-demographic Index (SDI), a composite indicator of income, educational attainment, and total fertility rate. We also analyzed the influence of the epidemiological vs the demographic transition on cancer incidence.

    FINDINGS: In 2017, there were 24.5 million incident cancer cases worldwide (16.8 million without nonmelanoma skin cancer [NMSC]) and 9.6 million cancer deaths. The majority of cancer DALYs came from years of life lost (97%), and only 3% came from years lived with disability. The odds of developing cancer were the lowest in the low SDI quintile (1 in 7) and the highest in the high SDI quintile (1 in 2) for both sexes. In 2017, the most common incident cancers in men were NMSC (4.3 million incident cases); tracheal, bronchus, and lung (TBL) cancer (1.5 million incident cases); and prostate cancer (1.3 million incident cases). The most common causes of cancer deaths and DALYs for men were TBL cancer (1.3 million deaths and 28.4 million DALYs), liver cancer (572 000 deaths and 15.2 million DALYs), and stomach cancer (542 000 deaths and 12.2 million DALYs). For women in 2017, the most common incident cancers were NMSC (3.3 million incident cases), breast cancer (1.9 million incident cases), and colorectal cancer (819 000 incident cases). The leading causes of cancer deaths and DALYs for women were breast cancer (601 000 deaths and 17.4 million DALYs), TBL cancer (596 000 deaths and 12.6 million DALYs), and colorectal cancer (414 000 deaths and 8.3 million DALYs).

    CONCLUSIONS AND RELEVANCE: The national epidemiological profiles of cancer burden in the GBD study show large heterogeneities, which are a reflection of different exposures to risk factors, economic settings, lifestyles, and access to care and screening. The GBD study can be used by policy makers and other stakeholders to develop and improve national and local cancer control in order to achieve the global targets and improve equity in cancer care.

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