Dengue fever is one of the major health problems in tropical and subtropical areas throughout the world. The causative agent of dengue fever is the dengue virus which is an enveloped single stranded RNA virus belongs to the family Flaviviridae and has five distinct serotypes (DENV-1, DENV-2, DENV-3, DENV-4 and DENV-5). Dengue virus is transmitted to human via bite of Aedes aegypti and Aedes albopictus mosquitoes. The clinical symptoms of dengue fever ranging from mild to severe fonn as dengue hemorrhagic fever and dengue shock syndrome. Pakistan is dengue endemic since 1994 but from 2006, Pakistan faced the worst condition regarding dengue in which thousands of people affected by the disease and hundreds of people lost their lives. DENV-2, DENV-3 and DENV-1 are the prevalent serotypes in Pakistan. Common diagnostic techniques are being used in Pakistan such as enzyme-linked immunosorbent assay, polymerase chain reaction and rapid diagnostic tests, while differential diagnosis, limitations of diagnostic methods and poor health care system are the real challenges in dengue diagnosis. Favorable climatic conditions, unplanned urbanization, travelling etc., are major factors responsible for dengue epidemics in Pakistan. This presentation provides update about dengue circumstances in Pakistan and also describes the way how to improve dengue situation in Pakistan.

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Objective: To investigate 33 organic extracts of mangrove plants for: antiparasitic, anticancer, and antibacterial activities, as well as their ability to inhibit the activity of the α-glucosidase enzyme. Methods: Leaves from all different plant mangrove species located in five mangrove zones of the Pacific coast of Panama were collected according to standard procedures. Qualitative phytochemical analysis of the organic extracts was performed by thin layer chromatography. The antiparasitic activity against Plasmodium falciparum, Trypanosoma cruzi and Leishmania donovani, toxicity against Artemia salina, anticancer activity in MCF-7 cell line, and antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa of all organic extract were investigated according protocols stablished in our institution. Finally, the ability to inhibit the enzymatic activity of α-glucosidase was evaluated by monitoring the hydrolysis of p-nitrophenyl α-D- glucopyranoside. Results: Thirty-three different samples belonging to nine different species of vascular plants with seeds of true mangroves were collected. Triterpenoids, phenolics, and tannins were the main groups of compounds found in the sampled mangroves. Saponins, quinones, and coumarins were found in less than 50% of the samples. Laguncularia racemosa showed moderate activity against Plasmodium falciparum. None of the extracts presented anticancer activity. Rhizophora mangle exhibited potent activity against Staphylococcus aureus and Bacillus subtilis [(90.41±7.33)% and (96.02±6.14)% of inhibition]; Avicennia germinans and Conocarpus erectus had activity against Escherichia coli [(71.17±6.15)% and (60.60±5.13)% of inhibition, respectively]. About 60% of the mangroves showed α-glucosidase inhibitory activity. In particular, extracts from Laguncularia racemosa, Pelliciera rhizophorae, Conocarpus erectus, Mora oleifera, and Tabebuia palustris species showed α-glucosidase inhibitory potential, with IC₅₀ values of (29.45±0.29), (20.60±0.70), (730.06±3.74), (25.59±0.37), and (853.39±5.30) μg/mL, respectively. Conclusions: Panamanian mangroves are mainly a promising potential source of hypoglycemic compounds, specifically α-glucosidase inhibitors. These results highlight the therapeutic virtues of extracts from American mangrove plants.

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Madhuca longifolia (M. longifolia) is also known as Mahua belonging to the family sapoteace family. M. longifolia is used in traditional and folklore system of medicine widely across India, Nepal, and Sri Lanka for its various pharmacological properties as in snake bites and in diabetes. Phytochemicals studies documented the different bioactive constituents, namely, glycosides, flavonoids, terpenes and saponins. The pharmacological studies proved that it possess wide range of biological activities such as antiulcer, antiinflammatory, antioxidant and antidiabetic activities. The toxicity studies reveal its non-toxic effect even at larger doses. Thus M. longifolia can be considered as a therapeutic agent for specific diseases. Scientific investigation on various isolated bioactive components and its efficacy on diseases proved the future usefulness of different species of Madhuca. This review summarizes the phytochemical, pharmacological, medicinal and non-medicinal uses of M. longifolia. Further exploration on M. longifolia for its therapeutic potential is however required for depth traditional knowledge.

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Objective: To determine Toxoplasma gondii (T. gondii) as a cause of morbidity and mortality in human immunodeficiency virus (HIV) infected patients by ELISA method and to investigate toxoplasmosis-associated risk factors. Methods: This cross sectional study was conducted with 385 patients with HIV/AIDS referred to the behavior disease consultation center in Kermanshah, Iran between May 2012 and June 2013. Anti-Toxoplasma IgG and IgM antibodies were measured by using the ELISA method. Also, the relationship between the infection and age, gender, education, occupation, antiretroviral status, CD4+ cell counts and some other factors of the patients were assessed. P<0.05 was considered statistically significant. Results: Among the HIV-positive patients, 40.8% (157/358) and 2.6% (10/358) patients were positive for anti-T. gondii IgG and IgM antibodies, respectively. The mean CD4+ count in the patients with HIV was 327 cells/μL. A significant correlation was observed between the toxoplasmosis infections with the age group (P<0.05). Nevertheless, no statistically significant difference was found between gender, living area, educational level, occupational, marital status, antiretroviral status, prophylaxis, CD4+ cell count and toxoplasmosis. Conclusions: This study revealed that the patients with HIV infection were at the high risk of developing toxoplasmosis disease especially those patients who do not receive antiretroviral therapy and prophylaxis. The development and use of the effectiveness-based guidelines for controlling and the prevention reactivation of the latent T. gondii infection is required.

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Objective: To determine the seroprevalence of varicella zoster virus (VZV) antibodies among the population residing in the Colombo district of Sri Lanka. Methods: A cross-sectional population-based study was conducted which included 1 258 participants. Blood samples were collected and questionnaires administered to obtain sociodemographic information and history of varicella and/or herpes zoster. Serum samples were assayed for VZV IgG antibodies using a commercial enzyme-linked immunosorbent assay kit. Results: Overall, the seroprevalence was 54.2% (95% CI = 51.5% 57.0%). Children below 1 year of age were seronegative, and only about 20.0% of children between 1 and 10 years of age were seropositive. Seropositivitiy increased with age and by the age of 40 years 74.3% were seropositive. Among women of childbearing age, the overall seroprevalence was about 62.0% (95% CI = 57.7%-66.1%) but was low 37.0% in the 15-19 age group. Conclusion In this population, 45.8% lacked natural immunity against varicella. Of women of childbearing age, 39.9% lacked immunity and in the subgroup of women 15-19 years of age, 63.0% women lacked immunity. In light of the country's success with the control and high coverage of other vaccine preventable diseases and that the vaccine is available in the private sector, the inclusion of varicella vaccine in the national immunization program may be considered.

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Objective: To investigate the effects of a dry aqueous extract of Notobasis syriaca (N. syriaca) on lipopolysaccharide (LPS)-induced inflammation in rats. Methods: Rats were fed the dried extract [500 mg/(kgod)] for three consecutive days and then were intraperitoneally injected with LPS (1 mg/kg). Two hours after LPS injection, rats were sacrificed and blood and brain regions were collected. Inflammatory mediators’ levels in plasma and homogenates of brain regions were determined by ELISA. Results: Pretreatment with the N. syriaca extract resulted in significant anti-inflammatory effects (P<0.05), including: i) attenuated LPS-induced hypothermia; ii) decreased hypothalamus and hippocampus prostaglandin E₂ levels in the LPS- treated rats; and, iii) reduced hypothalamus and hippocampus interleukin-6 and tumor necrosis factor- α levels in the LPS-treated rats. Conclusions: These results suggest that N. syriaca possesses anti-inflammatory properties. Thus, it is possible that long-term consumption of this plant may result in beneficial pharmacological effects.

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Objective: To investigate the free radical scavenging, antidiabetic, kinetics and cytotoxic potentials of flavonoids extract of Dicoma anomala root by using standard methods. Methods: Antioxidant activity of the flavonoids was investigated at scavenging free radicals such as 1,1- diphenyl-2-picrylhydrazyl, nitric oxide, hydroxyl radical, reducing capabilities, 2,2-azino- bis (3-ethylbenzothiazoline-6) sulfonic acid as well as metal chelating capability at different concentrations (0.125-1.000 mg/mL) while the antidiabetic activity was evaluated via the inhibition and kinetics of carbohydrate digestive enzymes including, alpha glucosidase, sucrase and maltase. Brine shrimp lethality assay was also employed to examine the cytotoxic effects of the extract by using different range of concentrations (0.125-2.000 mg/mL). Results: The study revealed the best antioxidant activity of the extract in 1,1-diphenyl-2-picrylhydrazyl, 2,2-azino-bis (3-ethylbenzothiazoline-6) sulfonic acid and nitric oxide having IC₅₀ values of (386.90±4.91), (736.00±38.12), (629.30±9.62) μg/mL respectively compared with quercetin (standard) with IC₅₀ [(522.20±12.38), (1 021.00±15.61) and (1 075.00±29.35) μg/mL] respective values while it was insignificantly (P>0.05) at par with quercetin for reducing power. Similarly, the extract exhibited a moderate inhibition of alpha glucosidase (43.1%), sucrase (33.4%) and maltase (29.9%) activities which were significantly (P<0.05) better than acarbose (18.4%, 12.7% and 24.9% respectively) although acarbose (46.1%) inhibited the higher activity of alpha amylase than the extract (13.7%). The kinetics of mode of inhibition of alpha amylase, alpha glucosidase, sucrase and maltase by flavonoids extract of Dicoma anomala revealed an uncompetitive, non-competitive, competitive and non-competitive inhibition respectively. The result of the lethality assay showed a potent cytotoxic effect of the flavonoids with LC₅₀ value 0.510 mg/mL. Conclusions: The results obtained from this study are suggestive of the antioxidative, antidiabetic and cytotoxic potentials of flavonoids root extract of Dicoma anomala.

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Objective: To determine the breadth of Zika virus (ZIKV)-associated brain anomalies in neonates and adults. Methods: Systematic review was conducted according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement using electronic databases ScienceDirect, Pubmed, Medline, Scopus, and Global Health Library. Only those research articles, case studies, case-control studies, case-cohort studies, cross-sectional studies, and organizational survey reports were included in the study that reported any fetal outcomes for pregnant women who had infected with ZIKV during the gestational period and ZIKV-related neurological complications in adults as well. Results: Out of total 72 retrieved articles, 50 met the inclusion criteria. We estimated a significant increase in incidence of neural abnormalities such as Guillain-Barre syndrome and microcephaly in the regions that are experiencing ZIKV outbreaks. Other neurological malformations found in ZIKV patients include hydrancephaly/hydrops fetalis, myasthenia gravis, meningoencephalitis and myelitis. Conclusion: Our systematic analysis provides the broad spectrum of neurological malformations in ZIKV infected patients and these data further support the causal link of ZIKV with neurological disorders.

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Objective: To study the injury effect and molecular mechanism of high glucose on myocardial cells. Methods: Myocardial cells H9c2 were cultured and divided into the control group treated with DMEM containing 5.5 mmol/L glucose, the high glucose group treated with DMEM containing 35 mmol/L glucose, and the N-acetylcysteine (NAC) group pre-treated with 1 000 μmol/L NAC and treated with DMEM containing 1 000 μmol/L NAC and 35 mmol/L glucose. The production of ROS and the expression of mitochondria pathway apoptosis molecules in cells as well as the contents of collagen and collagen metabolism molecules were measured. Results: After 8 h, 16 h and 24 h of treatment, ROS RFU as well as Bax, CytC, Caspase-3 and Caspase-9 protein expression in cells and Col-I, Col-III, PINP and PIIINP protein levels in culture medium of high glucose group were higher than those of control group, Bcl-2 protein expression were lower than those of control group, but CTX-I protein levels in culture medium were not significantly different from those of control group; after 24 h of treatment, Bax, CytC, Caspase-3 and Caspase-9 protein expression in cells as well as Col-I, Col-III, PINP and PIIINP protein levels in culture medium of NAC group were lower than those of high glucose group whereas Bcl-2 protein expression was higher than that of high glucose group. Conclusions: High glucose can induce myocardial cell apoptosis, increase collagen synthesis and accelerate interstitial fibrosis by increasing the production of reactive oxygen species.

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Objective: To investigate the effect of the prostaglandin E1 analogue misoprostol on oxidative stress and neurodegeration caused by subcutaneous rotenone administration in rats. Methods: Rotenone was administered in a dose of 1.5 mg/kg every other day for 2 weeks. Starting from the 1st day of rotenone injection, rats were subcutaneously treated with misoprostol at doses of 10, 100 or 1 000 μ g/kg. Rats were evaluated for brain lipid peroxidation (malondialdehyde: MDA), reduced glutathione (GSH), nitric oxide (NO) levels, and paraoxonase-1 (PON-1) activity. The concentrations of the anti-apoptotic protein B cell/lymphoma-2 (Bcl-2) were determined in the striatum. Histopathologic examination and the expression of inducible nitric oxide synthase (iNOS) in the cerebral cortex and striatum were also performed. Results: Compared with the vehicle-treated group, rotenone caused a significant increase in brain lipid proxidation (MDA) by 61% (P<0.05) accompanied by an increase in NO by 73.1% (P<0.05) and a decrease in GSH concentration by 29.4% (P<0.05). In addition, brain PON-1 activity significantly decreased by 63.0% (P<0.05) and striatal Bcl-2 significantly decreased by 27.9% (P<0.05) with respect to the corresponding control value. Brain sections from rotenone treated rats showed extensive dark pyknotic and apoptotic nuclei in neurons, shrunken cytoplasm and perineuronal vacuolation. Rotenone also caused pronounced expression of iNOS in the cerebral cortex and striatum. Treatment with misoprostol at doses of 100 and 1 000 μ g/kg resulted in decreased brain MDA (by 16.5%-23.0%) (P<0.05) and NO levels (by 37.1%-40.7%) (P<0.05) and increased GSH concentrations (by 18.8%-30.1%) (P<0.05). PON-1 activity was significantly increased by 80.0%-114.8% (P<0.05) by misoprostol at 100 and 1 000 μ g/kg, respectively. In addition, misoprostol treatment restored striatal Bcl-2 concentrations to its normal value. Misoprostol treatment resulted in markedly reduced brain injury and decreased iNOS expression in the cerebral cortex and striatum of rotenone intoxicated rats. Conclusions: These data suggest that misoprostol prevents the rotenone-induced neurodegeneration in rat brain by reducing brain oxidative stress.

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Objective: To better understand the economic burden of patients with epilepsy receiving outpatient services in China and to analyze potential factors influencing epileptic economic burden through a cross-sectional study. Methods: Using a self-designed questionnaire, we collected information retrospectively from 754 patients with epilepsy evaluated in neurology clinics in Shanghai Municipality, Shanxi Province and Sichuan Province. Descriptive analyses were used after cost variables were presented as logarithms, and multiple linear regressions were performed to explore influencing factors. Results: Fifty percent of the investigated patients experienced an epilepsy attack before the age of 15, and 51.3% had suffered from epilepsy for more than five years. In the past year, 87.9% of patients had visited different hospitals multiple times for evaluation (40.3%) and maintenance treatment (40.7%). The total economic burden of epilepsy was US$ 1 143.2. The average direct economic burden and indirect economic burden were US$ 939.0 and US$ 110.2, respectively. Multiple linear regressions showed that patients had to bear greater economic burden if they were hospitalized, using multiple antiepileptic drugs, experiencing illness for less than 5 years, in severe seizure index or active epilepsy with drug resistance, which was statistically significant. Totally only 14.3% of patients could get reimbursement in outpatient services. Conclusions: Patients with epilepsy must present to hospitals regularly for satisfactory prognosis, which results in economic burden. Patients bear greater economic burden, especially direct medical costs, if they are newly diagnosed, experience severe seizures, or undergo multiple drug treatments that require more frequent monitoring. However, current insurance policy for outpatient services do not help reduce economic burden of patients efficiently.

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Objective: To study the adverse effects of advanced glycation end products (AGEs) on chondrocytes and the role of autophagy in this process. Methods: Chondrocytes were harvested from the human articular cartilage tissues in surgery. AGEs were administered during chondrocytes culture. The rapamycin was used to induce autophagy. The cell viability was determined by 3-[4,5-dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. The expression of tumor necrosis factor- α (TNF- a ) and nuclear factor- κ B (NF- κ B) was detected by quantitative real-time polymerase chain reaction. The reactive oxygen species (ROS) production and apoptosis of the chondrocytes were determined by fluorescent probe and flow cytometer, respectively. Results: The chondrocytes viability was significantly reduced after 12 h incubation with AGEs (P<0.01)). In contrast, rapamycin pretreatment increased the chondrocytes viability through autophagy. AGEs increased TNF- α and NF- κ B mRNA expression of chondrocytes and autophagy receded or proceeded the change. AGEs increased intracellular ROS accumulation and autophagy reversed the change. AGEs accelerated chondrocytes apoptosis and autophagy suspended apoptosis. Conclusions: Accumulation of AGEs may have an adverse role for chondrocytes by increasing TNF- α and NF- κ B expression, ROS accumulation and apoptosis; meanwhile, autophagy ameliorates the AGEs- induced adverse effects.

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Objective: To observe the expression and significance of fibronectin and metalloproteinase-3 (MMP-3) in patients with ankylosing spondylitis (AS). Methods: A total of 30 AS patients in our hospital and 30 healthy volunteers were selected in our study. Fibronectin and MMP-3 were measured and compared between these two groups. The AS group received sulfasalazine 2 g daily for 3 months. Bath ankylosing spondylitis disease activity index, bath ankylosing spondylitis functional index, bath ankylosing spondylitis metrology index, erythrocyte sedimentation rate and C-reactive protein were compared before and after treatment. Pearson's linear-correlation analysis was used to determine relationships between parameters. Results: Totally 28 patients in the AS group completed the study. Fibronectin and MMP-3 in peripheral blood of AS patients were evidently higher than that in the normal control group (P<0.05). After treated by sulfasalazine, the level of expressing Fibronectin and MMP-3 significantly decreased compared with baseline values (P<0.05). Pearson's linear-correlation analysis showed that serum fibronectin and MMP-3 level had a positive correlation with bath ankylosing spondylitis disease activity index global assessment, spine pain, night pain, general pain, erythrocyte sedimentation rate and C-reactive protein (P<0.05). Conclusions: The expression of fibronectin and MMP-3 in AS patients were significantly higher than that in the normal control group, and they all decreased significantly after treatment. It indicated that both fibronectin and MMP-3 were correlated closely with the onset of AS.

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Objective: To identify potential tumor markers for the development and recurrence of hepatocelullar carcinoma (HCC), this research studied the relationship between the expression of the tumor necrosis factor receptor-associated factor 4 (TRAF4) and tumor angiogenesis together with its survival time of HCC patients. Methods: The expressions of TRAF4, vascular endothelial growth factor and CD34 were performed upon 90 patients with curative liver resection between August 2006 and November 2009 by immunohistochemical method in locally advanced HCC and adjacent non-tumoral liver. The expression of TRAF4 was determined by the Spearman rank correlation. Their prognostic factors on disease free survival (DFS) and overall survival (OS) were guaranteed by Kaplan-Meier and Cox regression analyses. The detection of the levels of vascular endothelial growth factor and CD34 was fulfilled in 90 cases of HCC. Results: TRAF4 expression was both significantly higher in HCC than in surrounding non-tumor tissues (57.8% vs. 22.2 %; P<0.001) and significantly correlated with tumor size and tumor staging. High TRAF4 was correlated with reduced DFS rate (P=0.001) and overall OS rate (P<0.001) and were displayed in Kaplan-Meier survival analysis. Conclusions: TRAF4 is involved with multifarious clinicopathologic features. TRAF4 expression, as an independent adverse prognostic factor, DFS and OS in HCC, is associated with increased tumor angiogenesis. The combined detection of TRAF4 in locally advanced HCC is a trustworthy predictive factor for the tumor development and recurrence.

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