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Oxidative stress and NF-κB signaling plays a major role in pathogenesis of osteoarthritis. In the present study, we analyzed the potent role of carnosol against osteoarthritis in cells treated using monosodium iodoacetate (MIA) model through in vitro studies. MIA caused dose-dependent cell death and induced programmed cell death by increasing subG1 accumulation and caspase-3 expressions. MIA caused oxidative stress by increasing reactive oxygen species, lipid peroxidation and further induced NF-κB expression and down regulated Nrf-2 levels. Pre-treatment with carnosol significantly protected the cells by reducing the oxidative stress markers and improved the cell viability up to 98%. Further, carnosol down regulated NF-κB nuclear expression with a concomitant increase in Nrf-2 nuclear localization and up regulated the nuclear Nrf-2 levels. Carnosol also inhibited MIA-induced subG1 accumulation and caspase-3 activation. This study demonstrates that, carnosol might act as potent antioxidant and regulate MIA-induced oxidative stress, NF-κB signaling and programmed cell death by up regulating the Nrf-2 levels.

Sporadic Alzheimer's disease is characterised by cognitive impairments and associated with cerebrometabolic anomaly and dementia which are burdensome for the old age population and their caregivers worldwide. A safe and effective treatment is essentially required for its prevention and cure. γ-Oryzanol (OZ) is reported for anti-oxidative, anti-inflammatory, anticancer, anti-hyperlipidemic, and anti-diabetic effects in various preclinical studies. In silico studies showed similar binding interactions with acetylcholinisterase like Donepezil (DONO). In vitro DPPH assay, AchE activity inhibition assay and cell viability assay on SH-SY5Y cell line confirmed that OZ has good free radical scavenging and cholinesterase inhibitory activity as well as safety profile. OZ was evaluated in vivo for its effect in streptozotocin (STZ) induced sporadic Alzheimer's disease in rats. Maze tests reflected improvement in spatial cognitive behavior indicated by reduction in working memory and reference memory error. OZ showed anti-AchE, anti-inflammatory and antioxidative effects. OZ showed effective anti-proinflammatory effects in cerebral milieu as indicated by significant reduction in active glial cells and found to improve synaptic connectivity. Thereby OZ reflected protection of the cerebral architecture against STZ induced damage. Thus, OZ exhibits its candidature as a therapeutic moiety to improve cognitive behavior in neurodegenerative disorders.

Silibinin is known to display high efficacy against cancer cells and for hepatic protection. Metformin, a well-known antidiabetic agent, has recently been reported to inhibit cancer. In the present study, we investigated the effect of metformin on silibinin-induced programmed cell death in cervical cancer cells (C-33A). MTT assay and Western blot assays were performed to quantify cell viability and the expression of signaling proteins, respectively. Combined treatment with metformin and silibinin decreased cell survival in synergistic manner in C-33A cells at a dose that did not affect nonmalignant cells (HUVECs). Silibinin and metformin increased PTEN and AMPK expression in C-33A cells, respectively. Combined treatment caused a greater increase in the expression of activated caspase-3 or AIF, indicating apoptosis. Combined treatment with silibinin and metformin may induce programmed cell death of human cervical cancer cells at a dose that does not affect HUVECs. This finding reveals a potential therapeutic strategy of cervical cancer.

The perinucleolar region represents a special nuclear compartment involved in the cell malignancy and the perinucleolar heterochromatin reflects the presence of silent genes. The present study was undertaken to provide complementary and missing information on the perinucleolar heterochromatin in differentiating neutrophils in the bone marrow of patients with the chronic myeloid leukemia. That lineage is a very convenient model because of the increased number of granulocytic precursors that is satisfactory for size as well as optical density measurements in single cells. Moreover, the differentiation stages of neutrophils are well defined and easily identified. According to diameter measurements the enlarged width of the perinucleolar heterochromatin shell accompanied the decreasing nucleolar size in advanced stages of the cell differentiation. Such trend was not influenced by the anti-leukemic therapy with imatinib. Thus the increasing size of the perinucleolar heterochromatin shell with silent genes might reflect the genomic stability of the perinucleolar region during the cell differentiation. On the other hand, the increased perinucleolar heterochromatin condensation after the specific anti-leukemic therapy with imatinib indicated a "premature terminal differentiation" of leukemic neutrophils.

This study investigated the effects of sericin-derived oligopeptides on natural killer (NK) activity. In vitro exposure of human peripheral blood mononuclear cells with sericin-derived oligopeptides resulted in an augmentation of NK cell activity against K562 target cells and the effects appeared to be dose-related. Experiments designed to examine whether enhanced NK activity was due to direct or indirect activation of NK cells revealed that sericin oligopeptides did not induce activity of purified NK cells, and that sericin oligopeptides augmented NK activity indirectly by inducing the production of IL-2 and IFN-γ cytokines. In in vivo experimentation where mice were orally administered with sericin oligopeptides and splenic mononuclear cells tested against YAC-1 target cells, significant increase in NK activity was obtained compared to control mice. Elevated levels of IL-2 were also evident in all oligopeptides-treated groups. As demonstrated both in vitro and in vivo, these results indicate that sericin-derived oligopeptides have efficient NK-enhancing activity and suggest the potential therapeutic applications of such oligopeptides for functional improvement of NK cells, and possibly for treatment of tumor and infectious diseases in which NK activity contributes to host defense.

Health status is determined by the balance of oxidants and antioxidants which protects healthy cells against the threat of internal and external risk factors. Antioxidants such as ascorbate (vitamin C, ascorbic acid) are of fundamental importance in this respect. Ascorbate neutralizes potential damage caused by cellular oxidative stress which may be the greatest risk of damage to healthy tissue. Cellular oxidative stress is mediated by external factors (e.g. psychological stress, physical exertion, drugs, various diseases, environmental pollution, preservatives, smoking, and alcohol) and internal factors (products of cellular metabolism including reactive oxygen species). When the products of oxidative stress are not sufficiently neutralized, healthy cells are at risk for both mitochondrial and DNA damage. In the short term, cell function may deteriorate, while an increased production of proinflammatory cytokines over time may lead to the development of chronic inflammatory changes and diseases, including cancer. Although pharmaceutical research continues to bring effective chemotherapeutic agents to the market, a limiting factor is often the normal tissue and organ toxicity of these substances, which leads to oxidative stress on healthy tissue. There is increasing interest and imperative to protect healthy tissues from the negative effects of radio-chemotherapeutic treatment. The action of ascorbate against the development of oxidative stress may justify its use not only in the prevention of carcinogenesis, but as a part of supportive or complementary therapy during treatment. Ascorbate (particularly when administered parentally at high doses) may have antioxidant effects that work to protect healthy cells and improve patient tolerability to some toxic radio-chemotherapy regimens. Additionally, ascorbate has demonstrated an immunomodulatory effect by supporting mechanisms essential to anti-tumor immunity. Intravenous administration of gram doses of vitamin C produce high plasma levels immediately, but the levels drop rapidly. Following oral vitamin C administration, plasma levels increase slowly to relatively low values, and then gradually decay. With an oral liposomal formulation, significantly higher levels are attainable than with standard oral formulations. Therefore, oral administration of liposomal vitamin C appears to be an optimal adjunct to intravenous administration. In this review, the basic mechanisms and clinical benefits of ascorbate as an antioxidant that may be useful as complementary therapy to chemotherapeutic regimens will be discussed.

Background: Oral and topical nonsteroidal anti-inflammatory drugs (NSAIDs), analgesics and intra-articular corticosteroid injections are the recommended first line of treatment for knee osteoarthritis (OA); however, they have serious side effects. Platelet-rich plasma (PRP) has been posited as an effective and safer alternative treatment for knee OA. Hitherto, there is only one study comparing the effectiveness of PRP against an NSAID. Aim of the study: The aim of this study was to determine the effectiveness of PRP against celecoxib in the treatment of early knee OA. Methods: 60 patients with knee OA grade II and III were randomly alocated in two groups. Group 1 received one injection of autologous PRP in each affected knee, with a reinjection after 15 days; Group 2 received 200 mg of oral celecoxib each 24 h for a year. Visual Analogue Scale (VAS), total Western Ontario and McMaster Universities Arthritis Index (WOMAC) and WOMAC subscales for pain, stiffness and function were measured at baseline and at 1, 3, 6 and 12 months after the start of the treatment. Results: At the end of the study PRP was significantly better than celecoxib (p < 0.05) in improving VAS (40.40%), total WOMAC (58.95%) and WOMAC subscales of pain (50.60%), stiffness (34.13%) and function (51.90%). Significant differences remained after adjusting for age, sex or knee OA grade II or III. Conclusions: Intra-articular PRP is significantly better than celecoxib in improving pain, function and stiffness in early knee OA. This significant difference is independent of age, sex or knee OA grade II or III.

The most common cause of dementia in the elderly is Alzheimer disease (AD). In Europe, AD is a leading cause of death. The prevalence of this disease in developed countries is increasing because of very significant shifts in life expectancy and demographic parameters. AD is characterized by progressive cognitive impairment, resulting from dysfunction and degeneration of neurons in the limbic and cortical regions of the brain. Two prominent abnormalities in the affected brain regions are extracellular deposits of β-amyloid, and intracellular aggregates of tau protein in neurofibrillary tangles. The role of these features in AD pathogenesis and progression is not yet completely elucidated. Research over the last decade has revealed that the activation of cell cycle machinery in postmitotic neurons is one of the earliest events in neuronal degeneration in AD. Here we summarize evidence to support the hypothesis that cell cycle alterations occur in cells other than neurons in AD sufferers. Immortalized lymphocytes from AD patients have show an enhanced rate of proliferation associated with G1/S regulatory failure induced by alterations in the cyclin/CDK/pRb/E2F pathway. In addition, these cells have a higher resistance to serum deprivation-induced apoptosis. These neoplastic-like features, cell cycle dysfunction and impaired apoptosis can be considered systemic manifestations of AD disease.

A small molecule EGFR inhibitor, 4-(2-(3-(4-(4-(trifluoromethyl)phenyl)thiazol-2-yl)ureido)vinyl)-1,2-phenylene diacetate ( CIU1) was designed in silico by using caffeic scaffold as core structure. The designed compound showed anti-proliferative action against different solid tumor cell lines, particularly metastatic breast cancer cells. CIU1 inhibited the growth of EGFR-overexpressing MDA-MB-468 triple-negative breast cancer cells and wild-type non-small-cell lung cancer H460 cells with IC₅₀ values of 8.96 μM and 12.98 μM, respectively, these anti-proliferative effects of CIU1 were comparable to gefitinib (a specific EGFR inhibitor) or lapatinib (a dual EGFR and HER2 tyrosine kinase inhibitor). Interestingly CIU1 effectively inhibited the invasive hormone-dependent MCF-7 cancer cells with an IC₅₀ 2.34 μM. The immunoblot analyses revealed that CIU1 induced programmed cell death and suppressed EGFR expression in EGFR-overexpressing breast cancer (MDA-MB468) and lung cancer (PC-9) cells. The findings substantiated our design strategy and demonstrated the potential of CIU1 as new lead for further optimization in the development of anticancer drugs against advanced solid tumors.

Fruit of Terminalia chebula Retz. (Combretaceae) has male contraceptive folk medicine reputation but its molecular aspect regarding hypotesticular activity is still in dark. The study focused the hypotesticular efficacy of the most potent fraction out of n-hexane, chloroform and ethyl acetate fractions of hydro-methanolic (3:2) extract of Terminalia chebula in connection with male herbal contraceptive development.
Treatment with above fractions of Terminalia chebula showed a significant diminution in the activities of androgenic key enzymes (Δ5, 3β-HSD, 17β-HSD) and inhibition in serum testosterone level in compare to the control. Significant up regulation of testicular Bax gene and down regulation of Bcl-2 gene indicated the hypotesticular activity of these fractions. Flow-cytometric study focused a significant diminution in sperm viability and sperm mitochondrial status after the treatment with different fractions. Out of these, ethyl acetate fraction showed most promising hypotesticular effect without impairing any toxicity in general which highlighted that the fraction may contains antitesticular agent(s) in threshold levels compare to other fractions as it decreases spermiological, testicular genomic sensors and elevates sperm apoptotic sensors that may lead to male contraception.

Takayasu disease belongs to the group of autoimmune vasculitis which most often affects the aorta and its branches. It is rare, and it mainly affects young women. Recent epidemiologic studies suggest that Takayasu arteritis is being increasingly recognized in Europe. The first symptoms are non-specific and an early diagnosis is difficult and requires clinical awareness and suspicion. Patients with Takayasu arteritis often present increased inflammatory markers, including C-reactive protein and erythrocyte sedimentation rate, but systemic inflammatory response does not always show a positive correlation with inflammatory activity in the vessel wall. Therefore, imaging studies play a principal role in diagnosis and control of the disease. Glucocorticoids remain the most effective and serve as a cornerstone first line treatment. Immunosuppressive drugs play an important role as well, and biological therapy is increasingly being included in the treatment. This article describes the epidemiology, pathophysiology, diagnostics and treatment of this rare disease, so as to alert clinicians because disease left untreated can lead to narrowing and even closure of vital blood vessels. The most common Takayasu arteritis complications include pulmonary thrombosis, aortic regurgitation, congestive heart failure, cerebrovascular events, vision degeneration or blindness, and hearing problems.

Lung cancer (LC) incidence represents 11.5% of all new cancers, resulting in 1.72 million deaths worldwide in 2015. With the aim to investigate the capability of the electronic nose (e-nose) technology for detecting and differentiating complex mixtures of volatile organic compounds in biofluids ex-vivo, we enrolled 50 patients with suspected LC and 50 matching controls. Tissue biopsy was taken from suspicious lung mass for histopathological evaluation and blood, exhaled breath, and urine samples were collected from all participants and qualitatively processed using e-nose. Odor-print patterns were further analysed using the principal component analysis (PCA) and artificial neural network (ANN) analysis. Adenocarcinoma, non-small cell LC and squamous cell carcinoma were the predominant pathological types among LC patients. PCA cluster-plots showed a clear distinction between LC patients and controls for all biological samples; where the overall success ratios of classification for principal components #1 and #2 were: 95.46, 82.01, and 91.66% for blood, breath and urine samples, respectively. Moreover, ANN showed a better discrimination between LC patients and controls with success ratios of 95.74, 91.67 and 100% for blood, breath and urine samples, respectively. The e-nose is an easy noninvasive tool, capable of identifying LC patients from controls with great precision.

Recent literature evidence indicates the potential use of chokeberry preparations in the prevention and treatment of some chronic noncommunicable diseases. The aim of the present study was to evaluate the effects of the three months oral chokeberry juice supplementation in type 2 diabetic patients, as well as its influence on hematological parameters and certain parameters of the renal dysfunction. The study was designed as an open-label trial, which included 35 patients who have received the herbal supplement, polyphenol-rich chokeberry juice (150 ml/day, three times a day for 50 ml), in addition to their standard therapy. Chokeberry juice as a rich source of polyphenol compounds could be an effective preventive and therapeutic agent in diabetes mellitus type 2. Hematological and biochemical parameters were measured at baseline, after 3 months with the chokeberry juice supplementation and after the next 3 months without the chokeberry juice supplementation (follow-up period). Significant difference was noticed in the levels of LDL-cholesterol, glycated hemoglobin and serum creatinine (p < 0.05), as well as in the levels of some hematological parameters, such as white blood cell and lymphocyte count (p < 0.01), hematocrit, blood hemoglobin, mean corpuscular volume, hemoglobin and hemoglobin concentration and red blood cell count (p < 0.05). The daily consumption of the chokeberry juice could improve the health status in patients with type 2 diabetes mellitus, in combination with their standard therapy.

Neutrophils play an important role as the central mediators of the innate immune defence response, providing the first line of host protection. It was shown that these cells can trap and kill various microorganisms through different ways. One of them is a release of neutrophil extracellular traps (NETs) composed of chromatin fibrils and antimicrobial proteins. There is the evidence that the release of NETs does not have only a beneficial effect. NETs can trap and kill microorganisms and pathogens, however on the other hand the same pathway can also cause the damage of the organism by various mechanisms. NETs participate in the pathogenesis of a lot of inflammatory and autoimmune disorders, such as thrombosis, atherosclerosis, cystic fibrosis, periodontitis, lupus, rheumatoid arthritis and others. The aim of this review is to summarize information about the release of NETs and their beneficial, but also detrimental effect during various diseases. The better characterization and understanding of the dual role of NETosis during these diseases is necessary for the early diagnosis and more effective treatment.

The present study was designed to examine effects of Sinomenine (SM) on glioma cells growth in vivo and in vitro. Cells growth and apoptosis were detected by MTT assay, TUNEL assay and flow cytometric analysis. In the study, SM treatment led to growth inhibition on a series of glioma cell lines, including U87, U373, U251, Hs683 and T98G. SM prevented U87 growth in the nude mice as well. Inhibitory effects of SM on U87 cells proliferation in vitro and in vivo were more effective than that of temozolomide (TMZ), and SM has synergistic effects with TMZ in the glioma therapy. SM induced apoptotic death in U87 cells via activation of caspase-3, caspase-8 and caspase-9, and down-regulation of HIAP, Bcl-2 and survivin. Moreover, we observed SM decreased the expression of phosphorylated STAT3 (p-STAT3) both in vivo and in vitro. Interestingly, using a specific activator of STAT3, we demonstrated overexpression of p-STAT3 impaired, SM mediated growth inhibition and apoptosis induction in the U87 cells. In summary, our results indicate SM induced growth suppression of human glioma cells through inhibiting phosphorylation of STAT3.

Non-small-cell lung cancer (NSCLC), the most common type of lung cancer, remains the leading cause of cancer death worldwide. Blocking vascular endothelial growth factor (VEGF) signalling is an effective approach to the treatment of NSCLC. Small molecules have been proven to be good resource for discovery of inhibitors of VEGF signalling. DMH4 is a small molecule that we previously developed and demonstrated to have the property of selectively inhibiting VEGF signalling by targeting VEGF receptor 2 (VEGFR2). In this study, we reported that DMH4 can effectively block phosphorylation of VEGFR2 in both H460 and A549 NSCLC cells, which resulted in significant reduction of NSCLC cell viability in a dose-dependent manner, and the growth inhibition (GI50) of DMH4 against H460 and A549 cell lines were 13.27 and 2.75 mm respectively at 24 h. Our further studies demonstrated that DMH4 significantly suppressed migration and invasion of A549 and H460 cells, and induced apoptosis in those cells. Therefore, DMH4 as a small molecular VEGFR2 inhibitor may represent a new valuable drug lead for NSCLC treatment.

Herein, we report a facile route to synthesize palladium nanoparticles (CGPdNPs) using the aqueous fruit extract of C. guianensis Aubl. as a potent biological reducing agent. Reduction of PdCl₂ solution into their nano scale was confirmed with the formation of a black precipitate which gives a reduced absorbance in UV-vis spectroscopy. Fourier transform infrared spectroscopy (FTIR) reveals the active role of phenolic constituents from C. guianensis in reduction and surface functionalization of nanoparticles (NPs). Dynamic light scattering (DLS) and zeta potential analysis confirms the generation of polydispersed highly stable NPs with large negative zeta value (-17.7 mV). Interestingly, X-ray Diffraction (XRD) pattern shows that the synthesized CGPdNPs were face centered cubic crystalline in nature. The HRTEM micrographs of CGPdNPs displays well-dispersed, spherical NPs in the size ranges between 5 and 15 nm with an average of 6 nm. It was also noticed that the synthesized CGPdNPs possess an effective antimicrobial activity against different bacterial pathogens. On the other hand, in vitro cell viability (MTT) assays reveals that the synthesized CGPdNPs exhibited an extraordinary anticancer properties. Eventually, hemocompatibility assay depicts the safe nature of synthesized NPs for biomedical application.

The aim of this study was to compare the effects of DNA repair inhibitors in the context of radio-sensitization of human lung cells. The radio-sensitizing effects of NU7441 (1 mM), an inhibitor of DNA-dependent protein kinase (DNA-PK); KU55933 (10 μM), an inhibitor of ataxia-telangiectasia mutated kinase (ATM); and VE-821 (10 μM), an inhibitor of ATM-related kinase (ATR) were tested by the xCELLigence system for monitoring proliferation, fluorescence microscopy for DNA damage detection, flow-cytometry for cell cycle and apoptosis analysis and western blotting and ELISA for determination of DNA repair proteins. We employed normal human lung fibroblasts (NHLF, p53-wild-type) and non-small cell lung cancer cells (H1299, p53-negative). DNA-PK inhibition (by NU7441) in combination with ionizing radiation (IR) increased the number of double strand breaks (DSB), which persisted 72 h after irradiation in both cell lines. Additionally, NU7441 and KU55933 in combination with IR caused G2-arrest. ATR inhibitor (VE-821) together with IR markedly inhibited proliferation and induced G2/M arrest accompanied by apoptosis in H1299, but not in NHLF cells, and thus diminished DNA-repair of tumour cells but not normal lung fibroblasts. Our findings indicate that ATR inhibition could be a promising therapeutic strategy in p53-deficient lung tumours.

Silver nanoparticles (AgNPs) with aqueous leaf-extract of the timber-yielding plant Anogeissus acuminata were synthesized for in vitro control of pathogenic bacteria. Characterization of AgNPs with ultraviolet-visible spectroscopy, scanning electron microscopy (SEM), energy dispersive X-ray (EDX) spectroscopy, X-Ray diffraction (XRD) study and Fourier transformed infrared spectroscopy (FTIR) study was done for a confirmation of the synthesis. The SEM analysis confirmed that the metal particles were below 100 nm size. The antibacterial activity of AgNPs was monitored by agar-well diffusion method against 11 multidrug resistant (MDR) urinary tract infection (UTI) causing pathogenic bacteria, isolated from clinical samples. At 15 μg/ml AgNPs, values of the zone of inhibition (ZI) ranged from 19 to 13 mm, while against the standard antibiotic, gentamicin 30 μg/ml ZI ranged from 28 to 20 mm. Host toxicity testing of AgNPs with cultured lymphocytes from human umbilical cord blood in vitro was done; at 3000 mg/l AgNPs, 25% of cell death occurred. Thus, the synthesized AgNPs with aqueous leaf extract of A. acuminata could control most MDR UTI bacteria without any toxicity to human lymphocytes.

Ischemic stroke is a severe cause of disability and death all over the world. To search for effective therapy for ischemic stroke, PC12 cells damaged by oxygenation and glucose deprivation/restoration were employed to assess the protective effects of inotodiol. As a result, inotodiol can improve the cell viability and attenuate the leakage of lactate dehydrogenase. Meanwhile, inotodiol can prevent oxidative stress by reducing reactive oxygen species generation, decreasing the content of malonic dialdehyde, and increasing the activity of superoxide dismutase. In addition, the dysfunction of mitochondria induced by oxygenation and glucose deprivation/restoration was ameliorated through decreasing the level of intracellular calcium and increasing the mitochondrial membrane potential. At the same time, inotodiol can inhibit PC12 cells apoptosis through downregulation of Caspase-3 and Bax as well as upregulation of Bcl-2. These results reveal inotodiol can protect PC12 cells against the injury induced by oxygenation and glucose deprivation/restoration. This investigation gives promising evidences for the therapy of ischemic stroke.

The urokinase-type plasminogen activator (uPA) and PA inhibitor 1 (PAI-1) play important roles in breast cancer metastasis through cell migration and invasion. They are clinically applicable prognostic and predictive markers. High levels of uPA and PAI-1 are associated with high risk of recurrence and adjuvant chemotherapy provides substantial benefit for this breast cancer population. The current sole validated method for quantifying uPA level in breast tumour tissue is ELISA assay. It requires 50-300 mg of fresh or frozen tissue, which is the main limitation for routine use. In this study, we evaluated the performances of customized antibody microarray to quantify uPA concentration from reduced extraction solution of breast tumour tissue and compared it with standard ELISA kit. We firstly optimized the elaboration of customized antibody microarray in order to sensitively detect and quantify uPA standard solutions. In the best conditions, we analysed uPA concentration in 16 cytosolic extracts from breast tumour tissue. Results showed that our customized antibody microarray could correctly quantify uPA concentration while consuming 100 times less volume of tumour tissue extraction solution than ELISA. Our antibody microarray is a powerful and promising tool for the miniaturization of the immunoassay quantification of uPA from breast tumour tissue extracts.

Pseuduvaria macrophylla (Family: Annonaceae) is commonly used as medicinal plant in Malaysia. A recent study with the Pseuduvaria species showed antioxidant and antidiabetic effects. This study aimed to ascertain antidiabetic potential of methanolic extract of Pseuduvaria macrophylla bark (PM) using streptozotocin-nicotinamide induced diabetic rat models. Various phytochemical and biochemical properties of the plant have been evaluated. The results showed that the extract has potentially normalized the elevated blood glucose levels by upregulating the insulin and C-peptide levels and alleviated oxidative stress by improving glutathione (GSH) and reducing lipid peroxidation (LPO) in the diabetic rats. In addition, PM has drastically downregulated the levels of pro-inflammatory cytokines and transforming growth factor beta-1 (TGF-β1). Histopathological examination of the pancreas in PM treated diabetic rats showed significant recovery of the pancreatic structural degeneration and thus reflected the protective role of PM against peroxidation damage by a rise in insulin level as evidenced by the immunohistochemistry study. The improved expressions of GLUT-1, GLUT-2 and GLUT-4 further confirmed the restoration of β-cell mass by PM. Interestingly, the findings demonstrated the antioxidant, anti-inflammatory and antihyperglycemic potential of PM which may provide future lead for the management of type-2 diabetes.

As a common complication of diabetes mellitus (DM), diabetic cardiomyopathy (DCM) is considered to be one of the major causes of mortality and morbidity. The therapeutic effects of naringenin have been verified in the treatment of various human diseases. However, the application of naringenin in the treatment of DCM still has not been reported. In this study, human AC16 cardiac cells were treated with normal d-glucose and high d-glucose (HG). After transfection with miR-30d-5p inhibitor, Cell Counting Kit-8 (CCK-8) method was used to measure cell viability. Hoechst 33258 staining was performed to observe the morphological changes of nucleus. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the activity of caspase-3. Cell apoptosis was detected by Annexin V-FITC/propidium iodide (PI) staining. Levels of light chain 3 (LC3) including LC3-I and LC3-II as well as nucleoporin p62 (P62) were detected by Western blot. We found that Naringenin treatment increased the reduced cell variability caused by HG treatment. Naringenin also increased expression level of miR-30d-5p in human AC16 cardiac cells after HG treatment. Treatment with miR-30d-5p inhibitor reduced the effect of miR-30d-5p in increasing cell variability and reducing cell apoptosis. Naringenin treatment reduced the increased levels of LC-I, LC-II and P62, but miR-30d-5p inhibitor reduced those changes. Therefore we concluded that naringenin could alleviate HG-induced injuries through the upregulation of microRNA-30d-5p level in human AC16 cardiac cells.

The effect of moderate and overtraining exercise on Th1/Th2 balance was evaluated in rat splenocytes. Male Wistar rats were divided into sedentary control (C), moderately trained (MT; V = 20 m/min, 30 min/day, 8 weeks), overtrained (OT; V = 25 m/min, 60 min/day, 11 weeks) and recovered after overtraining (OR) (OT plus 2 weeks recovery) groups. At the end of study, cell viability, proliferation, interleukin 4 (IL-4) and interferon-γ (IFN-γ) secretion were evaluated in non-stimulated, phytohemagglutinin (PHA) and concavaline A (Con A)-stimulated splenocytes. Cell viability increased in MT and OR groups compared to control. Cell proliferation was higher in OR group than other groups. IL-4 concentration in PHA-stimulated cells from MT and OT groups, and IL-4 concentration in Con A-stimulated cells from OR and OT groups, were higher than the control group, but not for IFN-γ. In non-stimulated cells, IFN-γ/ IL-4 ratio was higher than MT and OT groups. In PHA and Con A-stimulated cells, IFN-γ/ IL-4 ratio was lower in exercise groups than control. We previously showed that moderate exercise increases Th1 cytokines in serum, but in splenocytes, Th2 or Th1 response may increase depending on the type of mitogen stimulation. Two-week recovery restored Th1/Th2 balance, only in non-stimulated splenocytes of overtrained animals.

This study was designed to find the correlation between changes in CD⁴⁺ cell count, lipid profile and liver marker enzymes in HIV-infected and AIDS patients. The study population consisted of 150 subjects, age and sex-matched and divided into three groups [control subjects (n=50), HIV infected (n=50) and AIDS patients (n=50)]. We observed a significant reduction in CD⁴⁺ cell count in HIV/AIDS patients when compared to control subjects. Serum levels of total cholesterol, high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were found to be decreased significantly in HIV/AIDS patients when compared with normal counterparts. On the other hand, the levels of triglyceride (TG) and very low-density lipoprotein cholesterol (VLDL-C) were markedly elevated in HIV/AIDS patients compared to normal subjects. The activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) observed in HIV/AIDS patients were significantly higher than in the control group. Further, the above mentioned haematological and biochemical variables were found to be affected more significantly in AIDS patients when compared with HIV infected subjects. Hence, it may be concluded that CD⁴⁺ cell count, lipid profile and liver enzymes can be a good index of disease progression in HIV infection and AIDS patients.

Zingiber officinale Roscoe, Ginger, has been used in folk medicine as a medicinal plant, as well as a spice and food in many countries. This research was carried out to evaluate the antiparasitic effect of ginger root extract (GE) and GE/F1 (fraction 1 obtained from GE) against Toxoplasma gondii (T. gondii) in vitro and in vivo. The effects of GE and GE/F1 against the proliferation of T. gondii-infected C6 cells and T. gondii were evaluated by several indicators such as an MTT assay, nuclear staining, immunofluorescence staining, apoptotic proteins and animal testing. GE/F1 strongly inhibited the proliferation of T. gondii-infected C6 cells and T. gondii in a dose-dependent manner compared with sulfadiazine. After T. gondii invasion, C6 cells induced the activation of caspase-3, bax, p53 and p21 related to programmed cell death, and GE/F1 effectively suppressed the expression of caspase-3, bax, p53 and p21 causing cell death of the infected host cells. In addition, INF-γ, and IL-8 levels, and the viability of T. gondii-infected mice treated with GE/F1 (500 μg/ml) were not changed or increased during the period of the experiment. These results demonstrate that GE/F1 not only induces anti-T. gondii effects causing the inactivation of apoptotic proteins in infected host cells through the direct inhibition of T. gondii but also has antiparasitic properties which inhibit inflammatory cytokine secretion in vivo.

To evaluate the in vitro effects of sunitinib malate and meloxicam in isolation, and to analyse the ability of meloxicam to enhance the cytotoxicity of sunitinib malate in three human bladder-cancer cell lines. Cell lines were treated with sunitinib malate and meloxicam, either in isolation or combined. Leishman staining, MTT method, comet assay, MDC staining and M30 CytoDEATH antibody were performed. The Chou and Talalay method was applied. Sunitinib malate and meloxicam supressed cell proliferation in bladder-cancer cells in isolation, in a concentration-dependent manner. Treatment of bladder-cancer cells with a combination of sunitinib malate and meloxicam showed a synergistic effect. When exploring the mechanism of this combination by means of comet assay, there is the suggestion that meloxicam increases sunitinib malate cytotoxicity through DNA damage. Autophagic and apoptotic studies show a greater incidence of autophagic vacuoles and early apoptotic cells when the combined treatment was put into use. In isolation, sunitinib malate and meloxicam demonstrated anti-tumour effects in our study. Furthermore, simultaneous exposure of cells to sunitinib malate and meloxicam provided a combinatorial beneficial effect. This hints at the possibility of a new combined therapeutic regimen, which could lead to improvements in the treatment of patients with bladder cancer.

β-catenin signaling is required for hair follicle development and regeneration which are involved in the resuscitation of hair follicle stem cells (HFSCs). To further characterize the role of β-catenin in the regulation of proliferation of HFSCs, the β-catenin expression was measured in the defined stages of hair follicle cycle and the proliferative potency was determined by using an in vitro cell growth assay. Our results showed that activation of β-catenin correlated with HFSCs proliferation, which appeared to be mediated by the nuclear translocation of stabilized β-catenin and the activation of responsible cell cycle genes (cyclin D1 and p21). In addition, PI3K/Akt pathway was also involved in the HFSCs proliferation, partly regulated by β-catenin signaling pathway. These results demonstrate that β-catenin is an essential factor in the regulation of HFSCs proliferation via PI3K/Akt pathway and might be a potential therapeutic target for the regulation of the yield of keratinocytes from HFSCs.

In this study we analyzed molecular mechanisms of antioxidative protection of thymol and thyme oil using differentiated PC12 cells, a widely accepted neuronal model. Thymol due to multiple functions is commonly used for clinical applications. However, its action on nervous tissue remains poorly understood. We evaluated the effect of 24 h incubation of the cells with thymol (100 and 400 μM) and equivalent content of thyme oil. Microsomal glutathione transferase 1 (Mgst1) is an important player in anti-oxidative protection because of its transferase and peroxidase activities. Since its expression decreases during aging, we also used stable transfected cells with downregulated Mgst1 (PC12_M). We analyzed cell viability, lipid peroxidation level, glutathione content and expression of key enzymes responsible for cellular reduced glutathione - catalytic subunit of γ-glutamylcysteine ligase and glutathione reductase. Whereas thymol and thyme oil improved antioxidant capacity of control cells, diminished protection was observed in PC12_M line. Increasing interest in natural dietary components has focused attention on plants used as a rich source of bioactive phytochemicals. However, currently available information on the safe amount for thyme oil using appears to be insufficient. Our results indicate that the thyme oil should be used with caution, especially by elderly people.

Diethyldithiocarbamate (ditiocarb), a metabolite of the old anti-alcoholic drug disulfiram (Antabuse), forms proteasome-inhibiting metal complexes with copper or zinc that suppress cancer cells both in vitro and in vivo. The drug has been used in a clinical trial (NCT00742911) along with copper gluconate as a dietary supplement in patients with cancer spreading to the liver. In this study, we demonstrate the effect of synthetic complexes of disulfiram with four various metals (Mn, Fe, Cr and Cu) used as food supplements. These complexes may be spontaneously formed in the blood during the use of disulfiram with divalent metals and thus may suppress the growth of cancer in vivo. The cytotoxic effect of the compounds and the compounds' ability to inhibit the cellular proteasome were tested in the osteosarcoma cell line U2OS. After 48 h, copper and manganese complexes exhibited cytotoxic effect on the cell line, in sharp contrast to both iron and chromium complexes.