Iranian Journal of Pharmaceutical Research
Volume:18 Issue: 2, Spring 2019

Sustained release of thermal-instable and water-soluble drugs with low molecule weight is a challenge. In this study, sodium bicarbonate was encapsulated in ethyl cellulose microspheres by a novel solid-in-oil-in-oil (S/O/O) emulsification method using acetonitrile/soybean oil as new solvent pairs. Properties of the microspheres such as size, recovery rate, morphology, drug content, and drug release behavior were evaluated to investigate the suitable preparation techniques. In the case of that the ratio of the internal and external oil phase was 1: 9, Tween 80 as a stabilizer resulted in the highest drug content (2.68%) and a good spherical shape of microspheres. After the ratio increased to 1: 4, the microspheres using Tween 80 as the stabilizer also had high drug content (1.96%) and exhibited a sustained release behavior, with 70% of drug released within 12 h and a sustained release of more than 40 h. Otherwise, different emulsification temperatures at which acetonitrile was evaporated could influence the drug release behaviour of microspheres obtained. This novel method is a potential and effective method to achieve the encapsulation and the sustained release of thermal-instable and water-soluble drugs with low molecule weight.

Novel biodegradable Poly (L-lactic acid) (PLLA) microspheres containing ytterbium were designed for intra-tumoral radiotherapy, especially for radioembolization. 175Yb possess both therapeutic beta and diagnostic gamma radiations. In this work, process to make ready radiomicrospheres 175Yb(acac)3-loaded PLLA for more consideration has been investigated. The radiomicrospheres were prepared with approximate size of 20-40 µm, and radionuclidic purity > 92%. The radiomicrospheres were stable in vitro for up to 72 h in normal saline, and also in human serum albumin (HSA). Biodistribution in mice bearing 4T1 tumor showed specific radionuclide uptake over 48 h. Tumor necrosis was also observed at the injection site up to 12 days after injection. These data showed that 175Yb-PLLA microspheres could be prepared and considered further for radiomicrospheres tumor therapy.

In this study a solid dispersion and a physical mixture of cilazapril (CIL) with a biopolymer - polyvinylpyrrolidone (PVP) as a carrier were prepared so as to investigate the effect of PVP on the stability of CIL. CIL is unstable in solid state and decomposes rapidly under humid conditions. It requires stabilization to ensure safety of its use. The studied CIL/PVP formulations were prepared by milling and evaporation technique. Their identity was confirmed by FT-IR method. The stability of CIL in the CIL/PVP formulations was assessed by forced ageing test under isothermic conditions using RP-HPLC. The influence of temperature (experimental conditions: RH 76.4% and T = 70, 75, 80, 85, and 90 oC) and the effect of relative humidity (experimental conditions: RH 25.0%, 50.9%, 60.9%, 66.5%, 76.4%, T = 90 °C) on the rate of CIL degradation were examined. It was established that the process of CIL decay in the studied forms followed first-order kinetics with the formation of one degradation product - cilazaprilat. The degradation rate constant of this reaction was lower than that for pure CIL. The energy of activation of the CIL degradation in the presence of PVP was higher than that of pure CIL. Furthermore, CIL incorporated into PVP exhibited lower sensitivity to moisture. Based on these data PVP was considered as a potential stabilizing substance for CIL-containing dosage forms.

Fifteen novel β-lactams bearing the N-ethyl tert-butyl carbamate group 5a-o and fifteen N-(2-aminoethyl) β-lactams 6a-o were synthesized by the ketene-imine [2+2] cycloaddition reaction (Staudinger ). The cycloaddition reaction was found to be totally diastereoselective leading exclusively to the formation of the cis-β-lactam derivatives. These newly synthesized β-lactams were evaluated for their antimalarial activity against p. falciparum K14 resistant strain and showed good to excellent EC50 values. Of the thirty β-lactams tested, 5h, 6a and 6c showed IC50 < 20 µM while 5b, 5c, 5e, 5f, 5g, 5i, 5j, 6d, 6g and 6h exhibited IC50

New quinazoline derivatives were prepared by one pot reaction of anthranilic acid, acetic anhydride and primary amines, under ultrasonic irradiation. As a result, Ultrasonic irradiation has led to affordable, clean synthesis of a variety of target compounds in much higher yields, than traditional methods. This method has numerous advantages: such as higher yields, shorter reactions time, and easier work-up. Several structural classes among these compounds were identified to have vasorelaxant activity. In this respect, all of the newly synthesized quinazolinone derivatives displayed vasorelaxant properties on the isolated thoracic rat aorta. The IC50 of compounds 2a (-6.00 ± 0.55), 2g (-7.31 ± 0.94), 2n (-7.15 ± 0.81) and 2p (-7.77 ± 0.31) was comparable to that seen in the Acetylcholine (-7.13 ± 0.14). The structures of the newly synthesized compounds were confirmed by IR, 1H NMR, 13C NMR, mass spectral studies, elemental analysis, and melting point.

A simple and efficient homogenous liquid-liquid extraction method performed in a narrow tube combined with dispersive liquid-liquid microextraction method has been presented for the simultaneous determination of two antiepileptic drugs in urine followed by gas chromatography with flame ionization detection. In this method, a mixture of acetonitrile and urine sample (homogenous solution) is loaded into a column partially filled with solid sodium chloride. By passing the homogenous solution through the salt layer, acetonitrile is separated from the aqueous solution as the fine droplets and collected on top of the column as a separated phase. The obtained organic phase is removed and mixed with an extraction solvent, and then the resulting mixture is rapidly injected into an alkaline solution. Various experimental parameters affecting performance of the proposed method such as type and volume of extraction solvent, pH, and flow rate in homogenous liquid-liquid extraction step, and type and volume of extraction solvent and ionic strength in dispersive liquid-liquid microextraction step were investigated. The relative standard deviation of the proposed method was <8% (n = 6, C = 1 µg L-1 of each analyte). The limits of detection for phenobarbital and carbamazepine were 0.017 and 0.010 µg L-1 and the limits of quantification were 0.056 and 0.033 µg mL-1, respectively.

Iguratimod is a new synthetic disease-modifying antirheumatic drug intended to treat patients with rheumatoid arthritis. A new method using recombinant human CYP450s yeast cells containing c-DNA expressed P450s was applied to identify the metabolic pathways of iguratimod and to prepare its metabolite. The metabolite was isolated, and its structure was identified by quadrupole time-of-flight-mass spectrometry and nuclear magnetic resonance. Furthermore, a selective and sensitive high performance liquid chromatography (HPLC) method was developed for the simultaneous quantification of iguratimod and its major metabolite in rat plasma for the first time. The results indicated that iguratimod was mainly metabolized to a metabolite by CYP2C9 and CYP2C19 in in-vitro study. The structure of the metabolite was identified as M2 (N-[3-(acetamido)-4-oxo-6-phenoxy-4H-chromen-7-yl]methanesulfonamide). HPLC assay was achieved on a C18 column using methanol-water containing 0.1% trifluoroacetic acid (55:45 v/v) at a flow rate of 1 mL/min with UV detection at 257 nm. Standard calibration curves were obtained in the concentration range of 0.5–20 µg/mL for iguratimod and its metabolite M2. The lower limits of detection of iguratimod and M2 in rat plasma were 0.1 and 0.25 µg/mL, respectively. The intra- and inter-day precision (RSD%) were within 5% for the two analytes. The average recoveries of the analytes were greater than 90%. In conclusion, recombinant human CYP450s whole-yeast transformation system could be successfully used to identify and prepare the major metabolite of iguratimod. The HPLC method we developed could be successfully applied to evaluate pharmacokinetics of iguratimod and its metabolite M2 in rats.

In the present study, a reusable graphene oxide (GO) based dispersive-solid phase extraction (d-SPE) was synthesized and applied for the analysis of trace ampicillin sodium (AMP) and clindamycin hydrochloride (CLI) in water samples followed by high performance liquid chromatography-UV detection (HPLC-UV). Batch experiments were conducted to investigate the effects of pH and volume of the sample solution, contact time, adsorption isotherms, temperature, and desorption conditions. The maximum adsorption capacities of AMP and CLI on GO nanosheets were found to be 33.33 mg g-1 and 47 mg g-1, respectively. The adsorption isotherm data can be well fitted by Temkin (AMP and CLI) and Freundlich (AMP), and the kinetics of adsorption followed the pseudo-second-order rate equation. The thermodynamic parameters were calculated, indicating that the adsorption process of both analytes were spontaneous and exothermic. In addition, the d-SPE following HPLC analyses showed good linearity in the range of 0.5-200 ng mL-1 (R2= 0.999) for AMP and 1-200 ng mL-1 (R2= 0.999) for CLI, with LOD of 0.04 and 0.24 ng mL-1 for AMP and CLI, respectively. The percent of extraction recoveries and intra and inter-day precisions (expressed as RSD %, n=3) were in the range of 96.4-101.6%, 2.2-3.0 and 3.7-4.7 for AMP and 94.2-98.6%, 2.2-3.8 and 3.5-4.6 for CLI, respectively. The preconcentration factor of 20 was achieved for both analytes. According to these results, it can be concluded that the validated technique is a simple, cost-effective and repeatable for analysis of AMP and CLI in water samples.

Pharmaceutical drugs play an important role in human life since they caused a revolution in human health. Notably, their administration to a living organism helps body to stay healthy. Commonly, they are employed to diagnose, prevent, or treat and cure a disease via a biological effect on a human body. Administration of impurity-free and adequate amounts of pharmaceutical compounds would be beneficial. Therefore, the quantity and purity of the substances in pharmaceutical compounds are continuously monitored during drug manufacturing with various chemical or instrumental analysis techniques. The possibility of impurities development and chemical or quantity changes of active drug species at various stages (namely during production, transportation or storage) makes them redundant and risky for human health. So, sensitive and accurate analysis methods for qualitative and quantitative analysis are highly demanded by pharmaceutical companies and medical centers. The present mini review emphasizes on application of carbon based modified electrodes in health care and pharmaceutical analysis. Electrochemical determination of drugs employing carbon nanostructured modified electrodes will be reviewed and their advantages and disadvantages will be mentioned.

AIn this study a simple and reliable stability-indicating RP-HPLC method was developed and validated for analysis of Dexpanthenol in an artificial tear formulation. The chromatographic separation was performed on a HPLC C18 column (25.0 cm  4.6 mm, 5 m) using a mixture of 0.037 M monobasic potassium phosphate in water (adjusted with 0.1% (v/v) phosphoric acid to a pH of 3.2) and methanol (90:10). The flow rate was set at 1.5 ml/min and Dexpanthenol concentration was determined at λmax = 205 nm. The HPLC analysis method was validated in the terms of linearity, precision, accuracy, specificity, sensitivity according to International Conference on Harmonization (ICH) guidelines. The results showed that the retention time was 8 min and no interferences were observed from the formulation excipients and stress degradation products. Linear regression analysis of data for the calibration plot showed a linear relationship between peak area and concentration over the range of 10 - 100 μg/ml; the regression coefficient was 0.996 and the linear regression equation was y = 20.011 x + 146.83. This HPLC method was precise and accurate in the range of 10 – 100 μg/ml. Also the dexpanthenol concentration in artificial tear formulation was determined by this HPLC method, which was in accordance with the label claimed. This validated HPLC method could be used for routine analysis, quality control and the stability of analysis of eye gel containing dexpanthenol formulations.

In present study, a DLLME-HPLC-UV method was developed and validated for the extraction, pre–concentration, and subsequently quantification of vitamin D3 (Vit D3) in milk and yogurt samples. In order to be able to extract Vit D3 from studied samples efficiently, the DLLME procedure was optimized with respect to the parameters affecting the extraction efficacy, where acetonitrile (2 mL as disperser solvent) resulting from the protein precipitation procedure was mixed with 80 µL carbon tetrachloride (as an extraction solvent) respectively. The extracted samples were quantitatively analyzed with a HPLC technique using a C8 column (250 mm × 4.6 mm, 5 μm) at room temperature (25 °C), mobile phase of acetonitrile/methanol (90:10% v/v) in isocratic elution mode at a flow rate of 1.2 mL/min and UV detection at 265 nm. The method validation results revealed that the method was linear in the concentration range of 2 to 60 ng/mL (r = 0.9997) with a LOD of 0.9 ng/mL and LLOQ of 2 ng/mL; the method was accurate (-2.1% ≤ RE% ≤ +0.6%) and precise (1.2% ≤RSD% ≤ 11.3%) and its recovery was in the range of 86.6 to 113.3%. The obtained results indicated that the method could be utilized as an easy to use technique for the monitoring Vit D3 in dairy products, especially milk and yogurt samples.

Cell-based biosensors (CBBs) are becoming important tools for biosecurity applications and rapid diagnostics in food microbiology for their unique capability of detecting hazardous materials. Pollutants, such as heavy metals and chemicals, are now considered as a global threat and are associated with detrimental health outcomes. Fast and accurate detection of pollutants is essential to reduce these threats. In this study, the enhancer sequence of human cytomegalovirus (hCMV) IE genes cloned upstream of luc gene in PGL4.26 plasmid, in order to increase basal luciferase activity. This recombinant vector was transfected into Huh7 cell line and after 21 days of treatment with Hygromycin B selectable marker, stable cell line was generated. After several passages, cells containing this vector showed high luciferase activity in normal conditions without any induction following to overexpression of luc gene. Huh7-CMV-luc cell line was able to detect the slightest changes in ATP level, due to the effect of different toxins on the cell which disrupt cellular respiration and ATP production processes. In order to investigate the sensitivity of the cell line, cells were incubated with 0.1-10 μM of chemical toxins affecting ATP production. This toxins affect luciferase activity in a dose dependent manner, with maximal sensitivity approximately about 0.2 μM to toxin concentrations. Additionally, this biosensor provided a rapid detection as early as 4 h in response to the toxicants. Whole cell biosensors like huh7-CMV-luc cell line can be considered as a powerful tool for the sensitive and efficient monitoring of general toxins, drugs and environmental pollutants.

All sophisticated methods for direct determination of methanol require advanced instruments and high technical knowledge which preparing them is very expensive in none developed and developing countries. This work reports a simple and efficient qualitative technique for semi determination of methanol content in herbal distillates by a new modified chromotropic acid method. The technique is based on the indirect detection of methanol after its oxidation and transforming to formaldehyde by chromotropic acid (formaldehyde specific color indicator). To measure methanol level in herbal distillates, a water diluted sample were mixed with 50µl of sulfuric acid and potassium permanganate and after 5 minutes, followed by addition sodium bi-sulfite, chromotropic acid and concentrated sulfuric acid in two separated steps and finally, eye comparing with four color standard tubes which gives a range of amount of methanol in the sample. The method has a good precision and accuracy and its Limit of Detection is 25 mgL-1. It is particularly suitable for semi quantitative measurement of methanol in herbal distillates not only in the production process quality control of workshops or small companies with no laboratory equipment and adequate financial properties but also the quality check of point of-sale samples from commercial markets. To the best of our knowledge, there isn’t any report about such method.

Acetaminophen (APAP) toxicity threatens human health due to increased mortality associated with its overdose. Doxycycline (DC) because of its properties such as antioxidant and anti-inflammatory can be a good therapeutic strategy to treat the acute toxicity induced by APAP. Male mice were divided to six groups in two periods of 3 and 24-h as normal saline, APAP 400 mg/kg, DC 100 mg/kg and groups treated by 25, 50 and 100 mg/kg DC just before APAP, respectively. At the end of the 3-h and 24-hour periods, the hepatic index, biochemical parameters including serum aspartate transaminase (AST) and alanine transaminase (ALT) activity and hepatic catalase activity, reduced glutathione (GSH) and malondialdehyde (MDA) levels in liver and histopathological changes were evaluated. The results indicated that DC had no apparent effect on the hepatic index but significantly normalized the level of biochemical parameters and reduced APAP induced liver damage. Overall, it be concluded that DC can inhibit or resolve harmful effects of APAP through antioxidant and anti-inflammatory properties. However, more studies are needed to understand exact mechanism of DC and its application for clinical use.

The TiO2, which is a main material in the field of photocatalytic reactions, includes rutile and anatase phase. Titanium dioxide has possessed notice due to its promising applications in the environmental photocatalytic degradation of pollutants of organic compound in waste water and utilization of solar energy. The nanosized manganese titanate (pyrophanite) MnTiO3 was collected by oxidation of Mn(OH)2 with TiO2 powder in cetyltrimethylammonium bromide (CTAB) micelle solutions and the calcinations of the produced powders. Therefore, it was decided to determine the Mechanistic mitochondria toxicity of nanoparticles towards liver, kidney, heart, and brain via new and reliable methods. Our results showed that nanoparticles induced mitochondria dysfunction via an increase in ROS production and membrane potential collapse, correlated to cytochrome c release. Also, increased disturbance in oxidative phosphorylation was also shown by the decrease in ATP. Recent studies have suggested that nanoparticles leading to cytosolic release of lysosomal content, and ultimately apoptosis. This study suggests that mitochondrial oxidative stress and impairment of oxidative phosphorylation in vital organ Mitochondria may play a key role in manganese titanate toxicity.

Scorpion venom is a rich source of toxins which have great potential to develop new therapeutic agents. Scorpion chloride channel toxins (ClTxs), such as Chlorotoxin selectively inhibit human Matrix Methaloproteinase-2 (hMMP-2). The inhibitors of hMMP-2 have potential use in cancer therapy. Three new ClTxs, meuCl14, meuCl15 and meuCl16, derived from the venom transcriptome of Iranian scorpion, M. eupeus (Buthidea family), show high sequence identity (71.4%) with Chlorotoxin. Here, 3-D homology model of new ClTxs were constructed. The models were optimized by Molecular Dynamics simulation based on MDFF (molecular dynamics flexible fitting) method. new ClTxs indicate the presence of CSαβ folding of other scorpion toxins. A docking followed by steered molecular dynamics (SMD) simulations to investigate the interactions of meuCl14, meuCl15 and meuCl16 with hMMP-2 was applied. The current study creates a correlation between the unbinding force and the inhibition activities of meuCl14, meuCl15 and meuCl16 to shed some insights as to which toxin may be used as a drug deliverer. To this aim, SMD simulations using Constant Force Pulling method were carried out. The SMD provided useful details related to the changes of electrostatic, van de Waals (vdW), and hydrogen-bonding (H-bonding) interactions between ligands and receptor during the pathway of unbinding. According to SMD results, the interaction of hMMP-2 with meuCl14 is more stable. In addition, Arginine residue was found to contribute significantly in interaction of ClTxs with hMMP-2. All in all, the present study is a dynamical approach whose results are capable of being implemented in structure-based drug design.

One of the emerging therapeutic strategies for targeted therapy of cancer is the use of chimeric proteins. The p28 peptide has the ability of selective entrance and activating apoptosis in breast cancer cells. The NRC antimicrobial peptide showed cytotoxic activity on various breast cancer including drug-resistant variants but also normal cell lines. Here we designed a chimeric protein consisted of these peptides to determine their targeted and enhance cytotoxic effects on breast cancer cells. The chimeric protein was cytotoxic to MDA-MB-231 and MCF7 breast cancer cell lines in a dose-dependent manner after 48 h of treatment. In addition, the cytotoxic effects of the p28 alone were significantly lower than the chimeric protein indicating the additive or enhanced effects of the two peptides. Flow cytometry analysis showed that the induced cell death is mediated via apoptosis. The designed chimeric protein had enhanced effects on breast cancer cell lines and exerts its anticancer effects on MCF7 breast cancer cells through mitochondrial caspase dependent and -independent apoptotic pathways. Taken together, the results of this study suggested the chimeric protein to be a reasonable anti-cancer agent which must be further evaluated by subsequent in-vitro and in-vivo preclinical studies.

Methamphetamine (METH) abuse causes neurodegeneration. Medicinal herb such as crocin has neuroprotective properties. The current study evaluates the role of CREB-BDNF signaling pathway in mediating the neuroprotective effects of crocin against METH-induced neurodegeneration in rats. Sixty adult male rats were divided randomly into group 1 and group 2 which received 0.7 mL/rat of normal saline and 10 mg/kg of METH intraperitoneally (i.p) respectively, and groups 3, 4, 5 and 6 which treated concurrently with METH (10 mg/kg) and crocin(10, 20, 40 and 80 mg/kg I.P respectively) for 21 days. Morris water maze(MWM)was used to evaluate cognitive activity. According to the critical role of hippocampus in cognitive behavior, the molecular and biochemical parts of our study were done in hippocampus and according to this, hippocampal neurodegenerative parameters and also CREB and BDNF levels were evaluated in isolated hippocampus. METH disturbed the learning, memory, and simultaneous treatment with various doses of crocin reduced the METH-induced cognition disturbances. In addition, METH treatment increased lipid peroxidation and the levels of oxidized form of glutathione (GSSG), interleukin 1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), and Bax, while reducing reduced form of glutathione (GSH), Bcl-2, P-CREB, and BDNF levels in the hippocampus. METH also reduced the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) in the hippocampus. In contrast, crocin (40 and 80 mg/kg) attenuated METH-induced apoptosis, oxidative stress, and inflammation, while elevating P-CREB and BDNF levels. Thus, crocin confers neuroprotection against METH-induced neurodegeneration in hippocampus and this is probably through activation of P-CREB/BDNF signaling pathway.

Tumor necrosis factor alpha (TNF-α) is an inflammatory cytokine which plays crucial roles in pathogenesis of inflammatory diseases. The current study aimed to investigate the binding abilities of I44 and I49 domain antibodies to TNF-α. The dAbs were expressed in bacterial expression system and purified by affinity chromatography using Ni-sepharose column. The expression and purity of the proteins were evaluated using western blotting and SDS-PAGE techniques, respectively. ELISA experiment showed that I44 and I49 dAbs bind to TNF-α with the binding constants (Kd) of 5.18 ± 1.41 and 2.42 ± 0.55 µM, respectively. The inhibitory effect of dAbs on TNF-α biological effect was determined in MTT assay in which I44 and I49 prevented TNF-α cell cytotoxicity with IC50 values of 6.61 and 3.64 µM, respectively. The identified anti-TNF-α dAbs could bind to and inhibit TNF-α activity. The dAbs activities can be attributed to their ability to establish hydrogen bonds as well as hydrophobic contacts with TNF-α. The results of the current study can pave the way for further structural studies in order to introduce new more potent anti-TNF-α antibodies.

Severe physical activity leads to a sharp increase in free radicals, an oxidative stress, inflammation, and tissue damage. Resveratrol as one of the antioxidants can be effective in preventing the effects of oxidative stress. Therefore, the present study was aimed to evaluate the effect of trans-resveratrol supplementation and training exercise on inflammation-related factors. Sixty-four male Wistar rats were divided into six groups, each group consisting of 16 animals: 1) excursive + trans-resveratrol, 2) exercise group, 3) trans-resveratrol group, and 4) control group. Following the familiarizationsessions, a more consistent protocol with an intensity of 65% vo2 max was performed for 12 weeks. Afterward, half of the mice in each group received acute exercise training with an intensity of 70-75% of vo2 max at the age of 20 weeks, until reaching the disability level. Finally, the levels of inflammatory markers were measured using special kits. Our findings depicted that inflammatory factors such as CPR, TNF-α, IL-6, and IL-7 were not affected by endurance protocol (P > 0.05), whereas, they were significantly increased by acute exercise training (P > 0.05). Additionally, we found that RES supplements led to a decrease in CPR and IL-6 levels, while not affecting TNF-α and IL-17 levels. According to available evidence, RES appears to have anti-inflammatory and protective effects during exercise by reducing inflammatory factors. Further studies are required to clarify the role of trans-resveratrol supplementation after exercise training.

Chronic morphine (CM) treatment increases the phosphorylation of the mammalian target of rapamycin (mTOR), which confers neuroprotection against ischemia/reperfusion (I/R) injury. Besides its important regulatory role in the proliferation, metabolism, and survival of cells, the mTOR is critically involved in intracellular signaling events during I/R injury. In the present study, we investigated the interaction between the expressions of the mTOR and inducible nitric oxide synthase (iNOS) and their possible protective effects on hippocampal neurons against I/R injury in morphine-dependent mice. Additive doses of morphine were administered for 5 days to BALB/c mice so as to induce CM preconditioning before I/R injury. Global brain ischemia was induced via the occlusion of bilateral common carotid arteries for 30 minutes. CM attenuated iNOS expression, NO production, and malondialdehyde activity in the hippocampal tissue. Pretreatment with rapamycin, the mTOR inhibitor, abolished all the above mentioned effects of CM. These findings suggested that CM acted through the mTOR signaling pathways to regulate iNOS expression and oxidative state in the hippocampal tissue after I/R injury.

The plants of the genus Polygala (Polygalaceae) are employed in folk medicine for the treatment of several pathologies, including disorders of the bowel and kidney, as anesthetic, expectorant and anti-inflammatory. The present study was undertaken to investigate the antiedematogenic and antinociceptive activities of methanolic extract of Polygala boliviensis A. W. Benn (MEPB) in mice. The antinociceptive activity of MEPB was evaluated using the writhing, formalin and tail immersion tests. The carrageenan-induced paw edema test was used to assess the antiedematogenic activity of MEPB. Mice motor performance was evaluated in the rota rod and open field tests and the acute toxicity evaluated over 14 days. High-performance liquid chromatography was used to determine the fingerprint chromatogram of MEPB. Oral administration of MEPB (75- 600mg/kg) reduced the number of writhing induced by acetic acid. In the formalin test, the oral pre-treatment with MEPB (75 - 600 mg/kg) produced a dose-related inhibition only of the late phase. MEPB (300 and 600 mg/kg) reduced the carrageenan-induced paw edema. In contrast, the treatment with MEPB (300 and 600 mg/kg) did not prevent the thermal nociception in the tail immersion test. MEPB (600 mg/kg)-treated mice did not show any motor performance alterations. Over the study duration of 14 days, there were no mortality or toxic signs recorded in the group mice given 6000 mg/kg of MEPB. The present study demonstrates, for the first time, the antinociceptive and antiedematogenic properties of Polygala boliviensis.

The objective of this study is to investigate the anti-tumor effect of Ginkgo biloba exocarp extracts (GBEE) on B16 melanoma bearing mice and its related molecular mechanisms. The B16-F10 melanoma solid tumor model was established in C57BL/6J mice. The tumor-bearing mice were treated with GBEE (50, 100, 200 mg/kg), taking cis-Dichlorodiamineplatinum (Ⅱ) (DDP, 3 mg/kg) as positive control and normal saline (NS) as model control. After 17 days of administration, the transplanted tumors was stripped and weighed, and the inhibition rate was calculated. Quantitative Reverse transcription Polymerase chain reaction (qRT-PCR), Western Blot and immunohistochemistry were applied to detect mRNA and protein levels of related factors in B16 transplanted tumor tissues. The results indicated that GBEE (50, 100, 200 mg/kg) inhibited the growth of B16 transplanted solid tumor in C57BL/6J mice. Meanwhile, it inhibited the expression of CD34 and reduced microvessel density (MVD) in a dose-dependent manner. Moreover, GBEE dose-dependently down-regulated the mRNA and protein levels of hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), and vascular endothelial growth factor receptor 2 (VEGFR2). The phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt) proteins were not changed obviously, but the protein levels of p-PI3K and p-Akt were down-regulated. Overall, the inhibitory effect of GBEE on the growth of B16 melanoma transplant tumor in mice is related to inhibiting angiogenesis, and the mechanism involves the regulation of PI3K/Akt/ HIF-lα/VEGF signaling pathway.

Inflammatory reactions are closely associated with the development and progression of epilepsy. It has been shown that inhibition of pro-inflammatory cytokines, which are released from activated astrocytes and microglia, are considered to be an effective therapeutic approach for the treatment of epileptic disorders. Regarding the anti-inflammatory effects of nutmeg (Myristica fragrans Houtt), the present study was designed to investigate whether the nutmeg ethanolic extract could exert anticonvulsant and inhibitory effects on glial activation in pentylenetetrazol (PTZ) induced mice model of kindling. Ethanolic extract of nutmeg was administrated intraperitoneally (i.p) 1 hour before PTZ injection or one week before PTZ as a separate group, to become fully- kindled. The chemical components of nutmeg extract were analyzed by gas chromatography mass spectrometry (GC-MS). Immunostaining against neuronal and glial markers was performed on hippocampus sections. GC-MS data indicated that the main components of nutmeg extract are myristic acid (39.93%), elemicin (22.16%) and myristicin (11.17%). Behavioral studies showed that pre-treatment of nutmeg extract effectively reduced seizures behavior, decreased cell death, and ameliorated glial activation that is followed by PTZ administration. In conclusion, nutmeg extract might be regarded as a useful supplementary agent in epilepsy treatment through its attenuation of neuronal loss and glial activation.

Nitric oxide (NO) is a key mediator that plays an important role in pathogenesis of various chronic diseases like Alzheimer’s disease and Parkinson’s disease. Additionally, there is a great attitude for finding natural compounds, which could control and inhibit NO production in pathological conditions. Therefore, we were encouraged to investigate the effects of some Lamiaceae species on NO production and cell injury during oxidative stress in PC12 cells. In this study, cell death determined by MTT assay and NO levels were evaluated using Griess assay. PC12 cells were exposed to total metanolic extracts of three Scutellaria and one Nepeta species. The results revealed that Nepeta laxiflora (N. laxiflora) could protect PC12 cells from hydrogen proxide-induced oxidative stress and all of the plants inhibited NO production in that condition except Scutellaria tournefortii (Sc. tournefortii). In addition, Scutellaria multicaulis (Sc. multicaulis)was meanwhile subjected to fractionation using different organic solvents. The dichloromethan and ethyl acetate fractions of Sc. multicaulis could protect PC12 cells from oxidative stress injury. However, NO production was restrained by the hexane and dichloromethane fractions. Considering the results, N. laxiflora, Scutellaria nepetifolia (Sc. nepetifolia), and Sc. multicaulis are good candidates for further investigations in neuroprotection and anti-inflammation studies.

Noscapine is a natural alkaloid with anti-angiogenesis activities. The aim of the present study was to examine the effect of noscapine on eutopic endometrium of endometriosis patients (EEE) and normal endometrium (NE)in a three-dimensional (3D) culture model. In this experimental in-vitro study, EEE (n = 8) and NE (n = 8) biopsies were taken from 16 reproductive aged women. The biopsies were cleared from blood and mucus. Each biopsy was cut into small fragments (1 Í 1 mm) in a sterile condition. For 3D culture, the endometrial fragments were put between two layers of fibrin jell made of fibrinogen solution [3 mg/mL in Medium199 (M199) + thrombin]. Twenty-four wells of culture dish was divided into 5 groups for each biopsy: the control wells were treated with M199 containing 5% fetal bovine serum (FBS) while, the test wells were exposed to the same media containing one of the noscapine doses (10, 50, 100, and 200 μM). The expression of apoptotic genes, growth score, angiogenesis, and nitric oxide (NO) secretion were evaluated. The mean of growth score of groups exposed to 0, 10, 50, 100, and 200 μM were 2.2 ± 0.55, 1.7 ± 0.45, 1.44 ± 0.27, 0.29 ± 0.1, and 0.1 ± 0.08 in EEE, and also, 2.11 ± 0.6, 1.65 ± 0.5, 0.79 ± 0.41, 0.18 ± 0.1, and 0.1 ± 0.1 in NE, respectively, and the difference between the groups was significant (P < 0.05). The expression of apoptotic genes significantly increased while, the levels of Bcl-2 and Sirt1 reduced (P = 0.004). NO secretion reduced significantly (P < 0.05) in both EEE and NE groups. In conclusion, higher doses of noscapine showed inhibitory effect on growth and angiogenesis of EEE and NE.

The present study was designed to investigate the in-vitro morphological assessment of apoptotic effect caused by nimbolide on the selected cancer cell lines (DU-145, PC-3, A-549) and normal cell lines (NIH3T3, CCD-18Co). The cells were grown in 6 well tissue culture plates after treatment with different concentrations of nimbolide and untreated control cells. Apoptotic and necrotic cells were measured using Hoechst 33342 and propidium iodide dual staining through a fluorescent microscope and also by staining with annexin V and propidium iodide through flow cytometric analysis. The activity of caspase 3, 8, and 9 was measured by caspases colorimetric assay kits. The number of apoptotic and necrotic cells were significantly higher in all selected cancer cell lines treated with nimbolide as compared with untreated control cells, whereas in normal cell lines no significant difference was observed between nimbolide treated and untreated cells. The activity of caspase 3, 8, and 9 was also significantly higher in all cancer cell lines treated with nimbolide as compared with untreated control cells while it did not change significantly in normal cell lines as compared with untreated control. The results of the present study suggested that nimbolide induced apoptosis only in cancer cells without affecting the normal cells and one of the apoptosis inducing mechanism is through the activation of caspases signaling pathways. Therefore, nimbolide may be a novel promising candidate as an anticancer drug in future.

Tuberculosis (TB) is leading cause of death worldwide from infectious diseases and its inadequate treatment has led to emergence of resistant strains. The emergence of these strains renders the search for new drugs for the treatment of TB. The aim of this study was the evaluation of the anti-TB activity of the extract from fungus Gliocladium sp. MR41, and bioassay-guided fractionation and identification of majority compounds was carried out. Fungal strain culture was lyophilized and extracted by maceration in Ethyl Acetate (EtOAc). This extract was fractionated by liquid-liquid partitioning and chromatographic techniques, and the compounds were identified by their spectroscopic data. Furthermore, the EtOAc extract, fractions, and pure compounds were tested on Mycobacterium tuberculosis using the Microplate Alamar Blue Assay. From the bioactive AcetoNitrile Fraction (AcNF; MIC = 3.13 µg/mL) of the EtOAc extract, four compounds were isolated: ergosterol (1), ergosterol-5,8-peroxide (2), 1,6-di-O-acetyl-2,3,4,5-tetrahydroxy-hexane (3), and allitol (4). Only 2 exhibited potent activity against M. tuberculosis (MIC = 0.78 µg/mL). Additionally, this is the first report, to our knowledge, of polyols 3 and 4 from this fungus.

The decoction of Pheretima aspergillum is used as a traditional medicine for treatment of asthma in China with a long clinical history. The aim of the study was to investigate the anti-asthma activities of an active components group obtained from the decoction of Pheretima aspergillum in histamine and ovalubumin induced asthma guinea pigs. The incubation period of asthma, serum IFN-γ, IL-4 and LTB4 levels were tested and determined. As a result, the significant increase of asthma incubation period, serum IFN-γ level were observed in the asthma guinea pigs treated with the active components group. In addition, the serum IL-4 and LTB4 levels were obviously decreased compared to that of model controls. The chemical constituents of the active components group were identified or characterized using liquid chromatography-quadrupole time of flight mass spectrometry (LC-QTOF-MS/MS). Twenty oligopeptides were first identified or characterized in the active components group from the decoction of Pheretima aspergillum. It is reasonable to assume that the anti-asthma activities of Pheretima aspergillum mainly resulted from these oligopeptides. Also, synergistic effects among their compounds may exist in the anti-asthma activity of Pheretima aspergillum.

Physiological studies confirm improvement of memory by Olibanum, a resin from Boswellia species, while little is known about the molecular mechanism by which it affects memory performance. Two master transcription factors, CREB-1 and CREB-2, regulate downstream memory-related genes expression, leading to the long-term memory potentiation. This study addresses the effects of Beta-boswellic acid (β-BA), the main ingredient of Olibanum, and ethanolic extract of the resin from Boswellia serrata on the expression of CREB-1 and CREB-2 genes in B65 cell line. B65 cells were treated with β-BA) or ethanolic extract of Olibanum in different dose and time intervals and the cell viability/toxicity was measured by MTT assay. Total RNA was extracted from the treated and untreated control cells and cDNA was synthesized. The expression levels of CREB-1 and CREB-2 genes were quantified by Real-time PCR. MTT assays revealed 50% inhibitory concentrations of 42.05, 29.63 and 21.78 μg/ml for ethanolic extract of Olibanum and 89.54, 44.05 and 21.12 µM for β-BA at 24, 48 and 72h time intervals respectively. Both β-BA and ethanolic extract of Olibanum altered CREB-1 and CREB-2 genes expression levels in time-dependent but not in dose-dependent way. However, β-BA showed stronger and more stable effects. The expression levels of the both genes followed an alternate upregulation and downregulation pattern, but in opposite directions, in response to the both solutions with the progress of time. These results suggest that Olibanum possibly improves memory performance, at least partially, by regulating the levels of CREB-1 and CREB-2 transcription factors via positive/negative-feedback loops.

In this study we examined enhancement effects of Artemisinin plus Glucantime and shark cartilage extract, on promastigotes and amastigotes of L.infantum in in vitro condition.The toxicity of artemisinin, glucantime and shark cartilage extract on the L. infantum promastigotes and amastigote-infected macrophages was evaluated using MTT assay. The role of these drugs in inducing apoptosis in promastigotes and parasite-infected macrophages was also studied. Using promastigote assay, IC50 values of artemisinin and glucantime as stand alone drugs as well as in combination were obtained to be 50, 400 and 100µg/ml respectively. The flow cytometry analysis of apoptotic promastigotes stained with Annexin-V FITC staining showed that artemisinin, glucantime, artemisinin plus glucantime, artemisinin plus shark cartilage extract and shark cartilage extract applied at their IC50 concentrations resulted in 53.5%, 73.92%, 64.46%, 49.9% and 47.34% apoptosis respectivelyThe results of MTT assay indicated that cytotoxicity of artemisinin, glucantime, artemisinin plus glucantime, shark cartilage plus artemisinin and shark cartilage in infected macrophages after 72hrs was 75%,84%,82%,30% and 3% respectively. In non- infected macrophages, cytotoxicity of Artemisinin, Glucantime, Artemisinin plus Glucantime and shark cartilage was 15%, 31%, 21%,2% and 0% respectively.This study suggests that artemisinin, glucantime, artemisinin plus glucantime and shark cartilage extract have significant killing effects on promastigotes and amastigotes. Also, it proved that artimisinin alone and in combination with glucantime and shark cartilage extract has little toxic effect on macrophages, but could induce apoptosis in L.infantum promastigotes and amastigote-infected macrophages Thus these chemicals can be used as alternative drugs for in vivo studies.

Sideritis species have been known as medicinal plants since ancient times, and used as tea in Mediterranean countries such as Turkey, Greece and Spain. They are also used for the treatment of several ailments such as a cough, common cold and gastrointestinal disorders. The aim of the present study was to perform the chemical composition, antioxidant, anticholinesterase and anti-tyrosinase activities of the essential oils of Sideritis albiflora and S. leptoclada. β-caryophyllene (21.2%) and Germacrene D (17.9%) were identified as the major compounds in S. albiflora and S. leptoclada essential oils, respectively. The essential oil of S. albiflora showed the highest lipid peroxidation inhibitory (IC50:73.8±0.8 µg/mL), DPPH free radical scavenging (28.3±0.1%), ABTS cation radical scavenging (IC50:50.6±1.0 µg/mL), reducing power (A0.05: 181.7±0.6 µg/mL), acetylcholinesterase (22.1±0.4%), butyrylcholinesterase (IC50:157.2±0.9 µg/mL) and tyrosinase (15.2±0.4%) inhibitory activities. Moreover, S. albiflora essential oil had rich total phenolic and flavonoid contents indicating 41.5±0.8 µg PEs/mg extract and 21.4±1.0 µg QEs/mg extract, respectively. This study suggests that consumption of Sideritis species as tea may protect one against melanogenesis, amnesia, and oxidative stress without any observable side effect.

A rapid and simple method has been developed for the screening and identification of natural antioxidants from the leaves of Acer ginnala Maxim.( AG). The process is that upon reaction with 1,1-diphenyl-2-picrylhydrazyl (DPPH), the white yellow spots of compounds with potential antioxidant effects will be significantly observed on the thin-layer chromatography (TLC), and possible structures will be presumed by the ESI/MS technique. Using the improved approach, 6 compounds in the AG extract were found to possess a potential antioxidant activity. They were speculated as quercetin-3-O-α-L-(3''-galloyl)-rhamnoside (1), quercetin-3-O-α-L-(2''-galloyl)-rhamnoside (2), quercetin-3-O-α-L-(2''-galloyl)-arabinopyranoside (3), acertannin (4), gallic acid (5) and methyl gallate (6). In addition, we were still found that compounds 2, 3 and 5 had favorable antioxidant activity from the scannogram of the DPPH reaction plate. As a result, the isolated 6 compounds structures were in accord with the presumed structures. Furthermore, the free radical scavenging capacities of the available identified compounds were also investigated. Compounds 2, 3 and 5 showed significant DPPH. scavenging capacities, with IC50 values of 2.83 μg/mL, 2.34 μg/mL, 1.86μg/mL, respectively. The results indicated that this newly improved method could be widely applied for rapid screening and identification of natural antioxidants from Chinese herbal medicines.

Despite the traditional use of Solidago canadensis L. (Asteraceae) as a diuretic drug, there is a scarcity in scientific data concerning the activity of its different extracts and fractions as well as the class of constituents responsible for this diuretic action. A comparative study was carried out for the diuretic activities of the different standardized extracts and fractions of the flowering aerial parts of S. canadensis, as well as isolation of compounds from the most biologically active fraction. The ethanol extract and its ethyl acetate fraction (EA) showed the highest aquaretic activities (91 and 58% at a dose of 400 mg/Kg b.wt., respectively) compared to 100% of furosemide at 20 mg/Kg b.wt.. Their activities were higher than Cystinol® and spironolactone reference standards (74% and 59% of furosemide, respectively). EA showed the highest total phenolic and flavonoid contents among the tested fractions of the ethanol and aqueous extracts (9.38 ± 0.004 g GAE and 39.75 ± 0.005 g RE/100 g dried extract, respectively). Eight flavonoids, 2 phenolic acids and 1 nucleoside were isolated from EA. This is the first report of a comparative study between the aquaretic activities of the different extracts, fractions and essential oil of S. canadensis, as well as isolation of thyimidine (1), isorhamnetin-3-O-β-ᴅ-glucopyranoside (2), kaempferol-3-O-(6"-O-acetyl)-β-ᴅ-glucopyranoside (4), quercetin-3-O-(6"-O-acetyl)-β-ᴅ-glucopyranoside (5), and kaempferol-3-O-β-ᴅ-apiofuranoside (7) from genus Solidago.

A new bioactive proline-rich cyclohexapeptide - diandrine C (6), previously isolated from whole plant of Drymaria diandra (Caryophyllaceae), was synthesized through coupling reactions of tetrapeptide unit Boc-Gly-l-Pro-l-Tyr-l-Trp-OH with dipeptide unit l-Pro-Gly-OMe using N,N-diisopropylcarbodiimide (DIPC) as the coupling agent, followed by cyclization of linear hexapeptide unit under alkaline condition. Structure of cyclohexapeptidewas confirmed by means of chemical, and spectroscopic analyses and also was screened for its antimicrobial and anthelmintic properties. Bioevaluation results indicated that the newly synthesized hexacyclopeptide exhibited potent antimicrobial activity against Gram-negative bacteria Pseudomonas aeruginosa, Klebsiella pneumoniae and pathogenic Candida albicans at 6 mg/mL. Moderate to good level of antihelmintic activity against three earthworm species Megascoplex konkanensis, Pontoscotex corethruses and Eudrilus eugeniea was also observed at concentration of 2 mg/mL.

FeiYangchangweiyan capsule (FY capsule), a traditional Chinese medicinal preparation consisting of three medicinal herbs, has been used to treat bacterial dysentery, acute, and chronic gastroenteritis for several decades. In this study, a novel, convenient, accurate, and valid method was developed by using high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD) to obtain a chromatographic fingerprint of FeiYangchangweiyan capsule (FY capsule). Then, fourteen peaks were identified according to MS/MS fragmentation behavior of the reference standards by using HPLC-DAD-ESI-MS/MS analysis. At the same time, the fingerprint similarity was calculated and the contents of known ingredients were also determined simultaneously. The result demonstrated that the HPLC fingerprint combining similarity evaluation and quantification analysis can be successfully applied to control the quality of FY capsule.

Streptokinase (SK) is an extracellular protein comprising 414 amino acids with considerable clinical importance as a commonly used thrombolytic agent. Due to its wide spread application and clinical importance designing more efficient SK production platforms worth investigatinginvestigation. In this regard, a synthetic SK gene was optimized and cloned in to pET21b plasmid for periplasmic expression. Response surface methodology was used to design a total of 20 experiments for optimization of IPTG concentration, post-induction period, and cell density of induction (OD600). The optimum levels of the selected parameters were successfully determined to be 0.28 mM for IPTG concentration, 9.889 H for post induction period, and 3.40768 for cell density (OD600). These settings result in 4.14 fold increase in SK production rate of optimum expression conditions (7663 IU/mL) in comparison to the primary expression conditions (1853 IU/mL). Achieving higher yields of SK production in shake flask could lead to more cost effective industrial production of this drug which is the ultimate aim of SK production studies.

Vitamin D deficiency causes osteoporosis, osteopenia, fractures, rickets and more recently is linked with some chronic illnesses such as cancer. Because of the safety and probiotic properties of the yeast Saccharomyces cerevisiae, we hypothesized that yeast cells enriched with cholecalciferol (vitamin D3) could represent a solution for prevention or treatment of vitamin D deficiency. In this study S. cerevisiae was used as a vitamin D3 accumulator for the first time and the optimal conditions for enrichment of S. cerevisiae was determined. The Plackett-Burman screening studies were used for selection of the most important factors affecting cholecalciferol entrapment. Response surface methodology was employed for optimization of cholecalciferol accumulation in S. cerevisiae cells by using Box-Behnken design. A modified quadratic polynomial model fit the data appropriately. The optimal points of variables to maximize the response were cholecalciferol initial concentration of 358021.16 IU/ml, tryptone concentration of 1.82 g/l, sucrose concentration of 7.13 % (w/v) and shaking speed of 140.46 rpm. The maximum amount of cholecalciferol in dry cell weight of S. cerevisiae was 4428.11 IU/g. The cholecalciferol entrapment in yeast biomass increased about two-folds in optimized condition which indicates efficiency of optimization.

Our aim was to examine the effect and cost-effectiveness of isosorbide mononitrate (IMN) for outpatient (at home) cervical ripening in Iranian women with term pregnancy. Term pregnant women were randomly selected to receive either  15 mg vaginally administered IMN or placebo. Patients were advised to take the second dose at home 24 h later and return to the hospital for admission after the next 24 h if the onset of spontaneous contraction does not occur. Cervical status, maternal blood pressure, maternal pulse rate, fetal heart rate and various side effects were examined. IMN induced a significant increase in cervical Bishop Score; headache, nausea, and dizziness were seen in 8 of 28 participants of IMN group; although, all had normal vital signs and normal FHR, they had to be hospitalized and receive medication to relieve the unpleasant symptoms. Vaginal administration of 15 mg of IMN induced cervical ripening in term pregnant women.  However, the administration of IMN for cervical ripening is not recommended as it produced headache in significant number of Iranian pregnant women.

Today, an increase in vancomycin dose has been proposed to ensure efficacy. However, the risk of nephrotoxicity will increase with the dose. This study was aimed at evaluating the dosage regimens of vancomycin in pediatric patients based on pharmacokinetics/pharmacodynamics (PK/PD) and to optimize dosage individualization. Population pharmacokinetics analysis was performed on 155 Chinese children (aged 1 month to 16 years), which were divided into various renal function subpopulations. Monte Carlo simulation was carried to evaluate the efficacy and safety of vancomycin dosage regimens on each subpopulation. Compared with children with normal renal function as glomerular filtration rate (GFR)≥90 mL/min•1.73m2, the clearance of vancomycin decreased by 39.4% and the half life increased 1.74 fold respectively in children with moderate renal inadequacy (30≤GFR＜60 mL/min•1.73m2). When vancomycin was administered as conventional dosage (40-60mg•kg-1•d-1) to against methicillin-resistant staphylococcus aureus (MRSA) with higher MICs of 1-2 mg•L-1 for children with normal renal function, the probability of efficacy target attainment( PTA) at AUC0-24h/MIC≥400 (where AUC is the area under curve and MIC is the minimum inhibitory concentration) achieved≤63.64%. While vancomycin dosage exceeded 70mg•kg-1•d-1 for children with normal renal function, 50mg•kg-1•d-1 for mild renal inadequacy (60≤GFR＜90 mL/min•1.73m2), 30mg•kg-1•d-1 for moderate renal inadequacy respectively, the PTA at trough concentration above 20 mg•L-1 achieved ＞20%, that not to be suggested for high risk of nephrotoxicity. Considering both efficacy and safety, the conventional vancomycin dosage is not enough and adjustable interval is narrow for againsting MRSA with MICs of 1-2 mg•L-1.

A high prevalence of genetic polymorphisms increases sensitivity to warfarin therapy. In this study, we investigated 47 patients with effective long-term therapy by warfarin well-controlled by monitoring of International Normalised Ratio (INR). All patients were tested for gene polymorphisms VKORC1, CYP2C9*C2, and CYP2C9*C3, which were used for a dose calculation employing a program www.WarfarinDosing.org. The main goal was to investigate whether the warfarin doses determined by INR are in accordance with the doses calculated according to the pharmacogenetic algorithm. For this purpose, several chemometric tools, namely principal component analysis, cluster analysis, correlation analysis, correspondence analysis, Passing-Bablock regression, Bland-Altman method, descriptive statistics, and ANOVA were used. We also analysed the relationship between the dose of warfarin determined by INR and several constitutional and genetic factors. Statistically significant association between clinically optimized warfarin dose and indication for the treatment, age, and warfarin sensitivity determined by VKORC1, CYP2C9 gene polymorphisms were confirmed. Finally, we confirmed a good concordance between the INR determined warfarin doses and pharmacogenetic approach.

The aim of this systematic review was to evaluate the effect of purgative manna on the unconjugated hyperbilirubinemia in neonates. Pubmed, Scopus, Chochrane library, Iranmedex and Google scholar were last searched in February 2017. Randomized controlled trials that evaluated the effect of purgative manna on the treatment of neonatal hyperbilirubinemia were included in the study. For meta-analysis, weighted mean difference (WMD) with 95% confidence interval (CI) was used. The outcomes of interests were serum bilirubin levels and length of hospital stay in neonates with jaundice. Seven randomized controlled trials with 812 neonates were eligible to be included in this systematic review. The meta-analysis included six of seven controlled trials. Bilirubin levels were significantly lower at 12 h (WMD : -1.48, 95% CI: -2.31 to -0.65), 24 h (WMD: -2.47, 95% CI: -3.22 to -1.71), 36 h (WMD: -2.83, 95% CI: -4.87 to -0.80), 48 h (WMD: -1.49, 95% CI: -2.36 to -0.63) and 72 h (WMD: -0.68, 95% CI: -1.28 to -0.08) following intervention in purgative manna group. Length of hospital stay was also decreased in purgative manna group (WMD: -0.93, 95% CI: -1.35 to -0.50). Finally, purgative manna administration decreased serum bilirubin level and length of hospital stay in neonates with unconjugated hyperbilirubinemia. More studies are needed to evaluate the efficacy, dosage, and side effects of purgative manna.

Gastric cancer is the fourth common cancer and the second leading cause of cancer death worldwide. Due to lack of adequate information on the side effects of chemotherapy regimens in treatment of gastric cancer, this study was aimed to determine the side effects of two common chemotherapy regimens of gastric cancer. This prospective study was conducted in Emam Khomeini educational hospital and Touba Polyclinic, both are affiliated to Mazandaran University of Medical Sciences. The frequency and severity of side effects of chemotherapy were recorded based on the National Cancer Institution (NCI) Toxicity Criteria (version 2). DCF (Docetaxel, Cisplatin, 5FU) and FOLFOX (Folinic acid, 5FU, Oxaliplatin) adverse reactions were compared using SPSS 16 software. One hundred twenty five chemotherapy cycles administered to seventy four patients were assessed. The most common used regimens were DCF (70%) and FOLFOX (16%). The incidence of vomiting was higher with DCF compared to FOLFOX (P=0.049). In more than 50% of cycles, DCF regimen caused diarrhea, while in FOLFOX regimen it was less than 9% (P=0.002). Stomatitis, visual changes, nausea, skin reactions and constipation were not significantly different between the two regimens. It seems that the adverse drug reactions of FOLFOX regimen were more favorable than DCF regimen. The results of this study may help clinicians choosing a more favorable chemotherapy regimen especially in patients with a low performance status who have difficulties in tolerating a chemotherapy regimen with a more severe adverse effect profile.

Contrast-induced nephropathy (CIN) (known as contrast-induced acute kidney injury) occurs as a result of acute worsening of renal function following a procedure with administration of iodine contrasts agent and remains a substantial concern in clinical practices. The purpose of this study is to investigate the preventive effect of Pentoxifylline supplementation on reduction of CIN occurrence after percutaneous coronary intervention among patients who were high risk of CIN according to Mehran score.In randomized, double-blind clinical trial patients who undergo coronary angiography with Mehran Score ≥ 11 consisted of our population. Patients in a ratio 1:1, divided into two groups received saline 0.9% plus N-acetyl cysteine and Pentoxifylline 400 mg three times per day 24 h before angiography until 48 h after angiography. In control group, the patients received placebo instead of PTX in a same manner as the control group. The endpoint was the incidence of CIN defined as creatinine increase of 0.5 mg/dL within 2 days after contrast.There were no significant differences in baseline characteristics. CIN occurred in 3 (5.5%) and 4 (7.3%) patients of the both groups (Pentoxifylline and control), respectively (p = 0.69; incidence odds ratio 1.36; 95% CI 0.29-6.38). No significant differences were seen in secondary outcome measures and changes in the level of creatinine (p = 0.54).In high-risk patients undergoing coronary angiography pentoxifylline supplementation had protection effect against contrast-induced nephropathy greater than placebo based hydration, but, not supported by our data.

Usage of cancer chemotherapeutics drugs can be associated with adverse drug reactions. When IgE-mediated drug reactions are formed following administration of a chemotherapeutic drug that is a drug of choice, drug desensitization protocols can be helpful. Hypersensitivity reactions can be allergic or nonallergic, but the clinical manifestations are similar. Rapid drug desensitization (RDD) is effective when used appropriately, however it is often over utilized instead of performing a drug challenge. RDD is both an acceptable approach and a high-risk treatment modality in patients in whom the first line chemotherapy is the offending agent. The sa

The aim of this study was to assess the correlation of serum vitamin D with ICU length of stay, mortality rate, length of mechanical ventilation, and incidence of sepsis. We conducted a descriptive analytic study on 793 patients admitted to surgical ICU wards in northwest of Iran from March 2015 to March 2016. Patients were assessed during the ICU stay and the following data were collected: Glasgow Coma Score (GCS), APACHE II score, incidence of sepsis, duration of mechanical ventilation, LoS, mortality rate, and laboratory data (such as serum vitamin D, calcium, phosphorus, etc). The effect of vitamin D deficiency and the confounding factors on length of stay was assessed using the multinomial regression. Of 793 patients, 161 patient (20.3%) were in vitamin D deficiency group, 306 (38.6%) in vitamin D insufficiency group, 326 (41.1%) in vitamin D sufficiency group. Vitamin D deficiency increased risk of sepsis( OR=22.93; 95%CI :10.631-49.78 ) and mortality rate (OR=42.93; 95%CI: 15.2-121.22) . Vitamin D deficiency/insufficiency is a result of chronic and severe comorbidities of patients and can be considered as a helper but not a real risk factor for mortality and its level should be assessed in surgical critically ill patients. However, various levels of vitamin D impact outcome in critically ill patients remains to be elucidated and further multi-center trials are needed to validate our result

Autophagy, known as cell death type II, is a housekeeping pathway that currently has been worked on in matters of tumorigenesis and leukemogenesis. Therefore, in this study expression levels of ATG7 and LC3 as two key genes are targeted in AML patients. This study was performed on 55 de novo AML patients against 17 healthy volunteers, acquired samples from bone marrow (BM) and peripheral blood (PB) sources in different ages and gender. The evaluation was executed by mRNA extraction, cDNA synthesis, real-time PCR and data was analyzed by SPSS. Analyzed data indicate a significant decrease between expression of ATG7 and LC3 in AML patients against control (Pv< 0.05). Decrease in both genes expression was detected in most of the patients, 81.81% and 75.55%, respectively. Also LC3 overexpression was detected in 11.33% of AML patients. Moreover, a positive significant correlation between ATG7 and LC3 genes was detected (r= 0.481; Pv= 0.001). This study showed that significant reduction of autophagy genes in de novo AML patients is important to overcome this system and initiate leukomogenesis. It seems a new insight is required for new achievements in diagnosis, prognosis, treatment and monitoring AML patients.

The relationship of vitamin D3 with the duration of mechanical ventilation and mortality is still unknown. Therefore, this study aimed to determine the effect of using high-dose vitamin D on the duration of mechanical ventilation among the patients admitted to the intensive care unit. The current double-blinded clinical trial was performed on 44 mechanically ventilated, adult patients. Using permuted block randomization, the patients were recruited in intervention and placebo arms. In the placebo group, four patients were excluded due to death before 72 h. The vitamin D level was measured in both groups on entrance and 7th day of the study. The intervention and placebo groups received intramuscular injection of 300000 IU vitamin D and identical placebo, respectively. SOFA and CPIS score were evaluated daily for 7 days and on 14th and 28th days of the study. Also duration of mechanical ventilation and mortality rate were recorded. Fourteen males and 8 females were recruited in the intervention group, as well as 13 males and 5 females in the control group. There was no significant difference in baseline characteristics of the patients including gender and age. The mean duration of the mechanical ventilation was 17.63 ± 14 days in the intervention group versus 27.72 ± 22.48 days in the control group (p = 0.06). Mortality rate in control and intervention groups was 61.1% versus 36.3% (p = 0.00), respectively. Administration of high-dose vitamin D could reduce mortality in mechanically ventilated patients. Despite decrease of 10 days in duration of mechanical ventilation, the difference was not statistically significant. Larger studies are recommended.

Teriparatide is a new agent serves as a treatment of choice for severe post-menopausal osteoporotic patients who are at a high risk of fracture or have failed or been intolerant of previous osteoporosis therapy. The objective of this study is estimating the cost-utility of Teriparatide compared with no treatment from health system perspective in Iran. A microsimulation model was developed for a cohort of hypothetical Iranian patient population (women aged 70 years, T-score -2.5 with previous fracture or T-score -3.0 without prior fracture) over lifetime horizon. The model consisted of seven health states. During each cycle, patients could have a fracture, remain healthy, remain in a post-fracture state or die. Background fracture risks, mortality rates, persistence rates, utilities, medical and drug costs were derived using published sources. Total accumulated life-time costs and quality-adjusted life years (QALYs) were estimated. Teriparatide was associated with 4.786 QALYs and total direct costs of IRR 143,168,259 over a lifetime horizon. Compared to no treatment, Teriparatide provided an additional 0.145 QALY at an incremental cost of IRR 33,511,013. The resulting incremental cost-effectiveness ratio was IRR 230,333,030/QALYs gained. The probabilistic analysis showed that accepting a willingness-to-pay 2 and 3 GDP/capita in Iran, the probability of Teriparatide being cost-effective were 51% and 83%, respectively. Compared to no treatment, Teriparatide was indicated to be more costly and associated with fewer fractures, more life-years, and more QALYs. The result showed that Teriparatide may be considered a cost-effective intervention when targeted to the appropriate patients.

Traumatic brain injury (TBI) is a public health problem worldwide. Secondary damage of brain injury begins within a few minutes after the trauma and can last a long time. It can be reversible, unlike primary injury. Therefore, therapeutic intervention can be used. The aims of this study were to assess the effects of minocycline on neurological function and serum S100B protein and neuron-specific enolase (NSE) levels in patients with moderate to severe TBI. Patients with acute onset of TBI and surgical evacuation of hematoma were randomized to receive either minocycline 100 mg orally twice daily or placebo for 7 days. The primary outcomes included changes in level of S100B and NSE at different time points during the trial. Additionally, changes in Glasgow coma scale (GCS) score were evaluated. The Glasgow Outcome Scale-Extended (GOS-E) score at 6 month after injury was assessed in discharge patients. Thirty four patients were randomized into the placebo (n=20) and treatment (n=14) groups. There was a marginal statistically significant differences in the normalized value of S100B between groups (p<0.1). The reduction in serum NSE level from baseline to day 5 was statistically significant (p=0.01) in minocycline group while it was not significantly decrease in placebo group (p=0.2). Also, GCS improvement over time within the minocycline group was significant (p=0.04) while was not significant in placebo group (p=0.11). The GOS-E scores were not significantly different between minocycline and placebo group. Based on this study, it seems that the use of minocycline may be effective in acute TBI.

CYP2C19 polymorphism is associated with pretreatment drug response prediction, metabolism, and disposition. Pakistan consists of a population comprising of various ethnic groups residing in different regions of the country each claiming diverse ethnic origins. The identification of CYP450 genotypic composition of these populations is therefore necessary to avoid adverse drug reactions in these individuals. The main objective of the study was to investigate the prevalence of CYP2C19*2 and CYP2C19*17 alleles in these ethnic groups. The study was conducted on one thousand and twenty-eight (n = 1028) healthy volunteers from nine ethnic groups of Pakistan namely Brusho (n = 28), Hazara (n = 102), Kalash (n = 64), Pathan (n = 170), Punjabi (n = 218), Saraiki (n = 59), Brahui (n = 118), Parsi (n = 90), and Sindhi (n = 179). DNA was extracted from leukocytes and analyzed by allele specific amplification polymerase chain reaction (ASA-PCR). Multi allelic polymorphism of CYP2C19 led to four distinct phenotypes identified as extensive metabolizer (EM), poor metabolizer (PM), intermediate metabolizer (IM), and ultra-rapid metabolizer (UM). Over all, the percentage of predicted poor metabolizer allele was 29.0% compared to UM allele (23.70%).Among the studied groups, Saraiki and Brahui showed highest percentage of PM allele (40%, 36%) whereas Parsi and Hazara had highest percentage of UM allele (37% and 30% respectively). In conclusion, the high allele frequency of PM (CYP2C19*2 and *17) in Pakistani population led to the recommendation of a pre-treatment test to monitor drug response and dosage (personalized medicine) to avoid post-treatment adverse drug reactions.

In today’s competitive market environment, pharmaceutical companies have learned that improving supply chain performance is critical to maintain competitive advantages. Forecasting, planning, procurement, financing, stock levels, and marketing strategies are some of the areas in which managers have to decide about them and balance their enter-related effects simultaneously, to achieve organizational goals. This study is based on the results of literature review, experts’ opinion acquisition, and qualitative system dynamics modeling. So, according to method, triangular researches have been considered. The purpose of this research is to explore pharmaceutical supply chain (PSC) challenges and the dynamics behavior of variables playing a special role in PSC. Also, it provides different policies to overcome the challenges. For the first step to reach this goal, several semi-structured interviews with expert supply chain managers are conducted to explore the main challenges. Inaccuracy in forecasting, long lead times, lack of optimum target inventory, and high SC costs are the most important PSC problems. Then, qualitative system dynamics methodology is used to demonstrate the inter-relationship between variables that have impact on challenges. Finally, three strategic policies are recommended including: Collaborative relationship with suppliers, Investment in new technologies, and2Information technology (IT) establishment. Consequently, the results can give PSC managers a comprehensive view for decision making and bringing their attention to the importance of feedback behavior of variables in long term and their effects on organizational decisions and goals.

Dyslipidemia is responsible for great mortality and morbidity each year. Little data are available on the availability and affordability of Dyslipidemia medications in low and middle incomes countries. In a retrospective time-series study, we examined the utilization pattern and affordability of lipid-lowering medications in Iran as a lower middle-income country. We initially calculated the defined daily dose for 1000 inhabitants (DID) in different years and compared the results with OECD member countries in the same year. We also used 90% Drug Utilization method to rank and compare lipid lowering drugs with the WHO Essential Medicines List (EML). We measured the affordability by the minimum daily wage for one-month course of treatment. The use of lipid-lowering medications increased from 6.31 to 45.98 DID between 2005 and 2016. The utilization share of the subgroup of statins was above 80% of total utilization. Compared to OECD countries, Iran utilized 40% of the average utilization in 2015. In 2015, Atorvastatin was on 90% of DU medications. At the beginning of the study, only Lovastatin and Nicotinic acid were affordable in 2005, but at the end of the study, all lipid-lowering medications were affordable. The utilization of lipid-lowering medications, despite being affordable, was low. One of its possible reasons is the lack of proper management of patients with Dyslipidemia and low adherence of patients. Another possible cause is the high percentage of undiagnosed patients in the community. Therefore, comprehensive planning and policy-making should be taken to increase utilization and eliminate the related obstacles.

Almost all countries are affected by a variety of drug-supply problems and spend a considerable amount of time and resources to address shortages. The current study aims to reach a consensus on the scarce drug allocation measures to improve the allocation process of scarce drugs in Iran by a population needs-based approach. To achieve the objective, two phases were conducted. Firstly, a set of population-based indicators of needs were identified by reviewing the literature and were scrutinized by fifty academics/executives who were specialists in pharmaceutical resource allocation. In the second phase, the Delphi technique was performed to finalize the indicators. The yield of literature review step was about 20 indicators. Based on the results of the first questionnaire, 13 indicators were added to the Delphi phase. Then, in Delphi phase, the consensus was built after three Rounds. In addition to the burden of endemic, special, rare, and incurable diseases, traumatic diseases and total population of each province were the main measures. Furthermore, total mortality rates and the number of pharmacies in each province were on the border; hence, the monitoring team made the decision about inclusion or exclusion of such indicators. Other measures were in the range of 'important' ones. To reach a higher effective and efficient process of resource allocation, the paper suggests the use of a population needs-based approach in Iran's pharmaceutical sector. The scarce drug allocation indicators extracted in this study can make a considerable contribution to preventing, controlling, and mitigating drug shortages.