Tobramycin is one of the aminoglycoside antibiotics that lack a UV absorbing chromophore. However, the application of pulsed electrochemical detection (PED) has been used successfully for the analysis of this and simi...Tobramycin is one of the aminoglycoside antibiotics that lack a UV absorbing chromophore. However, the application of pulsed electrochemical detection (PED) has been used successfully for the analysis of this and similar antibiotics. This work describes an improved liquid chromatographic (LC) method combined with PED, which is able to separate much more impurities than before. Using a Discovery C-18 RP column (250 mm 4.6 mm i.d., 5 mm), isocratic elution was carried out with a mobile phase, containing sodium sulfate (35 g/L), sodium octanesulphonic acid (1 g/L), tetrahydrofuran (14 mL/L) and 0.2 M phosphate buffer pH 3.0 (50 mL/L). Using these experimental conditions, the limit of quantification (LOQ, S/N 10) was 5 ng. The linearity was examined in the range LOQ-60 mg/mL and the coefficient of determination was 0.998. The method also proved to be repeatable and the recovery was close to 100%. The influence of the different chromatographic parameters on the separation was investigated by means of an experimental design. The proposed method is useful in quality control of tobramycin drug substances and drug products.展开更多
Cefuroxime Axetil (CA) a widely used cephalosporin antibiotic displays low aqueous solubility and high membrane penetrability. This results in its solubility driven variable and/or low oral bioavailability and therape...Cefuroxime Axetil (CA) a widely used cephalosporin antibiotic displays low aqueous solubility and high membrane penetrability. This results in its solubility driven variable and/or low oral bioavailability and therapeutic efficacy as a major drawback. Thus, most of the goal of our study was to increase the solubility as well as dissolution rate of CA using the simple and cost-effective solid dispersion (SD) method. At first, the SD formulations of CA were prepared at various weight ratios of Carplex-67 and PEG-4000 by solvent evaporation technique. These new formulations were then subjected to an in-vitro drug release performance study and tested for physicochemical characterization to distinguish the thermal behavior, crystallinity, interactions phenomena, and surface morphology. Among the formulated Cefuroxime Axetil Solid Dispersion (CSD), CSD-8 which contained CA, Carplex-67, and PEG-4000 at the weight ratio 1:3:2, respectively showed the most significant (p in-vitro dissolution in water, Gastric Simulated Fluid (GSF), and Intestinal Simulated Fluid (ISF). This study also showed a significant (p < 0.001) increase in drug release compared to the marketed product. Therefore, it is supposed to be a promising alternative to conventional antimicrobial therapy.展开更多
Objective:Liver cirrhosis is a disease that seriously damages human health.Traditional Chinese Medicine(TCM)formulae have a good therapeutic effect on cirrhosis,and the herb pair is the smallest unit in formula compat...Objective:Liver cirrhosis is a disease that seriously damages human health.Traditional Chinese Medicine(TCM)formulae have a good therapeutic effect on cirrhosis,and the herb pair is the smallest unit in formula compatibility,which is important for improving the therapeutic effect.Therefore,identifying core herb pairs among TCM formulae is key.Methods:We mined the data of TCM formulae for the treatment of cirrhosis in the China National Intellectual Property Administration for the first time and analyzed their herb characteristics and association rules.We screened 405 patented TCM formulae,including 953 herbs.Based on frequency statistics and association rules,we determined“Astragali Radix-Salviae Miltiorrhizae Radix Et Rhizoma”as the core herb pair.Results:Six active compounds,Isorhamnetin,Formononetin,Calycosin,Cryptotanshinone,Dihydrotanshinone I,and Tanshinone II A,were screened out based on previous studies and network pharmacology.We found that SRC,TP53,HSP90AA1,MAPK3,MAPK1,and STAT3 played pivotal roles in treating cirrhosis.Interestingly,molecular docking indicated that MAPK3 might be a potential pharmacological target for cirrhosis.Conclusion:We preliminarily predicted and verified the pharmacological and molecular mechanism of“Astragali Radix-Salviae Miltiorrhizae Radix Et Rhizoma”in treating cirrhosis.This can expand the scope of TCM in the treatment of cirrhosis,guide people to use clinical formulae,and provide valuable insights for further drug discovery studies.展开更多
Paracetamol (PCM) is enlisted in the WHO model list as an essential medicine for pain and palliative care, but at overdose, it causes hepatic damage. This study was designed to assess the analgesic efficacy and hepato...Paracetamol (PCM) is enlisted in the WHO model list as an essential medicine for pain and palliative care, but at overdose, it causes hepatic damage. This study was designed to assess the analgesic efficacy and hepatoprotective property of a solid dispersion (SD) loaded with PCM. A number of PCM loaded formulations (PSDs) were fabricated using silica alone or in combination with polyethylene glycol and/or Na-citrate followed by in-vitro dissolution profiling. Selected PSDs with improved dissolution profile were subjected to solid-state characterization (DSC, PXRD, FTIR, and SEM), stability study along with investigation of in-vivo analgesic efficacy and effect on hepatocytes. Among these, PSD10 showed a rapid and significantly higher in-vitro drug release than pure PCM. This improvement was distinct to other PSDs also. Solid-state characterization of PSD10 authenticated the conversion of crystalline PCM to amorphous form upon formulation. Subsequent oral administration of PSD10 in Swiss albino mice showed 1.44-fold greater analgesic efficacy than pure PCM at dose 30 mg/kg. Besides, at acute toxic dose, liver histology of PSD10 mice was comparable with NC mice indicating hepatic protection upon formulation, whereas the PCM mice showed extensive hepatic necrosis which was also endorsed by significantly higher values of SGPT, SGOT, and ALP than PSD10 mice. Finally, an accelerated stability study of PSD10 performed according to the guideline of ICH noticed no remarkable deviation in its dissolution performance as well as crystalline nature. Thus, this newly developed PSD10 may be a safe and promising alternative for pain management and palliative care.展开更多
Objective:To observe the effect of co-culture cytokine-induced killer cells(CIK) and homologous dendritic cells(DC) on the proliferative activity and phenotype change of the DCCIK cell and the cell killing activity of...Objective:To observe the effect of co-culture cytokine-induced killer cells(CIK) and homologous dendritic cells(DC) on the proliferative activity and phenotype change of the DCCIK cell and the cell killing activity of leukemia HL-60.Methods:50 mL cord blood sample was obtained from infants delivered by full term healthy woman and the cord blood mononuclear cells were isolated by density gradient centrifugal ion.Non-adherent cells were collectedfor the induction culture of CIK.adherent cells were differentiated into mature DC;cultured mature DC was mixed with and CIK in the proportion of 1:5 for 12 d.killing activity of DC-CIK co-cultured cell on leukemia HL-60 was detected by MTT assay.Results:Compared with CIKs.the cocultured DC-CIKs presented a markedly higher proliferation and killing activity.Conclusions:Co-culture of DC-CIK cells led to a significant increase of the proliferation and cytotoxicity of CIK.展开更多
Background:Vine tea from fermented Ampelopsis grossedentata leaves has been used as a herbal tea and folk medicine in the southern region of China for hundreds of years.The aim of this investigation was to analyze the...Background:Vine tea from fermented Ampelopsis grossedentata leaves has been used as a herbal tea and folk medicine in the southern region of China for hundreds of years.The aim of this investigation was to analyze the total flavonoids found in vine tea,including three bioactive flavonoids,and the total phenolic contents in the aqueous methanol extracts of 10 vine tea samples.In addition,this study also aimed to examine the antioxidant activity of dihydromyricetin and vine tea’s flavonoid-rich extract.Methods:The total flavonoids and total phenolic content assay of extracts from vine tea were performed by ultraviolet-visible spectroscopy and epoch microplate spectrophotometer,respectively.Three bioactive flavonoids were quantified simultaneously using high performance liquid chromatography.The antioxidant activity of dihydromyricetin and vine tea’s flavonoid-rich extract was evaluated in vitro using six different methods.Results:Vine tea contained a large number of flavonoids,with dihydromyricetin as its main constituent.The flavonoid-rich extract exhibited a significant scavenging effect on superoxide anion radicals,and on 3-ethylbenzthiazoline-6-sulphonic acid and 1,1-diphenyl-2-picrylhydrazyl radicals.It also possessed definite activity in lipid peroxidation inhibition,ferric reduction,and the moderation of Fe2+ion chelation ability.There was a significant negative correlation between dihydromyricetin content and antioxidant activity in the vine tea samples,including superoxide anion radical scavenging activity(P=−0.754,P<0.05),lipid peroxidation inhibition activity(P=−0.759,P<0.05),ferric-reducing antioxidant power(P=−0.843,P<0.01),respectively.Dihydromyricetin played a dominant role in the antioxidant activities of the flavonoid-rich extract.Conclusion:Vine tea’s flavonoid-rich extract could be used as a new antioxidant source to safeguard against oxidative stress.展开更多
Dry powder inhaler Liposomes were prepared to investigate the effectiveness of pulmonary delivery of Colchicine and Budesonide for Idiopathic Pulmonary fibrosis. Budesonide(BUD) and Colchicine(COL) liposomes were prep...Dry powder inhaler Liposomes were prepared to investigate the effectiveness of pulmonary delivery of Colchicine and Budesonide for Idiopathic Pulmonary fibrosis. Budesonide(BUD) and Colchicine(COL) liposomes were prepared by thin layer film hydration method(TFH) using 1,2-Dipalmitoyl-sn-glycero-3-phosphoglycerol sodium(DPPG), Hydrogenated Soyaphosphotidylcholine(HSPC), Soyaphosphatidylcholine(SPC), cholesterol(CHOL) and drug in different weight ratios. The optimum lipid composition for BUD(74.22 ± 0.97%) was DPPG:HSPC: CHOL(4:5:1) and for COL(50.94 ± 2.04%) was DPPG: SPC: CHOL(3:6:1). These compositions retained drug for a longer period of time so selected for further study. Liposomes were found to be spherical in shape with mean size below 100 nm. Liposomes lyophilized using Mannitol as carrier and cryoprotectant showed high entrapment efficiency(97.89-98.6%). The powder was dispersed through an Andersen cascade impactor to evaluate the performance of the aerosolized powder. It was found that prepared liposomal dry powder inhaler(DPIs) sustained the drug release up to 24 hours. Optimized Budesonide DPI Formulation B2(86.53 ± 1.9%), Colchicine DPI Formulation C2(90.54 ± 2.3 %) and BUD and COL DPI Combination M2(89.91 ± 1.8%, 91.23 ± 1.9%). Histopathological results, measurements of lung hydroxyproline content, Myeloperoxidase activity indicated that liposomal drypowder inhaler administration attenuates lung fibrosis induced by bleomycin. Long term stability studies indicated that lyophilised BUD and COL liposomes were stable for 6 months at(25 °C± 2 °C, 60% ± 5% RH) and refrigerated conditions(2-8 °C). These results supported that combination of budesonide and colchicine liposomal dry powder inhaler pulmonary drug delivery for treatment of idiopathic Pulmonary Fibrosis exhibits prolonged drug retention at targeted site and reduces the systemic exposure.展开更多
The aim of the present study was to design and evaluate single pulse and floating double pulse valsartan core-in-cup tablets.Core tablets were prepared by direct compression of a homogenous mixture of valsartan,Avice...The aim of the present study was to design and evaluate single pulse and floating double pulse valsartan core-in-cup tablets.Core tablets were prepared by direct compression of a homogenous mixture of valsartan,Avicel PH-101,Croscarmellose sodium(CCNa),magnesium stearate&Aerosil.Weight variation,Hardness and Disintegration time were measured for the core tablets.Core-in-cup tablets were formulated using different polymers as a plug layer,including sodium alginate(SA),sodium carboxymethylcellulose(NaCMC)and hydroxypropyl methyl cellulose(HPMC).The floating behavior,water uptake and drug release from the prepared formulations were evaluated.Differential Scanning Calorimetry(DSC)was also performed to detect the possible drug excipient interaction.Stability study of the selected formula was performed at 25C&60%RH and at 40C&75%RH for 3 months.Finally,the existence of the selected formula in the stomach after oral administration to human volunteers was verified via x-ray radiography.The results showed that the release lag time of the tablets increased when the quantity of the plug layer increased thus decreasing the drug release.Plug layer polymers showed a lag time with rank order:SA<NaCMC<HPMC.Selected formulations are F5&F6.F5(having SA as the plug polymer)released valsartan after a lag time of 2 h while F6 released the drug in two successive pulses with a reasonable lag time in between due to its floating behavior.Formulations were stable for at least 3 months under standard long-term and accelerated storage conditions.In conclusion,pulsatile single pulse and floating double pulse stable valsartan core-incup tablets were successfully formulated which provided a desirable lag time followed by a rapid drug release.展开更多
Objective To investigate the genotypic diversity of Methicillin-resistant Staphylococcus aureus(MRSA) isolated from pigs and retail foods from different geographical areas in China and further to study the routes and ...Objective To investigate the genotypic diversity of Methicillin-resistant Staphylococcus aureus(MRSA) isolated from pigs and retail foods from different geographical areas in China and further to study the routes and rates of transmission of this pathogen from animals to food. Methods Seventy-one MRSA isolates were obtained from pigs and retail foods and then characterized by multi-locus sequencing typing(MLST), spa typing, multiple-locus variable number of tandem repeat analysis(MLVA), pulsed-field gel electrophoresis(PFGE), and antimicrobial susceptibility testing. Results All isolated MRSA exhibited multi-drug resistance(MDR). Greater diversity was found in food-associated MRSA(7 STs, 8 spa types, and 10 MLVA patterns) compared to pig-associated MRSA(3 STs, 1 spa type, and 6 MLVA patterns). PFGE patterns were more diverse for pig-associated MRSA than those of food-associated isolates(40 vs. 11 pulse types). Among the pig-associated isolates, CC9-ST9-t899-MC2236 was the most prevalent clone(96.4%), and CC9-ST9-t437-MC621(20.0%) was the predominant clone among the food-associated isolates. The CC9-ST9 isolates showed significantly higher antimicrobial resistance than other clones. Interestingly, CC398-ST398-t034 clone was identified from both pig-and food-associated isolates. Of note, some community-and hospital-associated MRSA strains(t030, t172, t1244, and t4549) were also identified as food-associated isolates. Conclusion CC9-ST9-t899-MC2236-MDR was the most predominant clone in pigs, but significant genetic diversity was observed in food-associated MRSA. Our results demonstrate the great need for improved surveillance of MRSA in livestock and food and effective prevention strategies to limit MDR-MRSA infections in China.展开更多
Repositioning of dipeptidyl peptidase-4 inhibitors and glucagon like peptide-1 receptor agonists is a breakthrough in the field of neural regeneration research increasing glucagon like peptide-1 bioavailability, hence...Repositioning of dipeptidyl peptidase-4 inhibitors and glucagon like peptide-1 receptor agonists is a breakthrough in the field of neural regeneration research increasing glucagon like peptide-1 bioavailability, hence its neuroprotective activities. In this article, the authors suggest not only crossing blood-brain barrier and neurodegenerative disease as off target for dipeptidyl peptidase-4 inhibitors and glucagon like peptide-1 receptor agonists, but also for ophthalmic preparations for diabetic retinopathy, which may be the latest breakthrough in the field if prepared and used in an appropriate nano-formulation to target the retinal nerves. The relation of neurodegenerative diseases' different mechanisms to the dipeptidyl peptidase-4 inhibitors and glucagon like peptide-1 receptor agonists should be further examined in preclinical and clinical settings. The repositioning of already marketed antidiabetic drugs for neurodegenerative diseases should save the high cost of the time-consuming normal drug development process. Drug repositioning is a hot topic as an alternative to molecular target based drug discovery or therapeutic switching. It is a relatively inexpensive pathway due to availability of previous pharmacological and safety data. The glucagon like peptide-1 produced in brain has been linked to enhanced learning and memory functions as a physiologic regulator in central nervous system by restoring insulin signaling. Intranasal administration of all marketed gliptins(or glucagon like peptide-1 receptor agonists) may show enhanced blood-brain barrier crossing and increased glucagon like peptide-1 levels in the brain after direct crossing of the drug for the olfactory region, targeting the cerebrospinal fluid. Further blood-brain barrier crossing tests may extend dipeptidyl peptidase-4 inhibitors' effects beyond the anti-hyperglycemic control to intranasal spray, intranasal powder, or drops targeting the blood-brain barrier and neurodegenerative diseases with the most suitable formula. Moreover, novel nano-formulation is encouraged either to obtain favorable pharmacokinetic parameters or to achieve promising blood-brain barrier penetration directly through the olfactory region. Many surfactants should be investigated either as a solubilizing agent for hydrophobic drugs or as penetration enhancers. Different formulae based on in vitro and in vivo characterizations, working on sister gliptins(or glucagon like peptide-1 receptor agonists), different routes of administration, pharmacokinetic studies, dose response relationship studies, monitoring of plasma/brain concentration ratio after single and multiple dose, and neurodegenerative disease animal models are required to prove the new method of use(utility) for dipeptidyl peptidase-4 inhibitors as potential neuroprotective agents. Furthermore, investigations of glucagon like peptide-1 receptor agonists' neuroprotective effects on animal models will be considered carefully because they crossed the blood-brain barrier in previous studies, enabling their direct action on the central nervous system. Combination therapy of dipeptidyl peptidase-4 inhibitors or glucagon like peptide-1 receptor agonists with already marketed drugs for neurodegenerative disease should be considered, especially regarding the novel intranasal route of administration.展开更多
The World Health Organization(WHO)declared global SARS-CoV-2(COVID-19)pandemic status on March 11,2020[1].To date,data record approximately 106 million infected individuals and 2.32 million deaths due to pandemic COVI...The World Health Organization(WHO)declared global SARS-CoV-2(COVID-19)pandemic status on March 11,2020[1].To date,data record approximately 106 million infected individuals and 2.32 million deaths due to pandemic COVID-19,worldwide.The most common symptoms of SARS-CoV-2 infection reported are:elevation of body temperature,fatigue,cough,loss of smell.In a percentage of patients,especially elderly individuals with comorbidities,COVID-19 infection can cause severe organ injury[2].Since the onset of the COVID-19 pandemic,numerous pharmacological treatments have been used off label,to treat the viral infection,with the primary aim of avoiding the most serious complications and organ injury.展开更多
AIM:To explore that if peroxynitrite induced the expression of inducible nitric oxide synthase(iNOS)via nuclear factor-kappa B(NF-κB)pathway in retinal pigment epithelial(RPE) cells and the antagonism of cholecystoki...AIM:To explore that if peroxynitrite induced the expression of inducible nitric oxide synthase(iNOS)via nuclear factor-kappa B(NF-κB)pathway in retinal pigment epithelial(RPE) cells and the antagonism of cholecystokinin octapeptide-8(Melatonin,CCK-8) in vitro.METHODS:RPE cells were obtained from eyes of C57BL/6 mouse and divided into control,peroxynitrite and CCK-8 groups.Control group was treated with saline,peroxynitrite group was treated with peroxynitrite,and CCK-8 group was treated with CCK-8 after added with peroxynitrite.All changes were observered at 6,12 and 24 hours after treatment.Gene array analysis,Reverse Transcription Polymerase Chain Reaction(RT-PCR) were used to determine the expression of inducible nitric oxide synthase(iNOS)mRNA in RPE cells.Western blotting was used to test the apoptosis of RPE cells.Immunofluorescent staining was used to determine the NF-κB pathway signal transduction.RESULTS:Compared to the control group,the expression of iNOS mRNA was up-regulated in peroxynitrite group and down-regulated in CCK-8 group with gene array analysis.Apoptosis was increased in peroxynitrite group and decreased in CCK-8 group with western blotting.The NF-κB pathway signal transduction was more and more stronger in the peroxynitrite group.But in CCK-8 group,little stronger could be observed at 12 hours,then weak at 24 hours with immunofluorescent staining(P <0.001).CONCLUSION:This study suggested that apoptosis of RPE cells was partly induced by peroxynitrite,which may be the new way of oxidative damage to the RPE cells.The NF-κB signal transduction may affect and reinforce apoptosis mediated by peroxynitrite.CCK-8 decreased apoptosis of RPE cells induced by peroxynitrite and is a potential agent for therapy of retinopathy.The mechanism of CCK-8 dealing with RPE cells may be related to its direct inhibition of the formation of iNOS to produce peroxynitrite and antagnism of damage of peroxynitrite to the RPE cells.展开更多
Objective:To explore the in vivo anticancer,anti-angiogenesis and immunomodulatory efficacies of the bioactive polysaccharide isolated from cold aqueous extract of Jania rubens(JCEM) and Pterocladia capillacea(PCEM) a...Objective:To explore the in vivo anticancer,anti-angiogenesis and immunomodulatory efficacies of the bioactive polysaccharide isolated from cold aqueous extract of Jania rubens(JCEM) and Pterocladia capillacea(PCEM) as well as hot aqueous extract of Enteromorpha intestinalis(EHEM) against hepatocellular carcinoma rat model(HCC) and to study their chemical composition.Methods:The sugars and amino acids composition of the bioactive polysaccharides of JCEM,PCEM and EHEM were determined using gas liquid chromatography and amino acid analyzer,respectively.These polysaccharide extracts(20 mg/kg b.wt.for 5 weeks) were assessed on hepatocarcinogenesis in rats and α-fetoprotein(AFP),carcinoembryonic antigen(CEA),glypican-3(GPC-3),hepatocyte growth factor(HGF) and vascular endothelial growth factor(VEGF) and Ig G levels were evaluated.Results:The GLC analysis of JCEM,PCEM and EHEM polysaccharide revealed the presence of 10,9 and10 sugars,in addition the amino acid analyser enable identification of 16,15 and 15 amino acids,respectively.These polysaccharide extracts of JCEM,PCEM and EHEM produced significant decrease in serum AFP,CEA,GPC-3,HGF and VEGF compared with untreated HCC group.JCEM,PCEM and EHEM had an immunostimulatory responses by increasing the IgG levels as compared by naive value(1.23,1.53 and 1.17 folds),respectively.The bioactive polysaccharides in HCC induced rats improved the humoral immune response.The photomicrographs of liver tissue sections of the groups of HCC treated with polysaccharide extracts of Jania rubens and Enteromorpha intestinalis showed intact histological structure.Moreover,fractions HE1,HE4,HE7 obtained from polysaccharide of EHEM showed moderate cytotoxic activity against Hep G2 in vitro with IC_(50) 73.1,42.6,76.2 μg/mL.However,fractions of PCEM and JCEM show no or weak cytotoxicity against Hep G2 in vitro where the cytotoxic activity of their crude polysaccharide extract proved synergetic effect.Conclusions:The pronounced antitumor activity of sulphated polysaccharide-protein complexes of JCEM and EHEM is due to direct cytotoxic activity,anti-hepatocarcinogensis,and anti-angiogenesis.In addition,JCEM,PCEM and EHEM had an immunostimulatory response and improved the humoral immune response in HCC induced rats.展开更多
Fungal keratitis and endopthalmitis are serious eye diseases.Fluconazole(FL)is indicated for their treatment,but suffers from poor topical ocular availability.This study was intended to improve and prolong its ocular ...Fungal keratitis and endopthalmitis are serious eye diseases.Fluconazole(FL)is indicated for their treatment,but suffers from poor topical ocular availability.This study was intended to improve and prolong its ocular availability.FL niosomal vesicles were prepared using span 60.Also,polymeric nanoparticles were prepared using cationic Eudragit RS100 and Eudragit RL100.The investigated particles had adequate entrapment efficiency(EE%),nanoscale particle size and high zeta potential.Subsequently,formulations were optimized using full factorial design.FL-HP-β-CD complex was encapsulated in selected Eudragit nanoprticles(FL-CD-ERS1)and niosmal vesicles.The niosomes were further coated with cationic and bioadhesive chitosan(FL-CD-Nios-ch).EE%for FL-CD-ERS1 and FL-CD-Niosch formulations were 76.4%and 61.7%;particle sizes were 151.1 and 392 nm;also,they exhibited satisfactory zeta potential+40.1 and+28.5 m V.In situ gels were prepared by poloxamer P407,HPMC and chitosan and evaluated for gelling capacity,rheological behavior and gelling temperature.To increase the precorneal residence time,free drug and selected nano-formulations were incorporated in the selected in situ gel.Release study revealed sustained release within 24 h.Permeation through excised rabbits corneas demonstrated enhanced drug flux and large AUC0-6 h in comparison to plain drug.Corneal permeation of selected formulations labeled with Rhodamine B was visualized by Confocal laser microscopy.Histopathological study and in vivo tolerance test evidenced safety.In vivo susceptibility test using Candida albicans depicted enhanced growth inhibition and sustained effect.In this study the adopted stepwise optimization strategy combined cylodextrin complexation,drug nano-encapsulation and loading within thermosenstive in situ gel.Finally,the developed innovated formulations displayed boosted corneal permeation,enhanced antifungal activity and prolonged action.展开更多
Tadalafil(TAD)and dapoxetine HCl(DAP)are recently co-formulated and both show native fluorescence.Therefore,a novel,accurate,specific and sensitive reversed-phase high performance liquid chromatographic method with fl...Tadalafil(TAD)and dapoxetine HCl(DAP)are recently co-formulated and both show native fluorescence.Therefore,a novel,accurate,specific and sensitive reversed-phase high performance liquid chromatographic method with fluorescence detection was developed and validated for their separation and quantitation in dosage form and human plasma using avanafil as an internal standard(IS).Separation was achieved using isocratic elution within 7.0 min on C18column and acetonitrile-0.15%triethylamine(40∶60,v/v;pH 4)as a mobile phase.The flow rate was 1.0 mL/min and the detection was time-programmed at 330,410 and 370 nm for TAD,DAP and IS,respectively,after excitation at 236 nm.The linear ranges from 0.01 to 30.00μg/mL for each drug with the limits of detection of 4.20 and 7.20 ng/mL for TAD and DAP,respectively.The method was validated in accordance to the International Conference on Harmonization(ICH)guidelines and was successfully applied to spiked human plasma with mean recoveries of 98.17%and 98.83%for TAD and DAP respectively.展开更多
AIM: To investigate the relationship of changes in expression of marker genes in functional categories or molecular networks comprising one functional category or multiple categories in progression of hepatic fibrosis...AIM: To investigate the relationship of changes in expression of marker genes in functional categories or molecular networks comprising one functional category or multiple categories in progression of hepatic fibrosis in hepatitis C (HCV) patients. METHODS: Marker genes were initially identified using DNA microarray data from a rat liver fibrosis model. The expression level of each fibrosis associated marker gene was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) in clinical biopsy specimens from HCV-positive patients (n = 61). Analysis of changes in expression patterns and interactions of marker genes in functional categories was used to assess the biological mechanism of fibrosis. RESULTS: The profile data showed several biological changes associated with progression of hepatic fibrosis. Clustered genes in functional categories showed sequential changes in expression. Several sets of clustered genes, including those related to the extracellular matrix (ECM), inflammation, lipid metabolism, steroid metabolism, and some transcription factors important for hepatic biology showed expression changes in the immediate early phase (F1/F2) of fibrosis. Genes associated with aromatic amino acid (AA) metabolism, sulfur-containing AA metabolism and insulin/ Wnt signaling showed expression changes in the middle phase (F2/F3), and some genes related to glucose metabolism showed altered expression in the late phase of fibrosis (F3/F4). Therefore, molecular networks showing serial changes in gene expression are present in liver fibrosis progression in hepatitis C patients. CONCLUSION: Analysis of gene expression profiles from a perspective of functional categories or molecular networks provides an understanding of disease and suggests new diagnostic methods. Selected marker genes have potential utility for biological identification of advanced fibrosis.展开更多
A rat model of diabetes mellitus was established by intraperitoneal injection of streptozotocin. Three days later, the rats were intraperitoneally administered 140 mg puerarin/kg daily, for a total of 60 successive da...A rat model of diabetes mellitus was established by intraperitoneal injection of streptozotocin. Three days later, the rats were intraperitoneally administered 140 mg puerarin/kg daily, for a total of 60 successive days. DNA ladder results showed increased apoptosis over time in retinal pigment epithelial cells from rats with streptozotocin-induced diabetes mellitus. Western blot analysis, Reverse transcription-PCR, immunohistochemistry, and flow cytometry results showed increased expression of 3-nitrotyrosine, a peroxyntrite marker, as well as inducible nitric synthase and Fas/FasL, in retinal pigment epithelial cells. Puerarin reversed these changes, and results demonstrated that puerarin inhibited Fas/FasL expression and alleviated peroxyntrite injury to retinal pigment epithelial cells. These results suggested that puerarin inhibited production of inducible nitric oxide synthase and directly antagonized peroxyntrite injury in retinal pigment epithelial cells.展开更多
BACKGROUND Anti-programmed death therapy has thrust immunotherapy into the spotlight.However,such therapy has a modest response in hepatocellular carcinoma(HCC).Epigenetic immunomodulation is a suggestive combinatoria...BACKGROUND Anti-programmed death therapy has thrust immunotherapy into the spotlight.However,such therapy has a modest response in hepatocellular carcinoma(HCC).Epigenetic immunomodulation is a suggestive combinatorial therapy with immune checkpoint blockade.Non-coding ribonucleic acid(ncRNA)driven regulation is a major mechanism of epigenetic modulation.Given the wide range of ncRNAs that co-opt in programmed cell-death protein 1(PD-1)/programmed death ligand 1(PD-L1)regulation,and based on the literature,we hypothesized that miR-155-5p,miR-194-5p and long non-coding RNAs(lncRNAs)X-inactive specific transcript(XIST)and MALAT-1 are involved in a regulatory upstream pathway for PD-1/PD-L1.Recently,nutraceutical therapeutics in cancers have received increasing attention.Thus,it is interesting to study the impact of oleuropein on the respective study key players.AIM To explore potential upstream regulatory ncRNAs for the immune checkpoint PD-1/PD-L1.METHODS Bioinformatics tools including microrna.org and lnCeDB software were adopted to detect targeting of miR-155-5p,miR-194-5p and lncRNAs XIST and MALAT-1 to PD-L1 mRNA,respectively.In addition,Diana tool was used to predict targeting of both aforementioned miRNAs to lncRNAs XIST and MALAT-1.HCC and normal tissue samples were collected for scanning of PD-L1,XIST and MALAT-1 expression.To study the interaction among miR-155-5p,miR-194-5p,lncRNAs XIST and MALAT-1,as well as PD-L1 mRNA,a series of transfections of the Huh-7 cell line was carried out.RESULTS Bioinformatics software predicted that miR-155-5p and miR-194-5p can target PDL1,MALAT-1 and XIST.MALAT-1 and XIST were predicted to target PD-L1 mRNA.PD-L1 and XIST were significantly upregulated in 23 HCC biopsies compared to healthy controls;however,MALAT-1 was barely detected.MiR-194 induced expression elevated the expression of PD-L1,XIST and MALAT-1.However,overexpression of miR-155-5p induced the upregulation of PD-L1 and XIST,while it had a negative impact on MALAT-1 expression.Knockdown of XIST did have an impact on PD-L1 expression;however,following knockdown of the negative regulator of X-inactive specific transcript(TSIX),PD-L1 expression was elevated,and abolished MALAT-1 activity.Upon co-transfection of miR-194-5p with siMALAT-1,PD-L1 expression was elevated.Co-transfection of miR-194-5p with siXIST did not have an impact on PD-L1 expression.Upon co-transfection of miR-194 with siTSIX,PD-L1 expression was upregulated.Interestingly,the same PD-L1 expression pattern was observed following miR-155-5p cotransfections.Oleuropein treatment of Huh-7 cells reduced the expression profile of PD-L1,XIST,and miR-155-5p,upregulated the expression of miR-194-5p and had no significant impact on the MALAT-1 expression profile.CONCLUSION This study reported a novel finding revealing that opposing acting miRNAs in HCC,have the same impact on PD-1/PD-L1 immune checkpoint by sharing a common signaling pathway.展开更多
AIM: To determine the gene expression profile data for the whole liver during development of dimethylni-trosamine (DMN)-induced hepatic fibrosis.METHODS: Marker genes were identified for different types of hepatic cel...AIM: To determine the gene expression profile data for the whole liver during development of dimethylni-trosamine (DMN)-induced hepatic fibrosis.METHODS: Marker genes were identified for different types of hepatic cells, including hepatic stellate cells (HSCs), Kupffer cells (including other inflammatory cells), and hepatocytes, using independent temporal DNA microarray data obtained from isolated hepatic cells. RESULTS: The cell-type analysis of gene expression gave several key results and led to formation of three hypotheses: (1) changes in the expression of HSC-specific marker genes during fibrosis were similar to gene expression data in in vitro cultured HSCs, suggesting a major role of the self-activating characteristics of HSCs in formation of fibrosis; (2) expression of mast cell-specific marker genes reached a peak during liver fibrosis, suggesting a possible role of mast cells in formation of fibrosis; and (3) abnormal expression of hepatocyte-specific marker genes was found across several metabolic pathways during fibrosis, including sulfur-containing amino acid metabolism, fatty acid metabolism, and drug metabolism, suggesting a mechanistic relationship between these abnormalities and symptoms of liver fibrosis. CONCLUSION: Analysis of marker genes for specific hepatic cell types can identify the key aspects of fibro-genesis. Sequential activation of inflammatory cells and the self-supporting properties of HSCs play an important role in development of fibrosis.展开更多
Objective:To evaluate the scolicidal and immunomodulatory effect of the Ziziphora tenuior(Z.tenuior) extract and its fractions.Methods:Protoscolices were treated with six concentrations(3,5,10,25,50,and 100 mg/mL) of ...Objective:To evaluate the scolicidal and immunomodulatory effect of the Ziziphora tenuior(Z.tenuior) extract and its fractions.Methods:Protoscolices were treated with six concentrations(3,5,10,25,50,and 100 mg/mL) of Z.tenuior extract and its fractions(ethanol,petroleum ether,ethyl acetate and chloroform) in periods of 10,20,30,40,50 and 60 minutes,and viability of protoscolices was evaluated using the 1.0%eosin.To examine the immunomodulatory effects of Ziziphora and its fractions on macrophage cells,the non-toxic concentration of extract and different fractions determined by MTT assay,and the Griess reaction was used to measure the level of nitrite as an indicator of nitric oxide by the macrophage cells in 10,100 and 200 μg/mL in 24 hours at 37 ℃.Results:In this study,the Z.tenuior extract at 10 mg/mL concentration was able to kill all protoscolices during 20 minutes.By increasing the concentration to 25 mg/mL,the scolicidal time reduced to 10 minutes.Regarding the effect of different fractions of Z.tenuior,the ethanolic fraction showed the highest scolicidal activity.The extract demonstrated an inhibitory effect on the activity of macrophages and reduced nitric oxide production.Although the petroleum ether and ethanolic fractions of the extract reduced nitric oxide production,nevertheless,this effect was only significant at 10 and 100 ug/mL concentrations(P<0.05).Conclusion:The Z.tenuior extract and its fractions were effective against protoscolices yet the effect of total extract was considerable.Our findings indicates that the extract and its ethanolic and petroleum ether fractions could have anti-inflammatory properties.展开更多
文摘Tobramycin is one of the aminoglycoside antibiotics that lack a UV absorbing chromophore. However, the application of pulsed electrochemical detection (PED) has been used successfully for the analysis of this and similar antibiotics. This work describes an improved liquid chromatographic (LC) method combined with PED, which is able to separate much more impurities than before. Using a Discovery C-18 RP column (250 mm 4.6 mm i.d., 5 mm), isocratic elution was carried out with a mobile phase, containing sodium sulfate (35 g/L), sodium octanesulphonic acid (1 g/L), tetrahydrofuran (14 mL/L) and 0.2 M phosphate buffer pH 3.0 (50 mL/L). Using these experimental conditions, the limit of quantification (LOQ, S/N 10) was 5 ng. The linearity was examined in the range LOQ-60 mg/mL and the coefficient of determination was 0.998. The method also proved to be repeatable and the recovery was close to 100%. The influence of the different chromatographic parameters on the separation was investigated by means of an experimental design. The proposed method is useful in quality control of tobramycin drug substances and drug products.
文摘Cefuroxime Axetil (CA) a widely used cephalosporin antibiotic displays low aqueous solubility and high membrane penetrability. This results in its solubility driven variable and/or low oral bioavailability and therapeutic efficacy as a major drawback. Thus, most of the goal of our study was to increase the solubility as well as dissolution rate of CA using the simple and cost-effective solid dispersion (SD) method. At first, the SD formulations of CA were prepared at various weight ratios of Carplex-67 and PEG-4000 by solvent evaporation technique. These new formulations were then subjected to an in-vitro drug release performance study and tested for physicochemical characterization to distinguish the thermal behavior, crystallinity, interactions phenomena, and surface morphology. Among the formulated Cefuroxime Axetil Solid Dispersion (CSD), CSD-8 which contained CA, Carplex-67, and PEG-4000 at the weight ratio 1:3:2, respectively showed the most significant (p in-vitro dissolution in water, Gastric Simulated Fluid (GSF), and Intestinal Simulated Fluid (ISF). This study also showed a significant (p < 0.001) increase in drug release compared to the marketed product. Therefore, it is supposed to be a promising alternative to conventional antimicrobial therapy.
基金supported by the National Key R&D Program of China (2018YFC1706506)the National Natural Science Foundation of China Youth Program (81703921)the Tianjin Key Medical Discipline (Specialty)Construction Project。
文摘Objective:Liver cirrhosis is a disease that seriously damages human health.Traditional Chinese Medicine(TCM)formulae have a good therapeutic effect on cirrhosis,and the herb pair is the smallest unit in formula compatibility,which is important for improving the therapeutic effect.Therefore,identifying core herb pairs among TCM formulae is key.Methods:We mined the data of TCM formulae for the treatment of cirrhosis in the China National Intellectual Property Administration for the first time and analyzed their herb characteristics and association rules.We screened 405 patented TCM formulae,including 953 herbs.Based on frequency statistics and association rules,we determined“Astragali Radix-Salviae Miltiorrhizae Radix Et Rhizoma”as the core herb pair.Results:Six active compounds,Isorhamnetin,Formononetin,Calycosin,Cryptotanshinone,Dihydrotanshinone I,and Tanshinone II A,were screened out based on previous studies and network pharmacology.We found that SRC,TP53,HSP90AA1,MAPK3,MAPK1,and STAT3 played pivotal roles in treating cirrhosis.Interestingly,molecular docking indicated that MAPK3 might be a potential pharmacological target for cirrhosis.Conclusion:We preliminarily predicted and verified the pharmacological and molecular mechanism of“Astragali Radix-Salviae Miltiorrhizae Radix Et Rhizoma”in treating cirrhosis.This can expand the scope of TCM in the treatment of cirrhosis,guide people to use clinical formulae,and provide valuable insights for further drug discovery studies.
文摘Paracetamol (PCM) is enlisted in the WHO model list as an essential medicine for pain and palliative care, but at overdose, it causes hepatic damage. This study was designed to assess the analgesic efficacy and hepatoprotective property of a solid dispersion (SD) loaded with PCM. A number of PCM loaded formulations (PSDs) were fabricated using silica alone or in combination with polyethylene glycol and/or Na-citrate followed by in-vitro dissolution profiling. Selected PSDs with improved dissolution profile were subjected to solid-state characterization (DSC, PXRD, FTIR, and SEM), stability study along with investigation of in-vivo analgesic efficacy and effect on hepatocytes. Among these, PSD10 showed a rapid and significantly higher in-vitro drug release than pure PCM. This improvement was distinct to other PSDs also. Solid-state characterization of PSD10 authenticated the conversion of crystalline PCM to amorphous form upon formulation. Subsequent oral administration of PSD10 in Swiss albino mice showed 1.44-fold greater analgesic efficacy than pure PCM at dose 30 mg/kg. Besides, at acute toxic dose, liver histology of PSD10 mice was comparable with NC mice indicating hepatic protection upon formulation, whereas the PCM mice showed extensive hepatic necrosis which was also endorsed by significantly higher values of SGPT, SGOT, and ALP than PSD10 mice. Finally, an accelerated stability study of PSD10 performed according to the guideline of ICH noticed no remarkable deviation in its dissolution performance as well as crystalline nature. Thus, this newly developed PSD10 may be a safe and promising alternative for pain management and palliative care.
基金supported by Health Department of Shandong Province,Science and Technology Program Fund Assistance ltem(No:2010HZ102)
文摘Objective:To observe the effect of co-culture cytokine-induced killer cells(CIK) and homologous dendritic cells(DC) on the proliferative activity and phenotype change of the DCCIK cell and the cell killing activity of leukemia HL-60.Methods:50 mL cord blood sample was obtained from infants delivered by full term healthy woman and the cord blood mononuclear cells were isolated by density gradient centrifugal ion.Non-adherent cells were collectedfor the induction culture of CIK.adherent cells were differentiated into mature DC;cultured mature DC was mixed with and CIK in the proportion of 1:5 for 12 d.killing activity of DC-CIK co-cultured cell on leukemia HL-60 was detected by MTT assay.Results:Compared with CIKs.the cocultured DC-CIKs presented a markedly higher proliferation and killing activity.Conclusions:Co-culture of DC-CIK cells led to a significant increase of the proliferation and cytotoxicity of CIK.
基金This work was supported by Traditional Chinese Medicine Science and Technology Development Project of Shandong Province(No.2019-0649)Fundamental Research Funds for the Central Universities(No.2018-BUCMXJKY001).
文摘Background:Vine tea from fermented Ampelopsis grossedentata leaves has been used as a herbal tea and folk medicine in the southern region of China for hundreds of years.The aim of this investigation was to analyze the total flavonoids found in vine tea,including three bioactive flavonoids,and the total phenolic contents in the aqueous methanol extracts of 10 vine tea samples.In addition,this study also aimed to examine the antioxidant activity of dihydromyricetin and vine tea’s flavonoid-rich extract.Methods:The total flavonoids and total phenolic content assay of extracts from vine tea were performed by ultraviolet-visible spectroscopy and epoch microplate spectrophotometer,respectively.Three bioactive flavonoids were quantified simultaneously using high performance liquid chromatography.The antioxidant activity of dihydromyricetin and vine tea’s flavonoid-rich extract was evaluated in vitro using six different methods.Results:Vine tea contained a large number of flavonoids,with dihydromyricetin as its main constituent.The flavonoid-rich extract exhibited a significant scavenging effect on superoxide anion radicals,and on 3-ethylbenzthiazoline-6-sulphonic acid and 1,1-diphenyl-2-picrylhydrazyl radicals.It also possessed definite activity in lipid peroxidation inhibition,ferric reduction,and the moderation of Fe2+ion chelation ability.There was a significant negative correlation between dihydromyricetin content and antioxidant activity in the vine tea samples,including superoxide anion radical scavenging activity(P=−0.754,P<0.05),lipid peroxidation inhibition activity(P=−0.759,P<0.05),ferric-reducing antioxidant power(P=−0.843,P<0.01),respectively.Dihydromyricetin played a dominant role in the antioxidant activities of the flavonoid-rich extract.Conclusion:Vine tea’s flavonoid-rich extract could be used as a new antioxidant source to safeguard against oxidative stress.
文摘Dry powder inhaler Liposomes were prepared to investigate the effectiveness of pulmonary delivery of Colchicine and Budesonide for Idiopathic Pulmonary fibrosis. Budesonide(BUD) and Colchicine(COL) liposomes were prepared by thin layer film hydration method(TFH) using 1,2-Dipalmitoyl-sn-glycero-3-phosphoglycerol sodium(DPPG), Hydrogenated Soyaphosphotidylcholine(HSPC), Soyaphosphatidylcholine(SPC), cholesterol(CHOL) and drug in different weight ratios. The optimum lipid composition for BUD(74.22 ± 0.97%) was DPPG:HSPC: CHOL(4:5:1) and for COL(50.94 ± 2.04%) was DPPG: SPC: CHOL(3:6:1). These compositions retained drug for a longer period of time so selected for further study. Liposomes were found to be spherical in shape with mean size below 100 nm. Liposomes lyophilized using Mannitol as carrier and cryoprotectant showed high entrapment efficiency(97.89-98.6%). The powder was dispersed through an Andersen cascade impactor to evaluate the performance of the aerosolized powder. It was found that prepared liposomal dry powder inhaler(DPIs) sustained the drug release up to 24 hours. Optimized Budesonide DPI Formulation B2(86.53 ± 1.9%), Colchicine DPI Formulation C2(90.54 ± 2.3 %) and BUD and COL DPI Combination M2(89.91 ± 1.8%, 91.23 ± 1.9%). Histopathological results, measurements of lung hydroxyproline content, Myeloperoxidase activity indicated that liposomal drypowder inhaler administration attenuates lung fibrosis induced by bleomycin. Long term stability studies indicated that lyophilised BUD and COL liposomes were stable for 6 months at(25 °C± 2 °C, 60% ± 5% RH) and refrigerated conditions(2-8 °C). These results supported that combination of budesonide and colchicine liposomal dry powder inhaler pulmonary drug delivery for treatment of idiopathic Pulmonary Fibrosis exhibits prolonged drug retention at targeted site and reduces the systemic exposure.
基金Grateful thanks to Prof.Dr.Mohamed H.Zahran,Emeritus Professor of Radiology,Radiology Department,Faculty of Medicine,Alexandria University,for supervision of the X-ray study and his precious support.
文摘The aim of the present study was to design and evaluate single pulse and floating double pulse valsartan core-in-cup tablets.Core tablets were prepared by direct compression of a homogenous mixture of valsartan,Avicel PH-101,Croscarmellose sodium(CCNa),magnesium stearate&Aerosil.Weight variation,Hardness and Disintegration time were measured for the core tablets.Core-in-cup tablets were formulated using different polymers as a plug layer,including sodium alginate(SA),sodium carboxymethylcellulose(NaCMC)and hydroxypropyl methyl cellulose(HPMC).The floating behavior,water uptake and drug release from the prepared formulations were evaluated.Differential Scanning Calorimetry(DSC)was also performed to detect the possible drug excipient interaction.Stability study of the selected formula was performed at 25C&60%RH and at 40C&75%RH for 3 months.Finally,the existence of the selected formula in the stomach after oral administration to human volunteers was verified via x-ray radiography.The results showed that the release lag time of the tablets increased when the quantity of the plug layer increased thus decreasing the drug release.Plug layer polymers showed a lag time with rank order:SA<NaCMC<HPMC.Selected formulations are F5&F6.F5(having SA as the plug polymer)released valsartan after a lag time of 2 h while F6 released the drug in two successive pulses with a reasonable lag time in between due to its floating behavior.Formulations were stable for at least 3 months under standard long-term and accelerated storage conditions.In conclusion,pulsatile single pulse and floating double pulse stable valsartan core-incup tablets were successfully formulated which provided a desirable lag time followed by a rapid drug release.
基金funded by the National Key R&D Program of China(2016YFD0401102)(the National Key Research and Development Program of China,the Ministry of Science and Technology of China)China Food Safety Talent Competency Development Initiative:CFSA 523 Program(1311613106702)
文摘Objective To investigate the genotypic diversity of Methicillin-resistant Staphylococcus aureus(MRSA) isolated from pigs and retail foods from different geographical areas in China and further to study the routes and rates of transmission of this pathogen from animals to food. Methods Seventy-one MRSA isolates were obtained from pigs and retail foods and then characterized by multi-locus sequencing typing(MLST), spa typing, multiple-locus variable number of tandem repeat analysis(MLVA), pulsed-field gel electrophoresis(PFGE), and antimicrobial susceptibility testing. Results All isolated MRSA exhibited multi-drug resistance(MDR). Greater diversity was found in food-associated MRSA(7 STs, 8 spa types, and 10 MLVA patterns) compared to pig-associated MRSA(3 STs, 1 spa type, and 6 MLVA patterns). PFGE patterns were more diverse for pig-associated MRSA than those of food-associated isolates(40 vs. 11 pulse types). Among the pig-associated isolates, CC9-ST9-t899-MC2236 was the most prevalent clone(96.4%), and CC9-ST9-t437-MC621(20.0%) was the predominant clone among the food-associated isolates. The CC9-ST9 isolates showed significantly higher antimicrobial resistance than other clones. Interestingly, CC398-ST398-t034 clone was identified from both pig-and food-associated isolates. Of note, some community-and hospital-associated MRSA strains(t030, t172, t1244, and t4549) were also identified as food-associated isolates. Conclusion CC9-ST9-t899-MC2236-MDR was the most predominant clone in pigs, but significant genetic diversity was observed in food-associated MRSA. Our results demonstrate the great need for improved surveillance of MRSA in livestock and food and effective prevention strategies to limit MDR-MRSA infections in China.
文摘Repositioning of dipeptidyl peptidase-4 inhibitors and glucagon like peptide-1 receptor agonists is a breakthrough in the field of neural regeneration research increasing glucagon like peptide-1 bioavailability, hence its neuroprotective activities. In this article, the authors suggest not only crossing blood-brain barrier and neurodegenerative disease as off target for dipeptidyl peptidase-4 inhibitors and glucagon like peptide-1 receptor agonists, but also for ophthalmic preparations for diabetic retinopathy, which may be the latest breakthrough in the field if prepared and used in an appropriate nano-formulation to target the retinal nerves. The relation of neurodegenerative diseases' different mechanisms to the dipeptidyl peptidase-4 inhibitors and glucagon like peptide-1 receptor agonists should be further examined in preclinical and clinical settings. The repositioning of already marketed antidiabetic drugs for neurodegenerative diseases should save the high cost of the time-consuming normal drug development process. Drug repositioning is a hot topic as an alternative to molecular target based drug discovery or therapeutic switching. It is a relatively inexpensive pathway due to availability of previous pharmacological and safety data. The glucagon like peptide-1 produced in brain has been linked to enhanced learning and memory functions as a physiologic regulator in central nervous system by restoring insulin signaling. Intranasal administration of all marketed gliptins(or glucagon like peptide-1 receptor agonists) may show enhanced blood-brain barrier crossing and increased glucagon like peptide-1 levels in the brain after direct crossing of the drug for the olfactory region, targeting the cerebrospinal fluid. Further blood-brain barrier crossing tests may extend dipeptidyl peptidase-4 inhibitors' effects beyond the anti-hyperglycemic control to intranasal spray, intranasal powder, or drops targeting the blood-brain barrier and neurodegenerative diseases with the most suitable formula. Moreover, novel nano-formulation is encouraged either to obtain favorable pharmacokinetic parameters or to achieve promising blood-brain barrier penetration directly through the olfactory region. Many surfactants should be investigated either as a solubilizing agent for hydrophobic drugs or as penetration enhancers. Different formulae based on in vitro and in vivo characterizations, working on sister gliptins(or glucagon like peptide-1 receptor agonists), different routes of administration, pharmacokinetic studies, dose response relationship studies, monitoring of plasma/brain concentration ratio after single and multiple dose, and neurodegenerative disease animal models are required to prove the new method of use(utility) for dipeptidyl peptidase-4 inhibitors as potential neuroprotective agents. Furthermore, investigations of glucagon like peptide-1 receptor agonists' neuroprotective effects on animal models will be considered carefully because they crossed the blood-brain barrier in previous studies, enabling their direct action on the central nervous system. Combination therapy of dipeptidyl peptidase-4 inhibitors or glucagon like peptide-1 receptor agonists with already marketed drugs for neurodegenerative disease should be considered, especially regarding the novel intranasal route of administration.
文摘The World Health Organization(WHO)declared global SARS-CoV-2(COVID-19)pandemic status on March 11,2020[1].To date,data record approximately 106 million infected individuals and 2.32 million deaths due to pandemic COVID-19,worldwide.The most common symptoms of SARS-CoV-2 infection reported are:elevation of body temperature,fatigue,cough,loss of smell.In a percentage of patients,especially elderly individuals with comorbidities,COVID-19 infection can cause severe organ injury[2].Since the onset of the COVID-19 pandemic,numerous pharmacological treatments have been used off label,to treat the viral infection,with the primary aim of avoiding the most serious complications and organ injury.
基金Supported by Hebei Province Science foundation,China(No.07276101D-3)Clinical Science Project Fund of the Ministry of Health in Hebei Province,China(No. 03078)Foreign Studying Project Fund in Hebei Province,China
文摘AIM:To explore that if peroxynitrite induced the expression of inducible nitric oxide synthase(iNOS)via nuclear factor-kappa B(NF-κB)pathway in retinal pigment epithelial(RPE) cells and the antagonism of cholecystokinin octapeptide-8(Melatonin,CCK-8) in vitro.METHODS:RPE cells were obtained from eyes of C57BL/6 mouse and divided into control,peroxynitrite and CCK-8 groups.Control group was treated with saline,peroxynitrite group was treated with peroxynitrite,and CCK-8 group was treated with CCK-8 after added with peroxynitrite.All changes were observered at 6,12 and 24 hours after treatment.Gene array analysis,Reverse Transcription Polymerase Chain Reaction(RT-PCR) were used to determine the expression of inducible nitric oxide synthase(iNOS)mRNA in RPE cells.Western blotting was used to test the apoptosis of RPE cells.Immunofluorescent staining was used to determine the NF-κB pathway signal transduction.RESULTS:Compared to the control group,the expression of iNOS mRNA was up-regulated in peroxynitrite group and down-regulated in CCK-8 group with gene array analysis.Apoptosis was increased in peroxynitrite group and decreased in CCK-8 group with western blotting.The NF-κB pathway signal transduction was more and more stronger in the peroxynitrite group.But in CCK-8 group,little stronger could be observed at 12 hours,then weak at 24 hours with immunofluorescent staining(P <0.001).CONCLUSION:This study suggested that apoptosis of RPE cells was partly induced by peroxynitrite,which may be the new way of oxidative damage to the RPE cells.The NF-κB signal transduction may affect and reinforce apoptosis mediated by peroxynitrite.CCK-8 decreased apoptosis of RPE cells induced by peroxynitrite and is a potential agent for therapy of retinopathy.The mechanism of CCK-8 dealing with RPE cells may be related to its direct inhibition of the formation of iNOS to produce peroxynitrite and antagnism of damage of peroxynitrite to the RPE cells.
基金the National Research Centre for the financial support with Grant No.9080104
文摘Objective:To explore the in vivo anticancer,anti-angiogenesis and immunomodulatory efficacies of the bioactive polysaccharide isolated from cold aqueous extract of Jania rubens(JCEM) and Pterocladia capillacea(PCEM) as well as hot aqueous extract of Enteromorpha intestinalis(EHEM) against hepatocellular carcinoma rat model(HCC) and to study their chemical composition.Methods:The sugars and amino acids composition of the bioactive polysaccharides of JCEM,PCEM and EHEM were determined using gas liquid chromatography and amino acid analyzer,respectively.These polysaccharide extracts(20 mg/kg b.wt.for 5 weeks) were assessed on hepatocarcinogenesis in rats and α-fetoprotein(AFP),carcinoembryonic antigen(CEA),glypican-3(GPC-3),hepatocyte growth factor(HGF) and vascular endothelial growth factor(VEGF) and Ig G levels were evaluated.Results:The GLC analysis of JCEM,PCEM and EHEM polysaccharide revealed the presence of 10,9 and10 sugars,in addition the amino acid analyser enable identification of 16,15 and 15 amino acids,respectively.These polysaccharide extracts of JCEM,PCEM and EHEM produced significant decrease in serum AFP,CEA,GPC-3,HGF and VEGF compared with untreated HCC group.JCEM,PCEM and EHEM had an immunostimulatory responses by increasing the IgG levels as compared by naive value(1.23,1.53 and 1.17 folds),respectively.The bioactive polysaccharides in HCC induced rats improved the humoral immune response.The photomicrographs of liver tissue sections of the groups of HCC treated with polysaccharide extracts of Jania rubens and Enteromorpha intestinalis showed intact histological structure.Moreover,fractions HE1,HE4,HE7 obtained from polysaccharide of EHEM showed moderate cytotoxic activity against Hep G2 in vitro with IC_(50) 73.1,42.6,76.2 μg/mL.However,fractions of PCEM and JCEM show no or weak cytotoxicity against Hep G2 in vitro where the cytotoxic activity of their crude polysaccharide extract proved synergetic effect.Conclusions:The pronounced antitumor activity of sulphated polysaccharide-protein complexes of JCEM and EHEM is due to direct cytotoxic activity,anti-hepatocarcinogensis,and anti-angiogenesis.In addition,JCEM,PCEM and EHEM had an immunostimulatory response and improved the humoral immune response in HCC induced rats.
基金the National Research Centre,Cairo,Egypt for all the facilities and supports。
文摘Fungal keratitis and endopthalmitis are serious eye diseases.Fluconazole(FL)is indicated for their treatment,but suffers from poor topical ocular availability.This study was intended to improve and prolong its ocular availability.FL niosomal vesicles were prepared using span 60.Also,polymeric nanoparticles were prepared using cationic Eudragit RS100 and Eudragit RL100.The investigated particles had adequate entrapment efficiency(EE%),nanoscale particle size and high zeta potential.Subsequently,formulations were optimized using full factorial design.FL-HP-β-CD complex was encapsulated in selected Eudragit nanoprticles(FL-CD-ERS1)and niosmal vesicles.The niosomes were further coated with cationic and bioadhesive chitosan(FL-CD-Nios-ch).EE%for FL-CD-ERS1 and FL-CD-Niosch formulations were 76.4%and 61.7%;particle sizes were 151.1 and 392 nm;also,they exhibited satisfactory zeta potential+40.1 and+28.5 m V.In situ gels were prepared by poloxamer P407,HPMC and chitosan and evaluated for gelling capacity,rheological behavior and gelling temperature.To increase the precorneal residence time,free drug and selected nano-formulations were incorporated in the selected in situ gel.Release study revealed sustained release within 24 h.Permeation through excised rabbits corneas demonstrated enhanced drug flux and large AUC0-6 h in comparison to plain drug.Corneal permeation of selected formulations labeled with Rhodamine B was visualized by Confocal laser microscopy.Histopathological study and in vivo tolerance test evidenced safety.In vivo susceptibility test using Candida albicans depicted enhanced growth inhibition and sustained effect.In this study the adopted stepwise optimization strategy combined cylodextrin complexation,drug nano-encapsulation and loading within thermosenstive in situ gel.Finally,the developed innovated formulations displayed boosted corneal permeation,enhanced antifungal activity and prolonged action.
文摘Tadalafil(TAD)and dapoxetine HCl(DAP)are recently co-formulated and both show native fluorescence.Therefore,a novel,accurate,specific and sensitive reversed-phase high performance liquid chromatographic method with fluorescence detection was developed and validated for their separation and quantitation in dosage form and human plasma using avanafil as an internal standard(IS).Separation was achieved using isocratic elution within 7.0 min on C18column and acetonitrile-0.15%triethylamine(40∶60,v/v;pH 4)as a mobile phase.The flow rate was 1.0 mL/min and the detection was time-programmed at 330,410 and 370 nm for TAD,DAP and IS,respectively,after excitation at 236 nm.The linear ranges from 0.01 to 30.00μg/mL for each drug with the limits of detection of 4.20 and 7.20 ng/mL for TAD and DAP,respectively.The method was validated in accordance to the International Conference on Harmonization(ICH)guidelines and was successfully applied to spiked human plasma with mean recoveries of 98.17%and 98.83%for TAD and DAP respectively.
文摘AIM: To investigate the relationship of changes in expression of marker genes in functional categories or molecular networks comprising one functional category or multiple categories in progression of hepatic fibrosis in hepatitis C (HCV) patients. METHODS: Marker genes were initially identified using DNA microarray data from a rat liver fibrosis model. The expression level of each fibrosis associated marker gene was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) in clinical biopsy specimens from HCV-positive patients (n = 61). Analysis of changes in expression patterns and interactions of marker genes in functional categories was used to assess the biological mechanism of fibrosis. RESULTS: The profile data showed several biological changes associated with progression of hepatic fibrosis. Clustered genes in functional categories showed sequential changes in expression. Several sets of clustered genes, including those related to the extracellular matrix (ECM), inflammation, lipid metabolism, steroid metabolism, and some transcription factors important for hepatic biology showed expression changes in the immediate early phase (F1/F2) of fibrosis. Genes associated with aromatic amino acid (AA) metabolism, sulfur-containing AA metabolism and insulin/ Wnt signaling showed expression changes in the middle phase (F2/F3), and some genes related to glucose metabolism showed altered expression in the late phase of fibrosis (F3/F4). Therefore, molecular networks showing serial changes in gene expression are present in liver fibrosis progression in hepatitis C patients. CONCLUSION: Analysis of gene expression profiles from a perspective of functional categories or molecular networks provides an understanding of disease and suggests new diagnostic methods. Selected marker genes have potential utility for biological identification of advanced fibrosis.
基金supported by the Medical Science Foundation of Guangdong Health Department,No.B2011303the National Natural Science Foundation of China,No.30972843
文摘A rat model of diabetes mellitus was established by intraperitoneal injection of streptozotocin. Three days later, the rats were intraperitoneally administered 140 mg puerarin/kg daily, for a total of 60 successive days. DNA ladder results showed increased apoptosis over time in retinal pigment epithelial cells from rats with streptozotocin-induced diabetes mellitus. Western blot analysis, Reverse transcription-PCR, immunohistochemistry, and flow cytometry results showed increased expression of 3-nitrotyrosine, a peroxyntrite marker, as well as inducible nitric synthase and Fas/FasL, in retinal pigment epithelial cells. Puerarin reversed these changes, and results demonstrated that puerarin inhibited Fas/FasL expression and alleviated peroxyntrite injury to retinal pigment epithelial cells. These results suggested that puerarin inhibited production of inducible nitric oxide synthase and directly antagonized peroxyntrite injury in retinal pigment epithelial cells.
文摘BACKGROUND Anti-programmed death therapy has thrust immunotherapy into the spotlight.However,such therapy has a modest response in hepatocellular carcinoma(HCC).Epigenetic immunomodulation is a suggestive combinatorial therapy with immune checkpoint blockade.Non-coding ribonucleic acid(ncRNA)driven regulation is a major mechanism of epigenetic modulation.Given the wide range of ncRNAs that co-opt in programmed cell-death protein 1(PD-1)/programmed death ligand 1(PD-L1)regulation,and based on the literature,we hypothesized that miR-155-5p,miR-194-5p and long non-coding RNAs(lncRNAs)X-inactive specific transcript(XIST)and MALAT-1 are involved in a regulatory upstream pathway for PD-1/PD-L1.Recently,nutraceutical therapeutics in cancers have received increasing attention.Thus,it is interesting to study the impact of oleuropein on the respective study key players.AIM To explore potential upstream regulatory ncRNAs for the immune checkpoint PD-1/PD-L1.METHODS Bioinformatics tools including microrna.org and lnCeDB software were adopted to detect targeting of miR-155-5p,miR-194-5p and lncRNAs XIST and MALAT-1 to PD-L1 mRNA,respectively.In addition,Diana tool was used to predict targeting of both aforementioned miRNAs to lncRNAs XIST and MALAT-1.HCC and normal tissue samples were collected for scanning of PD-L1,XIST and MALAT-1 expression.To study the interaction among miR-155-5p,miR-194-5p,lncRNAs XIST and MALAT-1,as well as PD-L1 mRNA,a series of transfections of the Huh-7 cell line was carried out.RESULTS Bioinformatics software predicted that miR-155-5p and miR-194-5p can target PDL1,MALAT-1 and XIST.MALAT-1 and XIST were predicted to target PD-L1 mRNA.PD-L1 and XIST were significantly upregulated in 23 HCC biopsies compared to healthy controls;however,MALAT-1 was barely detected.MiR-194 induced expression elevated the expression of PD-L1,XIST and MALAT-1.However,overexpression of miR-155-5p induced the upregulation of PD-L1 and XIST,while it had a negative impact on MALAT-1 expression.Knockdown of XIST did have an impact on PD-L1 expression;however,following knockdown of the negative regulator of X-inactive specific transcript(TSIX),PD-L1 expression was elevated,and abolished MALAT-1 activity.Upon co-transfection of miR-194-5p with siMALAT-1,PD-L1 expression was elevated.Co-transfection of miR-194-5p with siXIST did not have an impact on PD-L1 expression.Upon co-transfection of miR-194 with siTSIX,PD-L1 expression was upregulated.Interestingly,the same PD-L1 expression pattern was observed following miR-155-5p cotransfections.Oleuropein treatment of Huh-7 cells reduced the expression profile of PD-L1,XIST,and miR-155-5p,upregulated the expression of miR-194-5p and had no significant impact on the MALAT-1 expression profile.CONCLUSION This study reported a novel finding revealing that opposing acting miRNAs in HCC,have the same impact on PD-1/PD-L1 immune checkpoint by sharing a common signaling pathway.
文摘AIM: To determine the gene expression profile data for the whole liver during development of dimethylni-trosamine (DMN)-induced hepatic fibrosis.METHODS: Marker genes were identified for different types of hepatic cells, including hepatic stellate cells (HSCs), Kupffer cells (including other inflammatory cells), and hepatocytes, using independent temporal DNA microarray data obtained from isolated hepatic cells. RESULTS: The cell-type analysis of gene expression gave several key results and led to formation of three hypotheses: (1) changes in the expression of HSC-specific marker genes during fibrosis were similar to gene expression data in in vitro cultured HSCs, suggesting a major role of the self-activating characteristics of HSCs in formation of fibrosis; (2) expression of mast cell-specific marker genes reached a peak during liver fibrosis, suggesting a possible role of mast cells in formation of fibrosis; and (3) abnormal expression of hepatocyte-specific marker genes was found across several metabolic pathways during fibrosis, including sulfur-containing amino acid metabolism, fatty acid metabolism, and drug metabolism, suggesting a mechanistic relationship between these abnormalities and symptoms of liver fibrosis. CONCLUSION: Analysis of marker genes for specific hepatic cell types can identify the key aspects of fibro-genesis. Sequential activation of inflammatory cells and the self-supporting properties of HSCs play an important role in development of fibrosis.
基金the Research Division of Qazvin University of Medical Sciences for supporting the project financiallythe Medicinal Plants Research Center,Institute of Medicinal Plants, (ACECR),Karaj,Iran
文摘Objective:To evaluate the scolicidal and immunomodulatory effect of the Ziziphora tenuior(Z.tenuior) extract and its fractions.Methods:Protoscolices were treated with six concentrations(3,5,10,25,50,and 100 mg/mL) of Z.tenuior extract and its fractions(ethanol,petroleum ether,ethyl acetate and chloroform) in periods of 10,20,30,40,50 and 60 minutes,and viability of protoscolices was evaluated using the 1.0%eosin.To examine the immunomodulatory effects of Ziziphora and its fractions on macrophage cells,the non-toxic concentration of extract and different fractions determined by MTT assay,and the Griess reaction was used to measure the level of nitrite as an indicator of nitric oxide by the macrophage cells in 10,100 and 200 μg/mL in 24 hours at 37 ℃.Results:In this study,the Z.tenuior extract at 10 mg/mL concentration was able to kill all protoscolices during 20 minutes.By increasing the concentration to 25 mg/mL,the scolicidal time reduced to 10 minutes.Regarding the effect of different fractions of Z.tenuior,the ethanolic fraction showed the highest scolicidal activity.The extract demonstrated an inhibitory effect on the activity of macrophages and reduced nitric oxide production.Although the petroleum ether and ethanolic fractions of the extract reduced nitric oxide production,nevertheless,this effect was only significant at 10 and 100 ug/mL concentrations(P<0.05).Conclusion:The Z.tenuior extract and its fractions were effective against protoscolices yet the effect of total extract was considerable.Our findings indicates that the extract and its ethanolic and petroleum ether fractions could have anti-inflammatory properties.
