Determination of 1-methoxy-2-propanol in urine by headspace solid-phase microextraction coupled with gas chromatography / 中华劳动卫生职业病杂志

Objective@#To establish a method for the determination of 1-methoxy-2-propanol in urine using headspace solid phase micro-extraction coupled with gas chromatography.@*Methods@#The 1-methoxy-2-propanol was enriched by headspace solid phase micro-extraction fiber coated with carbene/polydimethylsiloxane (CAR/PDMS) . Single factor rotation method was used to optimize the conditions of extraction temperature, salt amount, and extraction time. The separation was performed on DB-5 (30 m×0.32 mm×0.25 μm) capillary column and detected with flame ionization detector. The quantification was based on the standard curve.@*Results@#The concentration of 1-methoxy-2-propanol in urine was linear in the range of 0.50-10.0 mg/L, and the linear correlation coefficient was 0.9993. The detection limit of the method was 0.14 mg/L, and the limit of quantification was 0.45 mg/L. The recovery was 85.8% to 104.7%, and the RSD of intra- and inter-batch precision were 3.25%-6.65% and 0.81%-3.96%, respectively.@*Conclusion@#The method is high sensitivity and simple operation, and is suitable for the determination of 1-methoxy-2-propanol in urine of occupational exposure population.

One new phenylpropanoid from Hainan Ceriops tagal / 中草药

Objective: To study the chemical constituents from the ethyl acetate fraction of Ceriops tagal. Methods: Compounds were isolated and purified by various column chromatography, recrystallization, and HPLC. And their structures were elucidated by physicochemical property and spectral analysis. Results: Eight compounds were isolated from the ethyl acetate fraction of Ceriops tagal and identified as (E)-methyl-3-(5-hydroxy-2,3,4-trimethoxyphenyl)acrylate (1), tagalphenylpropinoidin A (2), coniferyl aldehyde (3), 2,3-dimethoxy-5-(1-propenyl)phenol (4), methyl syringate (5), syringaldehyde (6), pinosylvin monomethyl ether (7), and n-hexadecane acid (8), respectively. Conclusion: Compound 1 is a new phenylpropanoid, named as tagalphenylpropanoidin C, and compounds 4-8 are obtained from C. tagal for the first time.

Rheum officinale at different years old stage / 中草药

Objective To analyze the content and variation rules of 10 constituents in radix, rhizome, and leaf of Rheum officinale at one-, two-, and three-year-old stage, respectively, and provide theoretical guidance for efficient production and quality control of the crud drug. Methods The content of each constituent in R. officinale was determined by high performance liquid chromatography (HPLC), and one factor analysis of variance and multiple comparison were performed by SPSS 24.0. Results HPLC system was established for the determination of 10 components in R. officinale. The linear range was good (r2 > 0.997), RSD of precision, stability, and repeatability were less than 2%, and the recoveries were 96.10%—107.10%, respectively. The content analyses showed that, in the same part, the content of gallic acid decreased significantly year by year or at the 2nd growth years (P 3 > 2 (P 1 > 2 (P radix > leaf (P < 0.05). Conclusion The HPLC based determination of 10 constituents in R. officinale showed that the accumulation profiles of the samples at different years or from different parts varied. For the same parts, the contents of most constituents increased year by year. During the same growth year, the contents of most constituents in radix or rhizome were higher than those in leaf. The radix and rhizome of the three years old samples had the highest contents of main constituents.

Preparation and Characterization of Celecoxib-loaded Micelles / 中国药师

Objective: To prepare celecoxib-loaded micelles with polyethylene glycol monomethyl ether-polylactic acid ( mPEG-PLA) block copolymer as the carrier material and evaluate the physical and chemical properties .Methods:Celecoxib-loaded micelles were prepared by a film dispersion method .The micelle formula and preparation process were screened by single factor experiment and further optimized by Box-Behnken response surface method .The physical and chemical properties of celecoxib-loaded micelles such as microscopic morphology , particle size distribution and zeta potential were evaluated .The in vitro drug release of celecoxib-loaded mi-celles was investigated by dynamic membrane dialysis .Results:Celecoxib-loaded micelles prepared according to the optimized formula showed the following properties:the particle size distribution was (35.6 ±15.1) nm, PdI was (0.152 ±0.05), and the zeta potential was (-24.6 ±2.9) mV.In 0.5％SDS phosphate buffered saline (pH 6.8), the in vitro cumulative release of celecoxib-loaded mi-celles reached up to 81 .5％in 24 h.Conclusion:It is simple and feasible to prepare celecoxib-loaded micelles by the thin film dis-persion method .

Effect and mechanism of photodynamic therapy on dental caries in rats / 新乡医学院学报

Objective To evaluate the effect of photodynamic therapy (PDT) on dental caries in rats,and to investigate the mechanism.Methods Forty male Wistar rats (21 days old,specific pathogen-free,weaned) with dental caries induced by Streptococcus mutans were randomly divided into saline group,hematoporphyrin monomethyl ether (HMME)-PDT group,simple laser group,simple photosensitizer group and sodium fluoride group,with eight rats in each group.The molars of rats in the saline group were smear repeatedly for 5 minutes with cotton balls containing 150 μL normal saline.The molars of rats in HMME-PDT group were incubated for 5 minutes without light by placing the cotton balls containing 150 μL HMME (40 mg · L-1) on the tooth surface,then molars were irradiated vertically by 532 nm semiconductor laser,the power density was 140 mW · cm-2,the irradiation time was 90 s,and the energy density was 12.6 J · cm-2.The molars of rats in the simple laser group were irradiated vertically by 532 nm semiconductor laser;the irradiation time,power density and energy density were the same as those in HMME-PDT group;but the molars were not treated with HMME.The molars of rats in the simple photosensitizer group were smear repeatedly for 5 minutes with cotton balls containing 150 μL HMME (40 mg · L-1),but the molars were not irradiated by laser.The molars of rats in the sodium fluoride group were smear repeatedly for 5 minutes with cotton balls containing 150 μL fluorinse (2 g · L-1).The rats in each group were treated once a week for 4 weeks.Streptococcus mutans were collected from oral cavity of rats after each treatment,and bacterial inhibition was recorded by plate count.The rats were killed after 4 weeks,and the Keyes caries score was recorded.The surface morphology of the teeth was observed by scanning electron microscope.Results The time factor had no effect on the growth of oral Streptococcus mutans (F =0.11,P ＞ 0.05),the treatment factor had effect on the growth of oral Streptococcus mutans (F =230.89,P ＜ 0.05),and there was interaction effect between treatment factor and time factor (F =6.36,P ＜ 0.05).The colony number of Streptococcus mutans in HMME-PDT group and sodium fluoride group was lower than that in saline group,photosensitizer group and simple laser group at the first,second,third and fourth week after treatment (P ＜ 0.05).The colony number of Streptococcus mutans in HMME-PDT group was lower than that in sodium fluoride group at the first,second and third week after treatment (P ＜ 0.05).The Dx caries lesion of smooth surface caries wasn't found in each group.The Dm caries lesion of smooth surface caries wasn't found in the HMME-PDT group and sodium fluoride group.The Keyes scores of E and Ds caries lesion of smooth surface caries in the HMME-PDT group and sodium fluoride group was lower than that in the saline group,simple photosensitizer group and simple laser group(P ＜0.05),but there was no significant difference in the Keyes scores of E and Ds caries lesion of smooth surface caries between the HMME-PDT group and sodium fluoride group(P ＞0.05).The Dx caries lesion of pit and fissure caries was found in the saline group only.The Keyes score of E,Ds and Dm caries lesion of pit and fissure caries in the HMME-PDT group and sodium fluoride group was lower than that in the saline group,simple photosensitizer group and simple laser group (P ＜ 0.05),but there was no significant difference in the Keyes scores of E,Ds and Dm caries lesion of pit and fissure caries between the HMME-PDT group and sodium fluoride group(P ＞ 0.05).The results of scanning electron microscopy showed that the enamel surface was rough,uneven and there was a large number of scratch in the saline group and simple photosensitizer group.Relatively the enamel surface in the simple laser group was smooth,and there was a small circular scallops and dents was found in the structure;in the sodium fluoride group,the enamel surface was relatively smooth,take off the mining area was less,and remineralization was found.In the HMME-PDT group,the enamel surface was smooth,pitting was reduced and take off the mining area was small.Conclusion HMME-PDT had significant inhibitive efficacy on caries development in rats by reducing the number of cariogenic bacteria and improving the structure of the teeth.

Construction and evaluation of charge-reversal nanocarrier loaded with hematoporphyrin monomethyl ether / 药学学报

Charge-reversal nanocarrier was constructed to enhance lysosomal escape and improve an-titumor effect. We synthesized the cholesterol-polyethyleneimine-hexahydrophthalic anhydride (Chol-PEI-HHPA) polymer and characterized by 1H NMR. The charge-reversal liposomes (Lipo-HHPA) were synthesized and the hematoporphyrin monomethyl ether (HMME) was loaded. pH-triggered charge conversion was determined at different pH values. The lysosomal escape and cytotoxicity of the Lipo-HHPA were evaluated in MCF-7 cells. The Lipo-HHPA was uniform with an average particle size of 102 nm. Upon the irradiation of ultrasound, burst release of HMME could be observed. The zeta potential of Lipo-HHPA changed sharply from negative (-23.5 mV) to positive (+21.2 mV) over the pH range of 7.4-4.5. In the cellular uptake experiment, the lysosomal escape of Lipo-HHPA was observed. HMME loaded Lipo-HHPA displayed obviously enhanced cytotoxicity towards MCF-7 cells. These results indicate that the charge-reversal liposomes hold a great potential in improving the cytotoxicity and antitumor effect.

Hematoporphyrin monomethyl ether combined with He-Ne laser irradiation-induced apoptosis in canine breast cancer cells through the mitochondrial pathway

Hematoporphyrin monomethyl ether (HMME) combined with He-Ne laser irradiation is a novel and promising photodynamic therapy (PDT)-induced apoptosis that can be applied in vitro on canine breast cancer cells. However, the exact pathway responsible for HMME-PDT in canine breast cancer cells remains unknown. CHMm cells morphology and apoptosis were analyzed using optical microscope, terminal deoxynucleotidyl transferase dUTP nick end labeling fluorescein staining and DNA ladder assays. Apoptotic pathway was further confirmed by Real-time-polymerase chain reaction and Western blotting assays. Our results showed that HMME-PDT induced significant changes in cell morphology, such as formation of cytoplasmic vacuoles and the gradual rounding of cells coupled with decreased size and detachment. DNA fragmentation and cell death was shown to occur in a time-dependent manner. Furthermore, HMME-PDT increased the activities of caspase-9 and caspase-3, and released cytochrome c from mitochondria into the cytoplasm. HMME-PDT also significantly increased both mRNA and protein levels of Bax and decreased P53 gene expression in a time-dependent manner, while the mRNA and protein expression of Bcl-2 were repressed. These alterations suggest that HMME-PDT induced CHMm cell apoptosis via the mitochondrial apoptosis pathway and had anti-canine breast cancer effects in vitro.

Dose effect of laser irradiation in photodynamic therapy for laryngocarcinoma in BALB/c mouse / 国际生物医学工程杂志

Objective The dose and type of light and photosensitizer could seriously affect the curative effect of photodynamic therapy (PDT).The purpose of this study was to observe whether or not PDT with hematoporphyrin monomethyl ether (HMME) can cure laryngocarcinoma in the solid tumor model,and to define the proper laser amount for killing the cancer cells.Methods Forty eight BALB/c mouse models with subcutaneous Hep-2 laryngeal carcinomas were prepared.Mice were divided into six groups depending on the amount of laser received from 30 J/cm2 to 480 J/cm2 including a control group,tumor size in each group was between 8 mm and 10 mm.Tail vein injection were given with HMME prior to applying the laser light,and then illumination was carried out on the tumor at 3 h after HMME administration.Tumor volume,animal weight and histopathologic changes were observed after PDT.Results All mice apparently showed positive results via PDT,and the cancer had been cured in 120 J/cm2 and 480 J/cm2 groups.The laryngeal cancer lesions began to form scab 1 d after PDT and the scab became hard and black after 5 d.The tumor regression began simultaneously and completed around 30 d after PDT.Conclusions PDT may treat laryngeal cancers which sized less than 10 mm in mouse models.The optimum energy to destruct the laryngeal cancer cells may be 120 J/cm2.

Nature monomethyl ether derivatives of inositol:research progress / 国际药学研究杂志

Inositol,a hexol widely found in nature with a wide range of physiological and biochemical effects,can promote fat metabolism in liver and other tissues and lower blood cholesterol and thus has been used for the clinical treatment of fatty liver, cirrhosis,atherosclerosis and hyperlipidemia. Some derivatives of inositol have been reported to have different biological activities including hypoglycemic,antitussive,anti-inflammatory and other effects. In recent years,its monomethyl ether derivatives,such as pinitol,quebrachitol,and sequoyiol,have been studied widely. In this paper,the bioactivity,extraction,preparation and applications of these monomethyl ether derivatives of inositol are reviewed.

Impact of HMME-PDT on Pg of the surface of implants and cementum tablets / 国际生物医学工程杂志

Objective To evaluate the bactericidal effect of diode laser on Porphyromonas gingivalis (Pg),and to explore an optimized protocol for a safe dose of photodynamic therapy (PDT) to eliminate periodontal pathogens as well as the impact on the implant surfaces,so as to provide theoretical and experimental basis for PDT in periimplantitis therapy.Methods Artificial in vitro models were formatted by culturing Pg standard strain and ITI (International Team for Implantology) implants together in CDC broth.Then artificial in vitro models were treated by different doses of hematoporphyrin monomethyl ether (HMME) and different energy density of laser (EDL) for 60 s.The cultures were counted by colony form unit (CFU),and SPSS 17.0 statistical software was used for data statistical analysis to select the best EDL and HMME dose.Finally,ITI implants were observed by scanning electron microscope (SEM) to evaluate the impact of HMME-PDT on Pg of implant surfaces.Results When EDL was 12 J/cm2 and mass concentration of HMME was 25 μg/ml,SEM observations showed that PDT could effectively kill Pg ((13.00±5.00) CFU)without damaging the implant surfaces.Conclusions PDT therapy combining 630 nm diode laser with photosensitizer HMME have good bactericidal effect on Pg,and the EDL and HMME dose is as small as the clinical applicable safe dose.

Nature monomethyl ether derivatives of inositol: Research progress / 国际药学研究杂志

Inositol, a hexol widely found in nature with a wide range of physiological and biochemical effects, can promote fat metabolism in liver and other tissues and lower blood cholesterol and thus has been used for the clinical treatment of fatty liver, cirrhosis, atherosclerosis and hyperlipidemia. Some derivatives of inositol have been reported to have different biological activities including hypoglycemic, antitussive, anti-inflammatory and other effects. In recent years, its monomethyl ether derivatives, such as pinitol, quebrachitol, and sequoyiol, have been studied widely. In this paper, the bioactivity, extraction, preparation and applications of these monomethyl ether derivatives of inositol are reviewed.

Toxigenic profile and AFLP variability of Alternaria alternata and Alternaria infectoria occurring on wheat

The objectives of this study were to evaluate the ability to produce alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TA) by A. alternata and A. infectoria strains recovered from wheat kernels obtained from one of the main production area in Argentina; to confirm using AFLPs molecular markers the identify of the isolates up to species level, and to evaluate the intra and inter-specific genetic diversity of these two Alternaria species. Among all the Alternaria strains tested (254), 84% of them were able to produce mycotoxins. The most frequent profile of toxin production found was the co-production of AOH and AME in both species tested. TA was only produced by strains of A. alternata. Amplified fragment polymorphism (AFLPs) analysis was applied to a set of 89 isolates of Alternaria spp (40 were A. infectoria and 49 were A. alternata) in order to confirm the morphological identification. The results showed that AFLPs are powerful diagnostic tool for differentiating between A. alternata and A. infectoria. Indeed, in the current study the outgroup strains, A. tenuissima was consistently classified. Characteristic polymorphic bands separated these two species regardless of the primer combination used. Related to intraspecific variability, A. alternata and A. infectoria isolates evaluated seemed to form and homogeneous group with a high degree of similarity among the isolates within each species. However, there was more scoreable polymorphism within A. alternata than within A. infectoria isolates. There was a concordance between morphological identification and separation up to species level using molecular markers. Clear polymorphism both within and between species showed that AFLP can be used to asses genetic variation in A. alternata and A. infectoria. The most important finding of the present study was the report on AOH and AME production by A. infectoria strains isolated from wheat kernels in Argentina on a semisynthetic media for the first time. Also, specific bands for A. alternata and A. infectoria have been identified; these may be useful for the design of specific PCR primers in order to differentiate these species and to detect them in cereals.

Preparation and drug-loading property of phosphatidyl polyethyleneglycol monomethyl ether liposome / 国际生物医学工程杂志

Objective To develop liposome loaded with paclitaxel for the treatment of lung cancer,novel phosphatidyl polyethyleneglycol monomethyl ether liposomes were prepared and studied.Methods A series of phosphatidyl polyethyleneglycol monomethyl ether liposomes were synthesized.Liposomes with or without drug were prepared by ultrasound.The entrapment efficiency and size of drug loaded liposomes were evaluated.The influence of liposomes to A549 lung cancer cell was investigated by MTT method.The cell uptake of liposome was observed by laser confocal scan microscope.Results The liposome loaded with paclitaxel had an entrapment efficiency of 83％ and size of 100～200 nm.The MTT results showed phosphatidyl polyethyleneglycol monomethyl ether liposomes loaded paclitaxel have inhibition effect to A549 cells with long circulation,while liposomes without paclitaxel have not inhibition effect.Conclusion Drug loaded polyethyleneglycol monomethyl ether liposomes have sustained delivery to paclitaxel.They have potential application in the therapy of cancer.

Investigating the killing effect of photodynamic therapy induced by different photosensitizers on human leukemia cell in vitro / 国际生物医学工程杂志

Objective To investigate and compare the killing effect of photodynamic therapy (PDT)induced by hematoporphyrin derivative (HpD),hematoporphyrin monomethyl ether (HMME) and photocarcinorin (PsD007) on human leukemia cells K562 in vitro.Methods Human leukemia cells were cultured with serial concentrations of photosensitizers followed by irradiation of different dosage of laser light,then MTT colorimetric assay was applied to measure the relative survival rate of PDT for the cells.Results Significant difference in the inhibitory between the PDT group and control group was observed (P＜0.05).The survival rate of PDT for the cells elevated along with the increase in the concentration of sensitizer and dose of laser light.When the photosensitizer concentration was bigger (25 μg/ml) or the energy density was bigger (7.2 J/cm2),the effect of PsD007 was better than HMME,and they were significantly better than HpD (P＜0.05).Conclusion PDT has significant killing effect on human leukemia cells K562,and its relative inhibitory rate appears to be correlated with the dose of sensitizer and laser light irritation.The effect of PDT is related to the photosensitizers.The effect of HpD-PDT is not as effective as PsD007 and HMME.On the conditions of higher energy density and larger photosensitizer concentration,the effect of PsD007-PDT is better than HMME-PDT.

The inhibitory effect of photodynamic therapy and of an anti-VCAM-1 monoclonal antibody on the in vivo growth of C6 glioma xenografts

We investigated the effect of photodynamic therapy (PDT) and of an anti-vascular cell adhesion molecule-1 (VCAM-1) monoclonal antibody on the in vivo growth of C6 glioma. Seven days after inoculation with C6 cells, adult male Wistar rats weighing 280-300 g with MRI-confirmed glioma were randomly assigned to 4 groups (N = 15 per group): PDT + VCAM-1 antibody group; PDT group; VCAM-1 antibody group; control group. Eight days after inoculation, hematoporphyrin monomethyl ether (HMME) was administered as a photosensitizer and PDT was performed at 630 nm (illumination intensity: 360 J/cm²) for 10 min. VCAM-1 antibody (50 µg/mL) was then administered (0.5 mL) through the tail vein every other day from day 8 to day 16. At day 21, 5 rats in each group were sacrificed and cancers were harvested for immunohistochemistry and Western blot assay for the detection of VCAM-1, and TUNEL assay was used to detect apoptosis. Survival and tumor volume were recorded in the remaining 10 rats in each group. In the PDT group, tumor growth was significantly suppressed (67.2 percent) and survival prolonged (89.3 percent), accompanied by an increase in apoptosis (369.5 percent), when compared to control. Furthermore, these changes were more pronounced in the PDT + VCAM-1 antibody group. After PDT, VCAM-1 expression was markedly increased (121.8 percent) and after VCAM-1 monoclonal antibody treatment, VCAM-1 expression was significantly reduced (58.2 percent). PDT in combination with VCAM-1 antibody can significantly inhibit the growth of C6 glioma and prolong survival. This approach may represent a promising strategy in the treatment of glioma.

Apoptosis of hypertrophic scar fibroblasts induced by HMME-PDT / 中华医学美学美容杂志

Objective To investigate the apoptotic effects of hypertrophic scar fibroblast (HSF) induced by HMME-PDT.Methods Fibroblasts were cultured from nontreated hypertrophic scars,and cells at passages 4-6 were used for the experiments (photosensitizer dose 4 μg/ml,λ630 nm,pow er density 10 mw/cm2,energy fluence 2.5 J/cm2).Morphological and biochemical changes in fibroblasts were assessed by Hoechst 33258 staining and fluorescence microscopy.The rate of apoptotic or necrotic cells was detected by flow cytometry (FCM) through double staining of Annexin V -FITC and popodium iodide (PI),respectively.Results Marked morphological features of cell apoptosis were viewed under the fluorescent microscope through Hoechst 33258 staining.The analysis of FCM indica ted that the apoptotic rate was significantly increased after HMME PDT [(34.82 ± I.42) ％ vs (3.12±0.28) ％,P＜0.05],and apoptotic rate was higher than necrosis rate [(14.65±1.02) ％ vs (34.82±1.42) ％,P＜0.05].Conclusions Low level exposure to 630 nm PDT mediated by HMME appears to induce fibroblast apoptosis.

Inhibitive effect of photodynamic therapy with different parameters on caries development in rats / 国际生物医学工程杂志

ObjectiveTo evaluate the inhibitive effect of Photodynamic therapy (PDT) with different parameters on dental caries formation in rats.MethodsWistar rats were infected with S.mutans to establish the rat caries mode 1.The rat's teeth were treated with 0.9% NS and 0.2% NaF as controls;laser,photosensitizer and laser plus photosensitizer,respectively as treated groups.Hematoporyrin monomethyl ether(HM ME) was used as photosensitizer and laser as the light source.The rats were sacrificed after five weeks.Keyes count was used to measure the inhibitory effect of PDT on dental caries.The surface morphology of the teeth was evaluated by scan electronic microscopy(SEM).Results The PDT-treated group and NaF-treated group showed significantly reduced Keyes scores compared with the negative controls (P＜0.05).The laser-treated group,photosensitizer-treated group and NS-treated group showed no significantly reduced Keyes scores compared with the negative controls (P＜0.05).The differences of Keyes scores between various parameter PDT treated groups were significant (P＜0.05).Additionally,Keyes scores reduced gradually with the increasing of laser power.The outcome of SEM showed that the teeth treated with different laser powers exhibited different changes in morphology.Conclusion HMMEPDT had significant inhibitive efficacy on caries development in rats and its inhibitory effect presented close correlation with the power of laser,which indicates that it is important to apply appropriate power in clinical treatment.

Distribution of Hematoporphyrin Monomethyl Ether in Tumor-bearing Mice / 第二军医大学学报

spleen(?)heart(?)muscle.The pharmacokinetics of PsD-044 in mice was fitted by an open three compartment model. The pharmacokinetic parameters calculated were shown to be essentially consistent with the determination of tissue distribution. It was also shown that PsD-044 can't pass through blood-brain barrier.