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Objective To design and verify the reliability of a shoelace tensile test system. Methods Incremental loads of 0-196 N were applied to three tension sensors, each load was repeated nine times, with the load removed and interval of 30 s during the repeated tests. Then output voltage of the sensors under each load was collected. Linear regression analysis was used to explore linear relationship between the collected voltage signal and the incremental load. Accuracy, precision and consistency intervals were used to verify consistency of the measured values with the true load. Bland-Altman analysis and intra-group correlation coefficient (ICC) analysis were used to verify the repeatability and reliability of the tensile sensor. Results There was a significant linear correlation between output voltage signal of the sensors and the load (P< 0. 000 1, R2= 0. 999 9), and ICC of three sensors was above 0. 999 (P<0. 000 1). The mean values of the coefficients of variation of the measured values for three tensile sensors under different loads were 0. 003 8, 0. 002 2 and 0. 003 5, respectively. Conclusions The shoelace tensile test system has high reliability and can be used for real-time acquisition of shoelace tension.
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Objective:To evaluate the inhibitory effect of a retinoid derivative ECPIRM on proliferation of a cutaneous T-cell lymphoma (CTCL) cell line HH, and to explore its potential mechanisms.Methods:Cultured HH cells were treated with ECPIRM at different concentrations of 0 (control group) , 5, 10 and 20 μmol/L separately for 72 hours, cell counting kit-8 (CCK8) assay was performed to evaluate the effect of ECPIRM on the proliferative activity of HH cells, and flow cytometry to investigate the effect of ECPIRM on apoptosis of HH cells. Some HH cells were treated with 10 μmol/L ECPIRM for 72 hours, transcriptome sequencing was performed to investigate gene expression changes triggered by ECPIRM in HH cells, and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and gene ontology (GO) enrichment analysis were then performed to analyze differentially expressed genes in HH cells induced by ECPIRM. Reverse transcription-qPCR was subsequently conducted to verify changes in key gene expression in related pathways. Intergroup differences were analyzed by using one-way analysis of variance, and least significant difference (LSD) - t test was used for multiple comparisons. Results:CCK8 assay showed that the 50% inhibitory concentration (IC50) of ECPIRM on HH cells was 4.91 ± 2.48 μmol/L, the viability of HH cells significantly differed among the control group, and 5-, 10-and 20-μmol/L ECPIRM groups (100.00% ± 2.87%, 49.58% ± 4.53%, 48.36% ± 2.88%, 31.44% ± 2.46%, respectively, F=162.86, P < 0.001) , and was significantly lower in the 5-, 10-and 20-μmol/L ECPIRM groups than in the control group ( t=15.36, 15.73, 20.89, respectively, all P < 0.001) . Flow cytometry showed that there was a significant difference in the apoptosis rate among the 4 groups (11.51% ± 1.84%, 23.83% ± 5.72%, 36.19% ± 8.33%, 49.75% ± 4.10%, respectively, F=17.62, P < 0.001) , and the 10-and 20-μmol/L groups showed significantly increased apoptosis rates compared with the control group ( t=4.46, 6.92 respectively, both P < 0.01) . Transcriptomics analysis revealed that the inhibitory effect of ECPIRM on the cellular proliferative activity may be related to the metabolic regulation of steroids. As reverse transcription-qPCR revealed, the 10-μmol/L ECPIRM group showed significantly decreased mRNA expression of L-amino acid oxidase (IL4I1) , acetyl-coenzyme A acetyltransferase 2 (ACAT2) , 3-hydroxy-3-methylglutaryl-coenzyme A synthase 1 (HMGCS1) , mevalonate diphosphate decarboxylase (MVD) , 3-β-hydroxysteroid-8,7-isomerase (EBP) , very low-density lipoprotein receptor (VLDLR) , 3-hydroxy 3-methylglutaryl-CoA reductase (HMGCR) compared with the control group (all P < 0.05) . Conclusion:The retinoid derivative ECPIRM exerted marked anti-proliferative and apoptosis-inducing effects on HH cells, which may be related to the decreased expression of key genes involved in steroid metabolism.
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Efforts to control inflammation and achieve better tissue repair in the treatment of periodontitis have been ongoing for years. Human β-defensin 3, a broad-spectrum antimicrobial peptide has been proven to have a variety of biological functions in periodontitis; however, relatively few reports have addressed the effects of human periodontal ligament cells (hPDLCs) on osteogenic differentiation. In this study, we evaluated the osteogenic effects of hPDLCs with an adenoviral vector encoding human β-defensin 3 in an inflammatory microenvironment. Then human β-defensin 3 gene-modified rat periodontal ligament cells were transplanted into rats with experimental periodontitis to observe their effects on periodontal bone repair. We found that the human β-defensin 3 gene-modified hPDLCs presented with high levels of osteogenesis-related gene expression and calcium deposition. Furthermore, the p38 MAPK pathway was activated in this process. In vivo, human β-defensin 3 gene-transfected rat PDLCs promoted bone repair in SD rats with periodontitis, and the p38 mitogen-activated protein kinase (MAPK) pathway might also have been involved. These findings demonstrate that human β-defensin 3 accelerates osteogenesis and that human β-defensin 3 gene modification may offer a potential approach to promote bone repair in patients with periodontitis.
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Animales , Humanos , Ratas , Antiinfecciosos , Metabolismo , Farmacología , Diferenciación Celular , Células Cultivadas , Osteogénesis , Ligamento Periodontal , Metabolismo , Periodontitis , Quimioterapia , Ratas Sprague-Dawley , beta-Defensinas , Metabolismo , FarmacologíaRESUMEN
OBJECTIVE：To i mprove the transfer rate and purity of oleanolic acid and ursolic acid in total triterpenoids from Ligustrum lucidum ，so as to optimize the purification technology. METHODS ：Oleanolic acid and ursolic acid were used as representative components of total triterpenoids ，and their contents were determined by HPLC. The determination was performed on Thermo BDS Hypersil C 18 column with mobile phase consisted of methanol- 0.02％ ammonium acetate solution （80∶20，V/V）at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm，and column temperature was 30 ℃. The sample size was 20 μ L. In single factor tests，using transfer rate of oleanolic acid and ursolic acid as index ，the effects of water precipitation temperature and time ，the amount of redissolved ethanol on the purification technology was investigated ；using transfer rate and purity of two components as indexes ，the effects of the amount of activated carbon and volume fraction of crystallization ethanol were investigated. Based on it ，using the amount of redissolved ethanol and activated carbon ，volume fraction of crystallization ethanol as factors ，Box-Behnken response surface methodology was used to optimize the purification technology ，and validation tests were performed. RESULTS ：The optimal purification technology was adding 4-fold（mL/g，the same below ）water in L. lucidum concentrated solution ，placing for 2 hours at 0 ℃（water precipitation ）；adding 1-fold ethanol to dissolve （redissolution）； adding 4％ activated carbon （edulcoration）；finally adding water to adjust the volume fraction of ethanol to 80％，placing at 4 ℃ for 12 hours（crystallization），centrifuging and drying. The results of 3 times of validation tests showed that the transfer rates of oleanolic acid and ursolic acid in total triterpenoids prepared by optimized technology were 61.11％ and 65.78％，the purities of them were 53.44％ and 19.79％，and RSDs were both lower than 3％. CONCLUSIONS ：The optimized purification technology has high extraction efficiency and simple operation ，which can be used for industrial production of purification of total triterpenoids from L. lucidum and the development of corresponding preparations.
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Objective To investigate the clinical efficacy and safety of transcatheter arterial chemoemblization (TACE) using raltitrexed and lobaplatin in treating advanced hepatocellular carcinoma (HCC).Methods From March 2009 to November 2014,95 cases were treated by raltitrexed combined with lobaplatin (raltitrexed group) through TACE and 124 cases by fluorouracil combined with oxaliplatin (fluorouracil group) through TACE.Disease control rate (DCR),median progression-free survival (mPFS) time and median overall survival (mOS) time were compared between the two groups.Survival rate were analyzed using Kaplan-Meier method and Log-rank analysis in SPSS 16.0.Results The disease control rate of raltitrexed group was 91.6％ (87/95),compared with fluorouracil group of 84.6％ (105/124) in fluorouracil group (x2 =2.505,P =0.474).The mPFS of raltitrexed group was 6.8 months and that of fluorouracil group was 5.9 months (x2 =5.542,P =0.019);mOS of raltitrexed group was 13.6 months and fluorouracil group was 11.4 months (x2 =5.953,P =0.015).The main adverse reactions in the two groups were not statistically significant (P ＞ 0.05).Conclusions TACE using rahitrexed and oxaliplatin prolongs the progression free survival and overall survival time of patients with advanced hepatic carcinoma.
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Microtubules are the important components in cytoskeleton, which are found in all eukaryotic cells. Microtubules can assemble with the other proteins to form spindle, centriole and neural tube structure. Microtubules play important roles in cell mitosis, intracellular transportation, cell shape maintenance and signal transduction process. Microtubules are the important targets in many drug discoveries. The antitumor drugs acting on microtubules are the most effective clinical drugs, which can inhibit the polymerization of microtubules, destroy microtubules dynamic balance and spindle, block cell cycle and induce tumor cells necrosis. The paper re-viewed the effects and action mechanisms of microtubules targeted antitumor drugs, and discussed their application prospects in clinics.
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Objective To investigate the clinical efficacy and safety of transcatheter arterial chemoemblization (TACE) using raltitrexed and lobaplatin in treating advanced hepatocellular carcinoma (HCC).Methods From March 2009 to November 2014,95 cases were treated by raltitrexed combined with lobaplatin (raltitrexed group) through TACE and 124 cases by fluorouracil combined with oxaliplatin (fluorouracil group) through TACE.Disease control rate (DCR),median progression-free survival (mPFS) time and median overall survival (mOS) time were compared between the two groups.Survival rate were analyzed using Kaplan-Meier method and Log-rank analysis in SPSS 16.0.Results The disease control rate of raltitrexed group was 91.6％ (87/95),compared with fluorouracil group of 84.6％ (105/124) in fluorouracil group (x2 =2.505,P =0.474).The mPFS of raltitrexed group was 6.8 months and that of fluorouracil group was 5.9 months (x2 =5.542,P =0.019);mOS of raltitrexed group was 13.6 months and fluorouracil group was 11.4 months (x2 =5.953,P =0.015).The main adverse reactions in the two groups were not statistically significant (P ＞ 0.05).Conclusions TACE using rahitrexed and oxaliplatin prolongs the progression free survival and overall survival time of patients with advanced hepatic carcinoma.
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Objective To estimate the effect of a tretinoin derivative ECPIRM on retinoic acid receptors (RARs),and to observe skin irritation responses to it in mice.Methods Cultured SCL-1 cells were divided into 2 groups to be treated with culture medium containing 10 μmol/L ECPIRM (ECPIRM group) or 10 μmol/L all-trans retinoic acid (ATRA) (ATRA group) for 24 hours,and those treated with drug-free culture medium served as the control group.Western blot analysis and real-time fluorescence-based quantitative PCR were performed to quantify the protein and mRNA expressions of RARs (RARα,RARβ,RARγ and RXRα) respectively.In addition,real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expressions of two target genes of the activated RAR signaling pathway,i.e.,cytochrome P450 26A1 (CYP26A1) and tazarotene-induced gene 1 (TIG1).Eight BALB/c mice were equally divided into 2 groups to be topically treated with 0.075％ ECPIRM gel or 0.05％ ATRA cream at equal molar concentrations on the shaved skin once daily for 21 successive days.Skin irritation reactions were assessed in these mice.Results Compared with the control group,the ATRA group showed significantly increased protein and mRNA expressions of RARα,RARβ and RARγ (all P ＜ 0.05).The mRNA expressions of CYP26A1 and TIG1 genes in the ATRA group were 25.49 and 3.88 times that in the control group respectively (both P ＜ 0.01).However,there was no significant difference in the protein expressions of RARα,RARβ,RARγ and RXRα,or mRNA expressions of RARα,RARβ,RARγ CYP26A1 and TIG1 between the ECPIRM group and control group (all P ＞ 0.05).Obvious Skin irritation reactions such as erythema and desquamation were observed in BALB/c mice after 2-day topical treatment with ATRA cream,and their degree peaked after 5-day treatment.However,neither erythema nor desquamation was observed in BALB/c mice during 21-day treatment with 0.075％ ECPIRM gel.Conclusion Unlike ATRA,ECPIRM cannot activate the canonical RAR signaling pathway or cause skin irritation reactions.
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Objective To investigate the effect of adenovirus-mediated IL-24 (Ad-IL-24)gene expression on the apoptosis in a human squamous cell carcinoma cell line COLO 16, and to explore the underlying molecular mechanism. Methods Cultured COLO 16 cells were divided into two groups to be transfected with an adenovirus vector carrying the IL-24 gene (Ad-IL-24 group)or green fluorescent protein (Ad-GFP group), while those receiving no treatment served as the control group. After culture for different durations, qPCR was performed to quantify IL-24 gene expression, methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferative activity of COLO 16 cells, flow cytometry to detect the apoptosis of COLO 16 cells, laser scanning confocal microscopy (LSCM) to observe the morphological changes of COLO 16 cells, Western blot to determine the levels of Bax and Bcl-2 proteins and to evaluate the activation of caspase-3, qPCR to determine the levels of Bax and Bcl-2 mRNAs, an immunofluorescence assay to observe the expression of Bax and Bcl-2 proteins. Statistical analysis was carried out by a two-sample t-test with the SPSS 19.0 software. Results MTT assay showed that the proliferation of COLO 16 cells in the Ad-IL-24 group was significantly inhibited as early as 4 days after the transfection; thereafter, the inhibitory effect increased in a time-dependent manner, and peaked on day 6(P0.05). Flow cytometry revealed that the apoptosis rate was significantly higher in the Ad-IL-24 group(13.10%± 0.92%)than in the control group(3.69%± 0.36%, P0.05). LSCM demonstrated that the apoptosis of COLO 16 cells was accelerated in the Ad-IL-24 group. The immunofluorescence assay, Western blot and qPCR all showed that the mRNA and protein expressions of Bax were increased, but those of Bcl-2 were decreased in the Ad-IL-24 group compared with the Ad-GFP group and control group. Moreover, Western blot showed a protein band that could specifically bind to the anti-cleaved caspase-3 antibody in the Ad-IL-24 group, but not in the Ad-GFP group or control group. Conclusions Ad-IL-24 can induce apoptosis in human COLO 16 squamous cell carcinoma cells, probably by up-regulating Bax expression, down-regulating Bcl-2 expression, and activating caspase 3.
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Objective To evaluate the efficacy and safety of uterine artery chemoembolization(UACE)in conjunction with dilata-tion and curettage in the treatment of cervical pregnancy.Methods UACE was performed in 7 patients with cervical pregnancy,after bilateral uterine artery perfusion of methotrexate,using gelatin sponge for embolization.Dilatation and curettage was carried out within 1 week after the procedure.Results The symptom of vaginal bleeding was effectively controlled after UACE,then dilatation and curettage performed in 6 within 1 week,the blood loss during curettage procedure was 10-30 mL(mean 21 mL).The serumβ-HCG decreased to normal range within 1 week in 1 and 3 weeks in 6 after intervention,average 1 6.4 ± 6.9 days.No serious com-plications occurred and follow up did not show symptoms of premature ovarian failure.Conclusion UACE combined with dilatation and curettage significantly improve the efficacy and safety of the treatment of cervical pregnancy,and avoid serious incident,such as massive hemorrhage and unnecessary resection,the clinical application is safe and effective.
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<p><b>OBJECTIVE</b>To establish a method for quantitative analysis of gastrodin, baicalin, paeonolum and pinoresinol diglucoside in Tiangou capsules by HPLC.</p><p><b>METHOD</b>The separation was performed on SinoChrom ODS-BP column (4.6 mm x 150 mm, 5 microm). The mobile phase consisted of acetonitrile and 0.2% phosphoric acid aqueous solution with gradient elution program. The flow rate was 1.0 ml x min(-1) and the column temperature was 30 degrees C. UV detection wavelength was set at 230 nm.</p><p><b>RESULT</b>The linear ranges of gastrodin, baicalin, paeonolum and pinoresinol diglucoside were 0.1168-1.1680, 0.1142-1.1420, 0.0512-0.5120, 0.0556-0.5560 microg, respectively. The average recoveries of gastrodin, baicalin, paeonolum and pinoresinol diglucoside were 100.3% (RSD 0.53%), 99.96% (RSD 1.15%), 99.90% (RSD 1.17%), 99.97% (RSD 1.62%), respectively.</p><p><b>CONCLUSION</b>The method is accurate, simple, feasible and reliable, and is available for the quality control of Tiangou capsules.</p>
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Alcoholes Bencílicos , Química , Cápsulas , Cromatografía Líquida de Alta Presión , Métodos , Medicamentos Herbarios Chinos , Química , Flavonoides , Química , Furanos , Química , Glucósidos , Química , Lignanos , Química , Modelos Lineales , Compuestos Orgánicos , Reproducibilidad de los Resultados , Factores de TiempoRESUMEN
Objective To analyze the complete genome sequence of Guangxi HEV isolate swGX32 and to compare it with other HEV isolates. Methods The overlapping fragments of HEV isolate swGX32 were amplified with reverse-transcription nested polymerase chain reaction (RT-nPCR),and the 5′ and 3′ ends of viral genome were amplified with rapid amplification of cDNA ends (RACE). The PCR products were cloned and sequenced. The sequence and phylogenetic analysis of swGX32 was performed. Results The genome of swGX32 consisted of 7240 nt excluding the polyA tail, with 4 nt overlapping between ORF1 and ORF2. ORF3 is contained in the sequence of ORF2. The complete genome sequence of swGX32 shared identity of 73%-74%, 73%, 74%-75%,83%-94% with HEV genotype 1,2,3 and 4, respectively. Among all these HEV reference sequences, swGX32 showed the highest identity with the human isolate JKO-ChiSai98C (94%). Phylogenetic tree showed that swGX32 belonged to genotype 4 and clustered with JKO-ChiSai98C in the branch of HEV subtype 4a. Conclusion The swine HEV isolate swGX32 is closely related to human strain JKO-ChiSai98C genetically and phylogenetically, which further provides molecular biology evidence of hepatitis E as a zoonosis.
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Objective To study the effects of paeonol on the tyrosinase activity and melanogenesis in co-culture model of human melanocytes and keratinocytes. Methods Melanocytes and the co-culture model of human melanocytes and keratinocytes were cultivated and the proliferation of melanocytes and the co-cultures was measured by MTT colorimetric assay. The tyrosinase activity and melanin level were measured by enzymic method. Results The melanin synthesis and tyrosinase activity were markedly suppressed by paeonol in a dose-dependent manner at the concentrations of 50?mol/L, 100?mol/L, and 200?mol/L in both melanocytes and co-cultures. The significant stronger suppression was observed with 100?mol/L and 200?mol/L of paeonol than that with controls (P
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Objective To evaluate the inhibitory action of ethanolic extracts from196kinds of Chi-nese herbs on tyrosinase.Methods Tyrosinase inhibitory activity was determined by the dopachrome method using L-DOPA as the substrate and the amount of dopachrome in the reaction mixture was measured by spec-trophotometer.Results Nine Chinese crude extracts[Glycyrrhiza glabra L.,Melaphis chinensis(Bell.)Baker,Cryptotympana atrata Fabricius,Paeonia suffruticosa Andr.,Sophora flavescens Ait.,Spirea thunbegii Sieb.et Bl.,Xanthium sibiricum Patr.,Morus alba L.,Rheum palmatum L.]showed potent inhibitory action on tyrosi-nase compared with positive control arbutin(1mmol/L,P0.05).Conclusion The results suggest that20kinds of crude extracts from Chinese herbs[Glycyrrhiza glabra L.,Melaphis chinensis(Bell.)Baker,Cryptotympana a-trata Fabricius,Paeonia suffruticosa Andr.,Sophora flavescens Ait.,Spirea thunbegii Sieb.et Bl.,Xanthium sibiricum Patr.,Morus alba L.,Rheum palmatum L.,Artemisia anomala S.Moore,Hemistepta lyrata Bunge,Lycium chinensis Mill.,Dipsacus asper wall.,Gastrodia elata Bl.,Forsythia suspensa(Thunb.)Vahl.,Acer gin-nala Maxim.,Glycyrrhiza uralensis Fisch,Patrinia sabiosaefolia Fisch.,Buxus sinica(Rehd.et Wils.)Ching,Lonicera japonica Thunb.,]inhibit tyrosinase and may be used as depigmentaty agents for pigmentary skin dis-eases caused by abnormal tyrosinase activity.
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AIM: To investigate the method for using semi-bionic extraction (SBE) on Simiao Yongan Decoction as an example. METHODS: Orthogonal design was used to optimize the extraction conditions of SBE with the evaluation markers such as chlorogenic acid, ferulic acid, glycyrrhizic acid, total polysaccharide, ethanol soluble extractives and dried extract. RESULTS: The optimized SBE conditions were as follows: The medical material was powdered through a sieve with 4 meshes) and extracted with 20% ethanol for three times, the pH of the three extractions were in proper order 3.50, 7.50, 8.50 ; the solvent volume was 10,8,8 times, respectively, and the extraction time was 1.5,1.0,1.0 h, respectively. CONCLUSION: The optimum extraction conditions are stable and practicable, and surpass the original SBE extraction conditions.