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Chemical investigation of the culture extract of an endophytic Penicillium citrinum from Dendrobium officinale, afforded nine citrinin derivatives (1-9) and one peptide-polyketide hybrid GKK1032B (10). The structures of these compounds were determined by spectroscopic methods. The absolute configurations of 1 and 2 were determined for the first time by calculation of electronic circular dichroism (ECD) data. Among them, GKK1032B (10) showed significant cytotoxicity against human osteosarcoma cell line MG63 with an IC50 value of 3.49 μmol·L-1, and a primary mechanistic study revealed that it induced the apoptosis of MG63 cellsvia caspase pathway activation.
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Humans , Apoptosis , Bone Neoplasms , Caspases , Osteosarcoma/drug therapy , PenicilliumABSTRACT
Objective: To investigate the effect and its underlying molecular mechanisms of essential oil from Saussurea costus in esophageal cancer cell line Eca109. Methods: The chemical composition of essential oil from Saussurea costus was investigated by gas chromatography-mass spectrometry (GC-MS). The anti-proliferative, anti-migrative, and apoptotic effects of essential oil from Saussurea costus against Eca109 cells were analyzed. Moreover, the expression of proteins associated with cell cycle, metastasis, and apoptosis was determined. Results: GC-MS analysis showed that essential oil from Saussurea costus was predominantly comprised of sesquiterpenes. Saussurea costus essential oil inhibited the viability of Eca109 cells in a dose-and time-dependent manner with IC 50 values of (24.29±1.49), (19.16±2.27) and (6.97±0.86) μg/mL at 12, 24, and 48 h, respectively. The expression levels of target proteins in the cell cycle (phase G 1 /S), including cyclin D1, p21, and p53, were affected by Saussurea costus essential oil. The essential oil also downregulated the expression of metastasis-related proteins MMP-9 and MMP-2. Moreover, it induced apoptosis of Eca109 cells through the mitochondrial pathway, as well as inhibition of STAT3 phosphorylation. Conclusions: The essential oil from Saussurea costus exhibited anti-proliferative, anti-migrative, and apoptotic effects on Eca109 cells, and could be further explored as a potential anti-esophageal cancer agent.
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Abstract The aim of the present study was to evaluate the in vitro antiproliferative activity of ethanolic extract of leaves and fruits Citrus paradisi plant on HepG-2 liver cell lines by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-terazolium bromide) assay and to isolate and characterize the antiproliferative compounds by TLC (Thin layer chromatography) and FT-IR (Fourier transforms Infrared) spectroscopy. Qualitative phytochemical screening tests were performed to detect phytochemicals compounds from the crude extracts. Antioxidant activity of the plant extracts were characterized by using DPPH (2,2-Diphenyl-1-picrylhydrazyl) free radical scavenging method. The results showed that antioxidant activity using DPPH were found to be increased in a concentration dependent manner and decreased cell viability and cell growth inhibition in a dose dependent manner. The findings from this study indicated that fruit extract exhibited good antiproliferation and antioxidant potential. The seven functional groups of phytocompounds such as carboxylic acid, amine salt, aromatic compounds, cyclic alkene, aldehyde, fluoro compounds and alkene were detected by FT-IR which indicated that fruit extracts of Citrus paradisi possessed vast potential as a medicinal drug especially in liver cancer treatment.
Resumo O objetivo do presente estudo foi avaliar a atividade antiproliferativa in vitro do extrato etanólico de folhas e frutos da planta Citrus paradisi em linhagens de células hepáticas HepG-2 por MTT (3- (4, 5-dimetil-2-tiazolil) -2, Ensaio de brometo de 5-difenil-2H-terazólio) e isolar e caracterizar os compostos antiproliferativos por espectroscopia de TLC (cromatografia de camada fina) e FT-IR (infravermelho com transformadas de Fourier). Testes qualitativos de triagem fitoquímica foram realizados para detectar compostos fitoquímicos nos extratos brutos. A atividade antioxidante dos extratos vegetais foi caracterizada pelo método de eliminação de radicais livres DPPH (2,2-difenil-1-picrilhidrazil). Os resultados mostraram que a atividade antioxidante usando DPPH aumentou de uma maneira dependente da concentração e diminuiu a viabilidade celular e a inibição do crescimento celular de uma maneira dependente da dose. Os resultados deste estudo indicaram que o extrato de fruta exibiu bom potencial antiproliferação e antioxidante. Os sete grupos funcionais de fitocompostos, como ácido carboxílico, sal de amina, compostos aromáticos, alceno cíclico, aldeído, compostos de flúor e alceno, foram detectados por FT-IR, o que indicou que extratos de frutas de Citrus paradisi possuíam vasto potencial como medicamento, especialmente no tratamento de câncer do fígado.
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The aim of the present study was to evaluate the in vitro antiproliferative activity of ethanolic extract of leaves and fruits Citrus paradisi plant on HepG-2 liver cell lines by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2Hterazolium bromide) assay and to isolate and characterize the antiproliferative compounds by TLC (Thin layer chromatography) and FT-IR (Fourier transforms Infrared) spectroscopy. Qualitative phytochemical screening tests were performed to detect phytochemicals compounds from the crude extracts. Antioxidant activity of the plant extracts were characterized by using DPPH (2,2-Diphenyl-1-picrylhydrazyl) free radical scavenging method. The results showed that antioxidant activity using DPPH were found to be increased in a concentration dependent manner and decreased cell viability and cell growth inhibition in a dose dependent manner. The findings from this study indicated that fruit extract exhibited good antiproliferation and antioxidant potential. The seven functional groups of phytocompounds such as carboxylic acid, amine salt, aromatic compounds, cyclic alkene, aldehyde, fluoro compounds and alkene were detected by FT-IR which indicated that fruit extracts of Citrus paradisi possessed vast potential as a medicinal drug especially in liver cancer treatment.
O objetivo do presente estudo foi avaliar a atividade antiproliferativa in vitro do extrato etanólico de folhas e frutos da planta Citrus paradisi em linhagens de células hepáticas HepG-2 por MTT (3- (4, 5-dimetil-2-tiazolil) -2, Ensaio de brometo de 5-difenil-2H-terazólio) e isolar e caracterizar os compostos antiproliferativos por espectroscopia de TLC (cromatografia de camada fina) e FT-IR (infravermelho com transformadas de Fourier). Testes qualitativos de triagem fitoquímica foram realizados para detectar compostos fitoquímicos nos extratos brutos. A atividade antioxidante dos extratos vegetais foi caracterizada pelo método de eliminação de radicais livres DPPH (2,2-difenil-1-picrilhidrazil). Os resultados mostraram que a atividade antioxidante usando DPPH aumentou de uma maneira dependente da concentração e diminuiu a viabilidade celular e a inibição do crescimento celular de uma maneira dependente da dose. Os resultados deste estudo indicaram que o extrato de fruta exibiu bom potencial antiproliferação e antioxidante. Os sete grupos funcionais de fitocompostos, como ácido carboxílico, sal de amina, compostos aromáticos, alceno cíclico, aldeído, compostos de flúor e alceno, foram detectados por FT-IR, o que indicou que extratos de frutas de Citrus paradisi possuíam vasto potencial como medicamento, especialmente no tratamento de câncer do fígado.
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Humans , Citrus paradisi , Liver Neoplasms/drug therapy , Plant Extracts/pharmacology , Cell Line , Spectroscopy, Fourier Transform Infrared , Phytochemicals , AntioxidantsABSTRACT
Autophagy is a critical cellular homeostatic mechanism, and its dysfunction is linked to invasive breast carcinoma (BRCA). Recently, several omics methods have been applied to explore autophagic regulators in BRCA; however, more reliable and robust approaches for identifying crucial regulators and druggable targets remain to be discovered. Thus, we report here the results of multi-omics approaches to identify potential autophagic regulators in BRCA, including gene expression (EXP), DNA methylation (MET) and copy number alterations (CNAs) from The Cancer Genome Atlas (TCGA). Newly identified candidate genes, such as
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@#【Objective】To prepare rapamycin（RAPA）sustained-release film and to evaluate its dissolution.【Methods】RAPA sustained- release film was created by using polymer polyactioglyconic acid （PLGA），copolymer of polyactic acid（PLA）and polyglycolic acid（PGA）. Drug content of the sustained-release film was determined using specificity test，recovery，relative standard deviation（RSD）and stability test. Then，the dissolution of the sustained- release film was analyzed.【Results】The concentration of RAPA had a linear relationship with peak area，which ranged between 0.408 μg/mL and 40.8 μg/mL through the standard curve. The specificity test of the drug content determination indicated the excipient of the film and the solution with 0.3% sodium dodecyl sulfate（SDS）did not affect in determining the RAPA content. The recovery and RSD were excellent through drug content determination in blank films，which had three different levels of RAPA concentrations. The mean RAPA content of the sustained-release films was（112.6±10.1）μg（RSD 8.99%）through the drug content determination of the films，and the stability of RAPA with 0.3% SDS was good within 15 days. In addition，dissolution test of the sustained- release film indicated that the amount of drug release reached a high level and sustained up to 15 days.【Conclusion】 The RAPA sustained-release film with certain behavioral characteristic parameters had a stable drug content and favorable sustained-release property，and it may have certain application potential in anti-proliferation after glaucoma filtering surgery.
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Objective:To design and synthesize mono-carbonyl curcumin analogs with tumor suppressor activi- ty using substituted benzaldehyde and acetone as the raw material. Methods: Acetone and substituted benzaldehyde were used as starting compounds to synthesize the target compounds via aldol condensation reaction. The antiproliferation activity and apoptosis-inducing activity of target compounds against HepG2 cells were evaluated by MTT assay and flow cytometry, respectively. The medicinal chemical properties of the compounds were calculated online from the website: www.molinspiration.com. Results: Six compounds were synthesized and their structures were verified by the 1H NMR, 13C NMR and MS data. Compound 1 exhibited more potent antiproliferative and apoptosis-inducing activity against HepG2 cells than curcumin. The medicinal chemical properties of 1 were more consistent with Lipinski’s Rule of Five and it was ea sier for 1 to penetrate cell membranes than other compounds. Conclusion: The introduction of nitro group into the ortho-position of aromatic rings in mono-carbonyl curcumin analogs could enhance the anticancer activity, which is of great significance for the design and synthesis of mono-carbonyl curcumin analogs with better anticancer activity.
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Medicinal plants present a plausible source for anticancer agents. Combination of plant extracts and plant-derived compounds with the currently used cancer drugs has shown a marked improvement of the conventional drugs' efficacy and reduced toxicity. This study evaluated; phytochemical screening, antiproliferative activity and drug interaction potentials of Moringa oleifera and Indigofera arrecta leaf extracts with 5-fluoro uracil against selected cancer cell lines. Phytochemical screening was done using standard procedures. The common 3– (4, 5-dimethylthiazol-2-yr) -2, 5-diphenyltetrazolium (MTT) assay was used to determine the growth inhibitory potential of the extracts towards cancer cells. Drug interaction assays were done using constant ratio combination method. Alkaloids, terpenoids, tannins, flavonoids, glycosides, phenols and saponins were found to be present in the plant's extracts. M. oleifera and I. arrecta methanol-dichloromethane extracts had the highest activity compared to water extracts. All the extracts showed antiproliferative activities towards; HCC 1395 (breast), DU145 (prostate) and Hela (cervical) cancer cell lines. The extracts were not cytotoxic towards Vero cells (IC50>1000 µg/ml). I. arrecta and M. oleifera inhibited DU145 the most with IC50 values of 111.110 µg/ml and 66.290 µg/ml respectively. The plant extracts synergistically inhibited the growth of cancer cells (CI<1). Combination of the plant extracts and 5-Fluorouracil depicted that the concentration of the conventional drug could be reduced and yet achieve the same desired effect against cancerous cells (Dose reduction index (DRI) >1). Further studies to isolate the bioactive compounds and deduce the probable mechanisms of action are recommended.
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Aim To investigate the anti-proliferation effect of honokial on human colon cancer cells HCT116 and the possible mechanism. Methods Crystal stai-ning and flow cytometry assay were introduced to test the proliferation inhibitory and cell cycle arrested effect of honokial on HCT116 cells. Western blot was used to analyse the expression of PCNA and induction effect of apoptosis. Western blot and PCR assay were conducted to assess the change of BMP7. Via exogenous adenovi-ruses of BMP7 and its antibody combined with honoki-al,crystal violet staining and Western blot were used to analyse the effect on HCT116 cells. Results Honoki-al inhibited the growth of HCT116 in a time-and con- centration-dependent way,induced apoptosis and arres-ted at G2 phase; Western blot assay showed honokial up-regulated the level of PCNA and BMP7,and down-regulated the expression of Bcl-2; Western blot result also showed that exogenous adenoviruses of BMP7 en-hanced the effect of honokial on proliferation inhibition and apoptosis, while BMP7 antibody reversed such effects of honokial. Conclusion Honokial can inhibit the proliferation and induce apoptosis on HCT116 cells,which may be mediated by the up-regulation of BMP7.
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Aim To study the effect of tetrandrine ( Tet ) on proliferation of MCF-7 breast cancer cells and the possible mechanism underlying this biological process. Methods CCK-8, flow cytometric and Western blot were introduced to analyze the effect of Tet on proliferation and apoptosis in MCF-7 cells.Re-al-time PCR and/or Western blot assay were employed to detect the effect of Tet on expression of IGFBP-5 , p53 and MDM2.CCK-8 and recombinant adenovirus were utilized to determine the effect of IGFBP-5 on the proliferation inhibitory effect of Tet .Western blot assay was introduced to evaluate the effect of IGFBP-5 on p53 which was induced by Tet .Results Tet inhibited the proliferation , arrested cell cycle at G 1 phase and decreased the expression of PCNA concentration dependently in MCF-7 cells.Meanwhile, Tet increased the percentage of apoptotic cells , the level of Bad and reduced the level of Bcl-2.Tet increased the expres-sion of IGFBP-5 either mRNA or protein , over-expres-sion of IGFBP-5 enhanced the anti-proliferation activity of Tet in MCF-7 cells, but knockdown of IGFBP-5 at-tenuated this effect of Tet .Tet increased the level of p53 and decreased that of MDM2, and exogenous IG-FBP-5 enhanced the effect of Tet on p53 and MDM2, respectively .Conclusion Tet can inhibit the prolifer-ation of MCF-7 cells, and this activity is partly media-ted by increasing the function of p 53 signal , which may be triggered by the Tet-induced IGFBP-5.
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Objective To synthesize trifluoromethyl-substituted mono-carbonyl curcumin analogs and investigate their effects on the proliferation of human lung cancer cells A549 and NCI-H460. Methods Six mono-carbonyl curcumin analogs 1a-1f were synthesized via Aldol condensation using commercially available 2-trifluoromethyl benzadehyde and different ketones (actone, cyclopentanone, cyclohexanone, piperid-4-one, and 1-methylpiperid-4-one). MTT method was used to test the effect of 1a-1f on the proliferation of A549 and NCI-H460 cells. Results Compared with curcumin, most of the mono-carbonyl curcumin analogs 1a-1f exhibited anti-proliferation activity on the A549 cells. The values of IC50 were lower than that of curcumin (P＜0.01), and 1a and 1f showed much better activities both on the A549 and NCI-H460 cells, with their IC50 values were much lower than that of curcumin (P＜0.01).Among them, 1f showed better medicinal chemical properties, with the relative molecular mass less than 500, cLogP 5.25, and the polar surface area (PSA) 37.38, which was more better accorded with the Lipinski′s five rules. Conclusion Six mono-carbonyl curcumin analogs 1a-1f were synthesized, and 1f showed much better anti-proliferation activity on A549 and NCI-H460 cells.
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Objective Based on the previous research that the ethanolic extract from traditional Chinese medicine fructus forsythiae (Lianqiao) can obviously inhibit cancer cells in vitro, the article aimed to investigate the anti-proliferation effects of dammar-24-ene-3β-acetate-20S-ol (DM) extracted from fructus forsythiae on gastric cancer cells and its mechanism.Methods MTT assay was used to assess the anti-proliferation effects of DM on gastric cancer cells including SGC-7901, BGC-823, and MKN-45 in vitro.There were MKN-45 control group and its low dose and high dose groups, BGC-823 control group and its low dose and high dose groups, SGC-7901 control group and its low dose and high dose groups in the experiment.Flow cytometry was used to analyze the cell apoptosis rate.Cellquest software was used to analyze the results and record the ratio of cells at different cycles.DCFH-DA probe was applied to detect the ROS levels of blank control group, docetaxol group and DM group.The reaction system of microtubule assembly test was set with 10?mol/L docetaxol, 50 or 100 μmol/L DM final density and no medicine in blank control group.The readings of UV spectrophotometer were recorded.Microtubule assembly assay and microtubule immunofluorescence staining were applied to investigate the effects of DM on microtubule system.Results The inhibition ratio of 50 μg/L DM on the proliferation three gastric cell lines were all above 80%, with IC50s of MKN-45 11.72±1.35 μg/mL, BGC-823 17.19±0.82 μg/mL, SGC-7901 7.55±0.79 μg/mL.8 days′ low density culturing at 48 hours after 2 μg/mL DM treatment, compared with control group, the number of cell clones significantly reduced without much change in clone size, while 48 hours after 10 μg/mL DM treatment, besides a few clones of BGC-823, there were just several megascopic clones of SGC-7901 and MKN-45.In comparison with apoptotic cell ratio in MKN-45 control group[(21.1±2.5)%], its low dose group and high dose group resulted in significant rise of apoptotic cell ratio[(25.1±1.3)% and (55.2±2.3)%] (P0.05).In comparison with MKN-45 control group, the ratio of cells at S phase decreased in its low dose group[(14.5±2.7)% vs (12.3±3.3)%,P>0.05].In comparison with BGC-823 control group, the ratio of cells at S phase increased in its low dose group[(12.2±5.4)% vs (20.2±2.1)%,P<0.05].In comparison with SGC-7901 control group, the ratio of cells at S phase increased in its low dose group[(21.5±3.8)% vs (31.3±2.6)%,P<0.05].From the detection of intracellular active oxygen after DM treatment, dose-dependent ROS level increased in all three cell lines 48 hours after 10μg/mL and 50μg/mL DM treatment.From the results of microtubule immunofluorescence staining, 48 hours after the treatment of IC50 docetaxol and 10μg/mL DM, the fluorescence signals were in local concentration and disorder.Conclusion Dammar-24-ene-3β-acetate-20S-ol demonstrated anti-proliferation effects due to the apoptosis induced by cell cycle arrest at S phase.
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Euphorbia kansui is a commonly used traditional Chinese medicine for the treatment of edema, pleural effusion, and asthma, etc. According to the previous researches, terpenoids in E. kansui possess various biological activities, e.g., anti-virus, anti-allergy, antitumor effects. In this work, twenty five terpenoids were isolated from E. kansui, including thirteen ingenane- and eight jatrophane-type diterpenoids (with two new compounds, kansuinin P and Q) and four triterpenoids. Eighteen of them were analyzed by MTS assay for in vitro anticancer activity in five human cancer cell lines. Structure-activity relationship for 12 ingenane-type diterpenoids in colorectal cancer Colo205 cells were preliminary studied. Significant anti-proliferation activities were observed in human melanoma cells breast cancer MDA-MB-435 cells and Colo205 cells. More than half of the isolated ingenane-type diterpenoids showed inhibitory activities in MDA-MB-435 cells. Eight ingenane- and one jatrophane-type diterpenoids possessed much lower IC values in MDA-MB-435 cells than positive control staurosporine. Preliminary structure-activity relationship analysis showed that substituent on position 20 was important for the activity of ingenane-type diterpenoids in Colo205 cells and substituent on position 3 contributed more significant biological activity of the compounds than that on position 5 in both MDA-MB-435 and Colo205 cells.
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Humans , Cell Line, Tumor , Cell Proliferation , Drugs, Chinese Herbal , Chemistry , Pharmacology , Euphorbia , Chemistry , Molecular Structure , Neoplasms , Drug Therapy , Structure-Activity Relationship , Terpenes , Chemistry , PharmacologyABSTRACT
Aim To study the anti-proliferation effect of resveratrol (Res)and the role of Res-induced bone morphorgenetic protein 9 (BMP9 )in this process in colon cancer cells.Methods Crystal violet staining and flow cytomtry were introduced to assay the anti-proliferation effect of Res in LoVo cells.The effect of Res on apoptosis in LoVo cells was also detected with flow cytometry.Then,RT-PCR and Western blot assay were employed to unveil the effect of Res on the ex-pression of BMP9 .The effect of BMP9 on the anti-pro-liferation of Res in LoVo cells was analyzed with crystal violet staining and flow cytometry too.Finally,the effect of Res on the expression of ALK2 and ALK3 was assayed with RT-PCR,and the inhibitor of ALK2 and ALK3 was used to figure out the possible mechanism of BMP9 on Res-induced proliferation inhibition in LoVo cells.Results Res apparently inhibited the prolifera-tion,arrested the cell cycle at S phase in LoVo and in-creased the percentage of apopotic cells in LoVo cells. Res increased the expression of mRNA and protein of BMP9 concentration dependently. Exogenous ex-pressed-BMP9 enhanced the anti-proliferation and ap-optosis inducing effects of Res in LoVo cells, but BMP9 knockdown decreased these effects of Res.Al-though Res had no apparent effect on increasing the phosphorylation of Smad1/5/8,it increased the ex-pression of ALK2 and ALK3 .Inhibition of ALK2 and ALK3 decreased the anti-proliferation effect of Res partly in LoVo cells.Conclusion Res is potent to in-hibit the proliferation of LoVo cells,Which may be mediated by up-regulating the expression of BMP9 and its receptor at least.
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Objective To determine the cytotoxicity, reduction in nitric oxide production and anti-oxidative activity of the aqueous leaf extract from Tithonia diversifolia (T. diversifolia) in an in vitro model. Methods Leaves of T. diversifolia were collected from natural habitats and extracted with distilled water using the decoction method. The cytotoxic effect of the extract in terms of cell viability was determined using RAW264.7 cells and human peripheral blood mononuclear cells (PBMCs) via the mitochondrial respiration method using the MTT reagent. The effect of the extract on lipopolysaccharide (LPS)-induced nitric oxide production in RAW264.7 cells was measured using the Griess reagent. The chemical antioxidant was evaluated by ABTS- and DPPH-radical scavenging assays. Results The half-maximal cytotoxic concentration values were 145.87 μg/mL and 73.67 μg/mL for human PBMCs and RAW264.7 cells, respectively. In the presence of phytohemagglutinin-M, the IC
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Objective: To determine the cytotoxicity, reduction in nitric oxide production and anti-oxidative activity of the aqueous leaf extract from Tithonia diversifolia (T. diversifolia) in an in vitro model. Methods: Leaves of T. diversifolia were collected from natural habitats and extracted with distilled water using the decoction method. The cytotoxic effect of the extract in terms of cell viability was determined using RAW264.7 cells and human peripheral blood mononuclear cells (PBMCs) via the mitochondrial respiration method using the MTT reagent. The effect of the extract on lipopolysaccharide (LPS)-induced nitric oxide pro-duction in RAW264.7 cells was measured using the Griess reagent. The chemical anti-oxidant was evaluated by ABTS-and DPPH-radical scavenging assays. Results: The half-maximal cytotoxic concentration values were 145.87 mg/mL and 73.67 mg/mL for human PBMCs and RAW264.7 cells, respectively. In the presence of phytohemagglutinin-M, the IC50 on PBMCs proliferation was 4.42 mg/mL. The non-cytotoxic range of the extracts inhibited LPS-induced nitrite production in RAW264.7 cells with an IC50 value of 11.63 mg/mL. To determine the anti-oxidative properties, the N-acetyl cysteine equivalent antioxidant capacity of the extract was (32.62 ± 1.87) and (20.99 ± 2.79) mg N-acetyl cysteine/g extract, respectively determined by the ABTS-radical and DPPH-radical assay. However, the extract did not confer death protection in a hydrogen peroxide-induced RAW264.7 co-culturing model. Conclusions: Our study demonstrated the immunomodulation caused by the aqueous leaf extract of T. diversifolia, resulting from the inhibition of phytohemagglutinin-M-induced PBMCs proliferation and LPS-induced nitric oxide production in RAW264.7 macrophages. Although the anti-oxidative activity was presented in the chemical-based anti-oxidant assay, the extract cannot protect cell death from stress conditions.
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Objective To study the effect of mitomycin C(MMC)‐chitosan (CS)sustained release microspheres on the scar‐ring prevention of the filtering passage during glaucoma filtering surgery .Methods Filtering surgery model was established on 96 eyes of 48 healthy adult New Zealand white rabbits which were divided into 4 groups ,24 eyes in each group .The right eyes implan‐ted MMC‐CS microspheres in 24 rabbits(group A) ,whereas that of the fellow eyes implanted empty CS microspheres(group B) , and in other 24 rabbits which right eyes only used 0 .20 mg/mL MMC cotton(group C) ,on the opposite sides with no adjunctive treatment(group D) .Intraocular pressure ,filtering bleb ,anterior chamber flare ,complications and the numbers of corneal endotheli‐al cells were observed after surgery .Rabbits were killed on the 7th ,14th and 21th day postoperatively in batch and histopathological examination was carried out .Results (1)the intraocular pressure:group A can maintain the low level of intraocular pressure for a long time ,followed by group C ,group B and group D ,and difference between groups was statistically significant (P0 .05) .(5)Histopathologic findings :group A had bet‐ter conjunctival epithelial integrity ,subconjunctival fibroblast less than other groups ,filtration had no obvious inflammation infiltra‐tion around the mouth area ,but also had fewer new collagen fibers .Conclusion MMC‐CS sustained release microspheres is a safe and effective treatment to inhibit inflammatory cells activity and fibroblast activity in surgery sites ,and can significantly improve outcome of filtration surgery .
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Objective To construct a eukaryotic expression system of IFN-λ,examine the expression of IFN-λand evaluate its bio-functions including anti-proliferation and anti-viral activity.Methods The genes of human IFN-λ1 /2 (hIFN-λ1 /2)were cloned from the mRNA of poly I∶C treated HuH-7 cells.The PCR product was examined with DNA sequencing.The genes of IFN-λ1 /2 were sub-cloned into pcDNA3 vector.The correct insertion of the gene IFN-λ1 /2 was identified with enzyme digestion.The constructed pcDNA3-IFN-λ1 /2 plasmids were transfected into COS-7 cells and IFN-λ1 /2 protein was checked in the supernatant and lysis of transfected cells using Western blotting analysis.The human esophageal carcinoma YES5 and T.Tn cells were treated with the IFN-λ1 /2 from the transfected cells and the proliferation of carcinoma cells were measured with CCK-8 kit.In the treated carcinoma cells,the apoptosis and antivirus related molecules such as caspase-3,ISG15 and MxA was analyzed with Western blotting or Quantitative real time PCR.Results The sequence of hIFN-λ1 /2 fragment matched that of the gene bank and the gene of the cytokines was inserted into pcDNA3 vector correctly.With Western blotting analysis,IFN-λ1 /2 protein was detected in the pcDNA3-IFN-λ1 /2 transfected COS-7 cells.The IFN-λ1 /2 from the transfected COS-7 cells inhibited the growth of YES5 and T.Tn cells, activated apoptosis related caspase-3,and up-regulated the anti-virus gene expression of ISG15 and MxA.Conclusion COS-7 cells can express IFN-λ1 /2 after transfection with pcDNA3-IFN-λ1 /2,suggesting that eukaryotic expression system of IFN-λis established.IFN-λ1 /2 from the system can perform bio-functions,such as proliferation inhibition,apoptosis induction and anti-viral gene up-regulation,which indicates that the system can contribute to further investigations of IFN-λbio-activity and its clinical application.
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The triggering receptor expressed on myeloid cells (TREM) family, which is abundantly expressed in myeloid lineage cells, plays a pivotal role in innate and adaptive immune response. In this study, we aimed to identify a novel receptor expressed on hematopoietic stem cells (HSCs) by using in silico bioinformatics and to characterize the identified receptor. We thus found the TREM-like transcript (TLT)-6, a new member of TREM family. TLT-6 has a single immunoglobulin domain in the extracellular region and a long cytoplasmic region containing 2 immunoreceptor tyrosine-based inhibitory motif-like domains. TLT-6 transcript was expressed in HSCs, monocytes and macrophages. TLT-6 protein was up-regulated on the surface of bone marrow-derived and peritoneal macrophages by lipopolysaccharide stimulation. TLT-6 exerted anti-proliferative effects in macrophages. Our results demonstrate that TLT-6 may regulate the activation and proliferation of macrophages.
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Humans , Adaptive Immunity , Computational Biology , Computer Simulation , Cytoplasm , Hematopoietic Stem Cells , Immunoglobulins , Macrophages , Macrophages, Peritoneal , Monocytes , Myeloid CellsABSTRACT
AIM@#To study the chemical constituents from the roots of Buleurum bicaule Helm (Apiaceae).@*METHOD@#Silica gel, Sephadex LH-20, MPLC Rp-C18 column chromatography, and HPLC were used for isolation of compounds. The structures were elucidated on the basis of 1D- and 2D-NMR technology and HRESI-MS. Compounds were evaluated in vitro for their inhibitory ability against the proliferation of rat mesangial cells by the MTT method.@*RESULTS@#Twelve compounds were isolated, and their structures were identified on the basis of their spectroscopic and physico-chemical properties as 13, 28-epoxy-olean-11-en-3-one (1), saikogenin E (2), saikogenin G (3), 11α-methoxy-3β, 16β, 23, 28-tetrahydroxyolean-12-ene (4), saikogenin D (5), prosaikogenin F (6), prosaikogenin A (7), prosaikogenin G (8), prosaikogenin D (9), laccaic acid (10b), methyl gallate (11), and ethyl gallate (12). Compounds 1, 2, 7, 8, and 10 were observed to have inhibitory activity against mesangial cell proliferationin to different degrees.@*CONCLUSION@#Compound 1, 8, and 10 exhibit significant inhibitory effects on rat mesangial cell proliferation induced by Ang II.