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1.
Food Res Int ; 183: 114202, 2024 May.
Article in English | MEDLINE | ID: mdl-38760133

ABSTRACT

Pixian broad bean paste is a renowned fermented seasoning. The fermentation of broad bean is the most important process of Pixian broad bean paste. To enhance the flavor of tank-fermented broad bean paste, salt-tolerant Bacillus amyloliquefaciens strain was inoculated, resulting in an increase in total amount of volatile compounds, potentially leading to different flavor characteristics. To investigate the fermentation mechanism, monoculture simulated fermentation systems were designed. Metabolomics and transcriptomics were used to explore Bacillus amyloliquefaciens' transcriptional response to salt stress and potential aroma production mechanisms. The results highlighted different metabolite profiles under salt stress, and the crucial roles of energy metabolism, amino acid metabolism, reaction system, transportation system in Bacillus amyloliquefaciens' hypersaline stress response. This study provides a scientific basis for the industrial application of Bacillus amyloliquefaciens and new insights into addressing the challenges of poor flavor quality in tank fermentation products.


Subject(s)
Bacillus amyloliquefaciens , Fermentation , Metabolomics , Bacillus amyloliquefaciens/metabolism , Bacillus amyloliquefaciens/genetics , Transcriptome , Food Microbiology , Fermented Foods/microbiology , Volatile Organic Compounds/analysis , Volatile Organic Compounds/metabolism , Gene Expression Profiling , Taste , Fabaceae/microbiology
2.
Food Chem X ; 17: 100556, 2023 Mar 30.
Article in English | MEDLINE | ID: mdl-36845488

ABSTRACT

With the advancement of industrialization, tank fermentation technology is promising for Pixian broad bean paste. This study identified and analyzed the general physicochemical factors and volatile metabolites of fermented broad beans in a thermostatic fermenter. Headspace solid-phase microextraction (HS-SPME)-two-dimensional gas chromatography-mass spectrometry (GC × GC-MS) was applied to detect the volatile compounds in fermented broad beans, while metabolomics was used to explore their physicochemical characteristics and analyze the possible metabolic mechanism. A total of 184 different metabolites were detected, including 36 alcohols, 29 aldehydes, 26 esters, 21 ketones, 14 acids, 14 aromatic compounds, ten heterocycles, nine phenols, nine organonitrogen compounds, seven hydrocarbons, two ethers, and seven other types, which were annotated to various branch metabolic pathways of carbohydrate and amino acid metabolism. This study provides references for subsequent functional microorganism mining to improve the quality of the tank-fermented broad beans and upgrade the Pixian broad bean paste industry.

3.
J Agric Food Chem ; 71(11): 4706-4716, 2023 Mar 22.
Article in English | MEDLINE | ID: mdl-36814172

ABSTRACT

To understand the umami taste of fermented broad bean paste (FBBP) and explore the umami mechanism, eight peptides (PKALSAFK, NKHGSGK, SADETPR, EIKKAALDANEK, DALAHK, LDDGR, and GHENQR) were separated and identified via ultrafiltration, RP-HPLC, and UPLC-QTOF-MS/MS methods. Sensory experiments suggested that eight novel peptides showed umami/umami-enhancing and salt-enhancing functions. Significantly, the threshold of EIKKAALDANEK in aqueous solution exceeded that of most umami peptides reported in the past 5 years. The omission test further confirmed that umami peptides contributed to the umami taste of FBBP. Molecular docking results inferred that all peptides easily bind with Ser, Glu, His, and Asp residues in T1R3 through hydrogen bonds and electrostatic interactions. The aromatic interaction, hydrogen bond, hydrophilicity, and solvent-accessible surface (SAS) were the main interaction forces. This work may contribute to revealing the secret of the umami taste of FBBP and lay the groundwork for the efficient screening of umami peptides.


Subject(s)
Tandem Mass Spectrometry , Taste , Molecular Docking Simulation , Peptides/chemistry , Chromatography, High Pressure Liquid , Receptors, G-Protein-Coupled/metabolism
4.
Food Sci Nutr ; 10(8): 2760-2772, 2022 Aug.
Article in English | MEDLINE | ID: mdl-35959259

ABSTRACT

Broad bean fermentation is of vital importance in PixianDouban (PXDB) production, as well as a key process for microorganisms to degrade protein, which lays the foundation for the formation of PXDB flavor. In this study, two fungi and bacteria were screened, and their morphology, molecular biology, growth, and enzyme production characteristics were analyzed, and then they were applied to the broad bean fermentation simulation system. The protein, peptide, amino acid, amino nitrogen, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system were evaluated. The results showed that the four microorganisms were Aspergillus oryzae, Aspergillus jensenii, Staphylococcus gallinarum, and Enterobacter hormaeche. Aspergillus oryzae had the highest protease activity at pH 7.0, while the other three strains had better enzyme activity stability under neutral acidic conditions. And the total protein (F1 and F2 were 18.32 g/100 g, 19.15 g/100 g, respectively), peptides (11.79 ± 0.04 mg/g and 12.06 ± 0.04 mg/g), and amino acids (55.12 ± 2.78 mg/g and 54.11 ± 1.97 mg/g) of the fungus experimental groups (F) were higher than the bacterial experimental groups (B). In addition, the enzyme system produced by fungi exhibited a stronger ability for albumin (20 kDa) and glutenin (<30 kDa) deterioration in neutral conditions, while the bacterial enzyme system was more efficient in degrading albumin (<30 kDa) and glutenin (20-30 kDa) in acidic conditions, as indicated by SDS-PAGE. These findings showed that both bacteria and fungi played an important role in the degradation of protein in different fermentation stages of broad bean fermentation. Practical applications: There is a lack of comprehensive understanding of the protein composition and protein degradation mechanism of broad beans in the fermentation stage of PXDB. This research work explored the differences in the degradation of PXDB fermented protein by different microorganisms, and provided a theoretical basis for optimizing the production of PXDB and improving the quality of PXDB.

5.
Food Funct ; 13(16): 8730-8740, 2022 Aug 15.
Article in English | MEDLINE | ID: mdl-35917196

ABSTRACT

Pixian broad-bean paste (PBBP) is a famous fermented condiment in China, which may produce abundant flavor peptides during the fermentation process. Herein, the flavor peptides from fermented broad-bean (FB) were separated and identified via ethanol precipitation, macroporous resin, and ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) methods. The results showed that there were fifteen novel flavor peptides in FB. Among them, six peptides (i.e. ALDELGT, AELTPEP, SAALQAG, SFEAVEAAPT, EQDNDGNNIFSGFKR, QTFNTEEDTAK, and DAPASGN) were perceived umami with threshold values ranging from 0.76 to 1.84 mmol L-1. And they had both salty-enhancing and umami-enhancing abilities. Meanwhile, novel peptides (i.e. GGLRIINPEGQQ, SIITPPERQ, GGLSIITPPERQA, GGLRIINPEG, SIITPPER, RIINPEGQQ, DALNVNRHIV, and LPKILLLQLV) were perceived bitter with threshold values ranging from 0.56 to 2.89 mmol L-1. And the monosodium glutamate (MSG) solution had a certain masking effect on bitter peptides. GFSSEFLA showed a strong sweet taste. From peptide sequence analysis, it was found that Aspergillus oryzae and broad-bean were important taste-active sources of the flavor peptides in FB. Besides, the synthetic peptides showed a much better sensory taste than the mixtures of the corresponding amino acids. It indicated that peptides might play an important role in deciding the taste of FB. Significantly, it is the first time that the isolated flavor peptides were identified from FB.


Subject(s)
Fabaceae , Vicia faba , Peptides/analysis , Tandem Mass Spectrometry , Taste
6.
RSC Adv ; 11(53): 33759-33769, 2021 Oct 08.
Article in English | MEDLINE | ID: mdl-35497520

ABSTRACT

Lung cancer is the leading cause of cancer-related deaths worldwide. Early diagnosis and treatment is critical to improving the 5 year survival rate of lung cancer. The identification of new options for early-stage diagnosis and therapy of lung cancer still represents a crucial challenge. Therefore, a new diagnostic method is urgently needed. In this study, we used a new modified SELEX, called serum-SELEX, to isolate aptamers that can specifically bind lung cancer serum, without any prior knowledge of their target. Among the obtained candidate aptamer sequences, Ap-LC-19 was identified as the optimal aptamer probe with the lowest dissociation constant (K d) value of 15 ± 8.6 nM and higher affinity assessed by qPCR. Furthermore, this molecule could be a suitable aptamer for lung cancer serum and could be used as a recognition element in aptamer-based biosensors for efficient early diagnosis of lung cancer or as an innovative tool for targeted therapy. In addition, we performed MALDI-TOF MS followed by secondary peptide sequencing MS analysis for the identification of the aptamer targeted proteins. CLEC3B could be useful biomarkers for early detection of lung cancer and in monitoring its evolution.

7.
J Cancer ; 11(21): 6356-6364, 2020.
Article in English | MEDLINE | ID: mdl-33033519

ABSTRACT

Radiotherapy is frequently applied for clinically localized prostate cancer while its efficacy could be significantly hindered by radioresistance. MicroRNAs (miRNAs) are important regulators in mediating cellular responses to ionizing radiation (IR), and strongly associate with radiosensitivity in many cancers. In this study, enhancement of radiosensitivity by miR-29b-3p was demonstrated in prostate cancer cell line LNCaP in vitro. Results showed that miR-29b-3p expression was significantly upregulated in response to IR from both X-rays and carbon ion irradiations. Knockdown of miR-29b-3p resulted in radioresistance while overexpression of miR-29b-3p led to increased radiosensitivity (showing reduced cell viability, suppressed cell proliferation and decreased colony formation). In addition, miR-29b-3p was found to directly target Wnt1-inducible-signaling protein 1 (WISP1). Inhibition of WISP1 facilitated the mitochondrial apoptosis pathway through suppressing Bcl-XL expression while activating caspase-3 and poly (ADP-ribose) polymerase (PARP). The results indicated that miR-29b-3p was a radiosensitizing miRNAs and could enhance radiosensitivity of LNCaP cells by targeting WISP1. These findings suggested a novel treatment to overcome radioresistance in prostate cancer patients, especially those with higher levels of the WISP1 expression.

8.
Bioorg Med Chem ; 27(5): 851-858, 2019 03 01.
Article in English | MEDLINE | ID: mdl-30723042

ABSTRACT

Fluorescein diester which is conjugated with cell membrane permeable Arg9 peptide was proposed as probe for ester prodrug stability and drug release study in living cells. α-Amino protected d-Val and l-Ala which bear differently hindered side chains were used to afford model diesters of 5-maleimide-fluorescein. Such fluorescein diesters were further conjugated with a Cys containing cell membrane permeable Arg9 peptide via thiol-ene crosslink reaction. The resulted conjugates of fluorescein diester and Arg9 peptide were purified with HPLC and characterized with MALDI-TOF MS. Upon incubation with cultured cells, the fluorescein diesters were delivered into the cells, the following hydrolysis of fluorescein diesters and release of fluorescein inside living cells were observed by monitoring the fluorescence accumulation. Fluorescence microscopic imaging studies of HeLa cells treated with fluorescein l-Ala diester show strong fluorescence accumulation in 30 min indicating fast hydrolysis of fluorescein diester and fluorescein release; in contrast d-Val diester remains stable inside cells evidenced by margin fluorescence formation. Further flowcytometry studies on the fluorescein diester-Arg9 conjugate treated cells show that the hydrolysis t1/2 for l-Ala diester is 15 min. The results also show that Arg9 peptide not only transports the ester probes into cell efficiently but also can retain and concentrate hydrolytic product fluorescein inside cells so that the accumulated fluorescence can be accurately quantified. This fluorogenic probe approach provides feasible applications in dynamic studies on ester prodrug hydrolysis and release, facilitating screening and optimization of prodrug structures in living cell settings.


Subject(s)
Cell-Penetrating Peptides/chemistry , Esters/chemistry , Fluoresceins/chemistry , Fluorescent Dyes/chemistry , Prodrugs/chemistry , Cell-Penetrating Peptides/chemical synthesis , Cell-Penetrating Peptides/metabolism , Drug Liberation , Esters/chemical synthesis , Esters/metabolism , Flow Cytometry , Fluoresceins/chemical synthesis , Fluoresceins/metabolism , Fluorescence , Fluorescent Dyes/chemical synthesis , Fluorescent Dyes/metabolism , HeLa Cells , Humans , Hydrolysis , Microscopy, Fluorescence , Prodrugs/chemical synthesis , Prodrugs/metabolism , Proof of Concept Study
9.
Toxicology ; 417: 35-41, 2019 04 01.
Article in English | MEDLINE | ID: mdl-30779955

ABSTRACT

In spite of carbon ion radiotherapy is a talented modality for malignant tumor patients, the radiation damage of normal tissues adjacent to tumor and the dysfunction of immune system limits therapeutic gain. Protecting immune system against carbon ion radiation-caused damage has the possibility to improve cancer treatment, but it is uncertain whether conventional radioprotective agents play a role in carbon ion radiation. To certify carbon ion caused immune dysfunction and assess the radioprotective effect of melatonin on immune system, animal experiments were performed in radiosensitive BALB/C mice. Here, we observed the bodyweight loss, death and apoptosis, abnormal T-cell distributions in immune system in carbon ion radiated mice. Pretreatment with melatonin could increase the index of thymus and spleen, reduce cell apoptosis in thymus and spleen, and attenuate the carbon ion radiation-caused imbalance of T lymphocytes and disorder of cytokines. These results suggest that melatonin can act as an effective protector against carbon ion radiation-caused immune dysfunction. Furthermore, we also found melatonin restored the activity of the antioxidant enzymes and reduced the level of lipid peroxidation in serum. These data have provided baseline information both for radiation workers and cancer patients to use melatonin as a radioprotector during the carbon ion radiation treatment.


Subject(s)
Heavy Ion Radiotherapy/adverse effects , Immunity, Cellular/drug effects , Immunity, Cellular/radiation effects , Melatonin/pharmacology , Radiation-Protective Agents/pharmacology , Animals , Apoptosis/drug effects , Apoptosis/immunology , Apoptosis/radiation effects , Dose-Response Relationship, Drug , Immunity, Cellular/immunology , Lipid Peroxidation/drug effects , Lipid Peroxidation/immunology , Lipid Peroxidation/radiation effects , Male , Mice , Mice, Inbred BALB C , Oxidative Stress/drug effects , Oxidative Stress/immunology , Oxidative Stress/radiation effects
10.
RSC Adv ; 9(27): 15513-15520, 2019 May 14.
Article in English | MEDLINE | ID: mdl-35514829

ABSTRACT

Neuron-specific enolase (NSE) is one of the most commonly used serum tumor biomarker in clinical practice for small cell lung cancer screening, early diagnosis, evaluation of therapeutic efficacy and prognosis. In this study, we obtained DNA aptamers with great affinity and selectivity to NSE via subtractive SELEX approach. After 10 rounds, three candidate aptamers were successfully selected and identified. Their affinities were measured by surface plasmon resonance. Apt-5 aptamer with high binding affinity and good specificity were obtained, which had the dissociation constant (K D) values of 12.26 nM. In addition, electrophoretic mobility shift assay (EMSA) experiment also further indicated that the Apt-5 had a highly specific affinity to NSE without binding to HSA. The circular dichroism (CD) analysis revealed that the three aptamers formed stable B-form, stem-loop conformations. The selected aptamers were used to construct a chemiluminescence (CL) aptasensor biosensing platform to detect NSE from actual serum samples. Experimental results confirmed that the CL immunosensing platform had good sensitivity with detection limits of 1-100 ng mL-1. The results demonstrated that our obtained the Apt-5 aptsensor was highly specific in the detection of NSE in serum samples. The detection limit was 0.1 ng mL-1, which was lower than the 0.25 ng mL-1 limit of the ELISA used at the hospital. Moreover, the aptasensor can contribute to better detection of small cell lung cancer (SCLC).

11.
RSC Adv ; 9(2): 950-957, 2019 Jan 02.
Article in English | MEDLINE | ID: mdl-35517584

ABSTRACT

Gastric cancer is still among the leading causes of cancer deaths worldwide. Despite the improvements in diagnostic methods, the status of early detection has not been achieved so far. Early diagnosis of gastric cancer may significantly improve the cure rate of patients. Therefore, a new diagnostic method is needed. In this study, subtractive SELEX was performed to screen gastric cancer serum-specific DNA aptamers by using gastric cancer serum and normal serum as the target and negative serum, respectively. Four highly specific aptamers generated for gastric cancer serum, Seq-3, Seq-6, Seq-19 and Seq-54, were developed using whole-serum subtractive SELEX technology with K d of 128 ± 26.3 nM, 149 ± 23.6 nM, 232 ± 44.2 nM, 202 ± 25.6 nM, respectively. These generated aptamers showed higher specificities toward their target serum by differentiating normal serum but closely related other cancer serums. The selected four high affinity DNA aptamers were further applied to the development based on qPCR method for the early detection of gastric cancer. In addition, we performed MALDI-TOF MS followed by secondary peptide sequencing MS analysis for the identification of the aptamer binding proteins. Among these potential biomarkers, APOA1, APOA4, PARD3, Importin subunit alpha-1 showed a relatively high score probability. Therefore, these four ssDNA aptamers generated in our study could be a promising molecular probe for gastric cancer diagnosis.

12.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-793189

ABSTRACT

@# miR-29b是最近生物医学界所关注的研究热点之一,尤其在人类癌症中。越来越多的研究发现,miR-29b在多种癌症 中异常表达,与肿瘤细胞增殖、分化、凋亡、侵袭、转移及药物耐受相关,有望成为癌症的新型诊断标志物及治疗靶点。本文重点 就miR-29b在人类癌症中的表达、作用及其调控机制和临床应用的研究进展进行综述,以促进miR-29b在临床诊断和治疗中的转 化应用。

13.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi ; 35(6): 964-969, 2018 12 25.
Article in Chinese | MEDLINE | ID: mdl-30583324

ABSTRACT

Nucleic acid aptamer is an oligonucleotide sequence screened by the exponential enrichment ligand system evolution technology (SELEX). Previous studies have shown that nucleic acid aptamer has a good application prospect in tumor diagnosis and treatment. Therefore, we reviewed the selection and identification of nucleic acid aptamer of lung cancer cells in recent years, and discussed the effect of aptamer as targeting drugs and targeting vectors on the diagnosis of tumors, which provide a new idea for early diagnosis and treatment of tumor.

14.
ACS Appl Mater Interfaces ; 9(20): 16756-16766, 2017 May 24.
Article in English | MEDLINE | ID: mdl-28472883

ABSTRACT

Inspired by marine mussel adhesive proteins, polymers with catechol side groups have been extensively explored in industrial and academic research. Here, Pluronic L-31 alcoholate ions were used as the initiator to prepare a series of polypeptide-Pluronic-polypeptide triblock copolymers via ring-opening polymerization of l-DOPA-N-carboxyanhydride (DOPA-NCA), l-arginine-NCA (Arg-NCA), l-cysteine-NCA (Cys-NCA), and ε-N-acryloyl lysine-NCA (Ac-Lys-NCA). These copolymers demonstrated good biodegradability, biocompatibility, and thermoresponsive properties. Adhesion tests using porcine skin and bone as adherends demonstrated lap-shear adhesion strengths up to 106 kPa and tensile adhesion strengths up to 675 kPa. The antibleeding activity and tissue adhesive ability were evaluated using a rat model. These polypeptide-Pluronic copolymer glues showed superior hemostatic properties and superior effects in wound healing and osteotomy gaps. Complete healing of skin incisions and remodeling of osteotomy gaps were observed in all rats after 14 and 60 days, respectively. These copolymers have potential uses as tissue adhesives, antibleeding, and tissue engineering materials.


Subject(s)
Peptides/chemistry , Adhesives , Animals , Biocompatible Materials , Hemostasis , Poloxamer , Rats , Swine , Tissue Adhesives
15.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 25(2): 444-448, 2017 Apr.
Article in Chinese | MEDLINE | ID: mdl-28446290

ABSTRACT

OBJECTIVE: To investigate the relationship of gene polymorphisms of inflammattion related cytokines with incidence of diffuse large B-cell lymphoma(DLBCL) in Gansu Han population. METHODS: The gene polymorphism of inflammation-related cytokines were detected by high-resolution melting(HRM) curve. RESULTS: The homozygous CC genotype carrying IL-1RA rs4251961 gene locus was related with the risk of DLBCL in comparison with homozygous TT, the OR was 0.83 of homozygous CC, 95% CI=0.697-0.997,P<0.05), while the C allele of IL-1RA rs4251961 gene locus significantly correlated with the high incidence of diffuse large B-cell lymphoma compared with T allele(OR=8.83, 95% CI=1.909-40.813,P<0.01). CONCLUSION: The minor allele C of IL-1RA rs4251961 gene locus significantly relates with the susceptibility to DLBCL.


Subject(s)
Genetic Predisposition to Disease , Lymphoma, Large B-Cell, Diffuse/genetics , Cytokines , Genotype , Humans , Inflammation/genetics , Lymphoma, Large B-Cell, Diffuse/drug therapy , Polymorphism, Genetic , Polymorphism, Single Nucleotide
16.
Biomed Pharmacother ; 85: 763-771, 2017 Jan.
Article in English | MEDLINE | ID: mdl-27923690

ABSTRACT

Dicranostiga Leptodu (Maxim.) fedde (DLF), a poppy plant, has been reported have many benefits and medicinal properties, including free radicals scavenging and detoxifying. However, the protective effect of DLF extracts against carbon tetrachloride (CCl4)-induced damage in mice liver has not been elucidated. Here, we demonstrated that DLF extracts attenuated CCl4-induced liver damage in mice through increasing anti-oxidative enzyme activity to improve mitochondrial function. In this study, the mice liver damage evoked by CCl4 was marked by morphology changes, significant rise in lipid peroxidation, as well as alterations of mitochondrial respiratory function. Interestingly, pretreatment with DLF extracts attenuated CCl4-induced morphological damage and increasing of lipid peroxidation in mice liver. Additionally, DLF extracts improved mitochondrial function by preventing the disruption of respiratory chain and suppression of mitochondrial Na+K+-ATPase and Ca2+-ATPase activity. Furthermore, administration with DLF extracts elevated superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) levels and maintained the balance of redox status. This results showed that toxic protection effect of DLF extracts on mice liver is mediated by improving mitochondrial respiratory function and keeping the balance of redox status, which suggesting that DLF extracts could be used as potential toxic protection agent for the liver against hepatotoxic agent.


Subject(s)
Antioxidants/metabolism , Carbon Tetrachloride Poisoning/drug therapy , Chemical and Drug Induced Liver Injury/prevention & control , Mitochondria/drug effects , Papaveraceae/chemistry , Plant Extracts/pharmacology , Animals , Calcium-Transporting ATPases/genetics , Calcium-Transporting ATPases/metabolism , Chemical and Drug Induced Liver Injury/pathology , Gene Expression Regulation, Enzymologic/drug effects , Lethal Dose 50 , Mice , Mitochondria/metabolism , Oxidative Stress/drug effects , Plant Extracts/chemistry , Sodium-Potassium-Exchanging ATPase/genetics , Sodium-Potassium-Exchanging ATPase/metabolism
17.
J Biomater Sci Polym Ed ; 24(12): 1472-83, 2013.
Article in English | MEDLINE | ID: mdl-23829459

ABSTRACT

Human hepatocellular carcinoma (HCC) is one of the major causes of death worldwide. To investigate the relative importance of active and passive targeting strategies, the synthesis, characterization, in vitro uptake, and in vivo biodistribution of specific sulfapyridine HPMA (HPMA: N-(2-hydroxypropyl methacrylamide)) copolymer (sulfapyridine: SPD) conjugates, nonspecific HPMA copolymer conjugates, and DTPA are described in this study. The poly(HPMA)-SPD-DTPA (DTPA: diethylenetriaminepentaacetic acid), poly(HPMA)-DTPA, and DTPA conjugates were radiolabeled with the radionuclide (99m)Tc and tested for uptake by cultured H22 cells. The cellular accumulation of poly(HPMA)-SPD-DTPA-(99m)Tc complex was found to be time-dependent. The poly(HPMA)-SPD-DTPA-(99m)Tc tracer exhibited rapid uptake kinetics in cell culture with a t(1/2) of ~5 min. The uptake of poly(HPMA)-SPD-DTPA-(99m)Tc was significantly higher than that of poly(HPMA)-DTPA-(99m)Tc, indicating that the uptake of the poly(HPMA)-SPD-DTPA-(99m)T was active binding. The uptake of poly(HPMA)-DTPA-(99m)Tc was significantly higher than that of DTPA-(99m)Tc, suggesting that the uptake of the poly(HPMA)-DTPA-(99m)T was passive binding. Twenty-four hour necropsy data in the hepatocellular carcinoma tumor model showed significantly higher (p < 0.001) tumor localization for poly(HPMA)-SPD-DTPA-(99m)Tc (4.98 ± 0.48%ID/g [percentage injected dose per gram tissue]) compared with poly(HPMA)-DTPA-(99m)Tc (2.69 ± 0.15% ID/g) and DTPA-(99m)Tc (0.83 ± 0.03%ID/g). Moreover, higher T/B for poly(HPMA)-SPD-DTPA-(99m)Tc indicated reduced extravazation of the targeted polymeric conjugates in normal tissues. Specific molecular targeting and nonspecific vascular permeability are both significant in the relative tumor localization of poly(HPMA)-SPD-DTPA-(99m)Tc. Extravascular leak in nonspecific organs appears to be a major factor in reducing the T/B for the sulfapyridine molecules. Thus, the poly(HPMA)-SPD-DTPA is expected to be used as the potential macromolecular targeting carrier for hepatoma carcinoma in mice.


Subject(s)
Acrylamides/chemical synthesis , Pentetic Acid/analogs & derivatives , Polymethacrylic Acids/chemical synthesis , Polymethacrylic Acids/pharmacokinetics , Radiopharmaceuticals/chemical synthesis , Acrylamides/chemistry , Acrylamides/pharmacokinetics , Animals , Carcinoma, Hepatocellular/diagnostic imaging , Carcinoma, Hepatocellular/metabolism , Cell Line, Tumor , Drug Delivery Systems , Liver Neoplasms/diagnostic imaging , Liver Neoplasms/metabolism , Male , Mice , Neoplasm Transplantation , Pentetic Acid/chemical synthesis , Pentetic Acid/chemistry , Pentetic Acid/pharmacokinetics , Polymethacrylic Acids/chemistry , Radionuclide Imaging , Radiopharmaceuticals/chemistry , Radiopharmaceuticals/pharmacokinetics , Technetium , Tissue Distribution
18.
Zhonghua Zhong Liu Za Zhi ; 34(11): 826-30, 2012 Nov.
Article in Chinese | MEDLINE | ID: mdl-23291130

ABSTRACT

OBJECTIVE: To explore the pathogenesis of tumors by blocking the normal differentiation process of stem cells. METHODS: Bone marrow mesenchymal stem cells (BMSCs) from rats were isolated, cultured and purified by whole bone marrow adherence method. The rat BMSCs were induced to differentiate into adipocytes with dexamethasone, insulin and indomethacin. Blockage of the differentiation process was induced by 3-methylcholanthrene (3-MC). RESULTS: The differentiation experiment showed that at 30 days after the induction, oil red O staining-positive cells occurred with increased intracytolasmic lipid droplets, characteristic for adipocytes. The differentiation blockage experiment showed that at 30 days after induction, the deposits of oil red O staining-cytoplasmic lipid droplets was significantly reduced, indicating that the blocked cells were adipocytes, but not fully differentiated. Morphological identification showed that cell contact inhibition disappeared, abnormal cell nuclei, increased number of micronucleus aberration and karyotype abnormalities, indicating that malignant transformation of the stem cells occurred after the differentiation blockage. CONCLUSIONS: The results of this study show a blockage of the differentiation of that stem cells at the intermediate phase, and a tendency of malignant transformation of the stem cells. The results of our study provide new evidence that cancer stem cells may be originated by suppression of stem cell differentiation.


Subject(s)
Adipocytes/cytology , Cell Differentiation/drug effects , Mesenchymal Stem Cells/cytology , Methylcholanthrene/pharmacology , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Cell Transformation, Neoplastic , Cells, Cultured , Dexamethasone/pharmacology , Drug Combinations , Female , Indomethacin/pharmacology , Insulin/pharmacology , Mesenchymal Stem Cells/drug effects , Rats , Rats, Wistar
19.
Tumori ; 97(4): 540-6, 2011.
Article in English | MEDLINE | ID: mdl-21989446

ABSTRACT

AIMS: To investigate the expression of sodium/iodide symporter (NIS) and thyroid stimulating hormone receptor (TSHR) in human thyroid cancer. PATIENTS AND METHODS: NIS and TSHR mRNA levels quantified by real-time PCR as well as NIS and TSHR proteins evaluated by immunohistochemistry were examined in surgical specimens including 38 benign nodules, 32 thyroid carcinomas and 36 normal thyroid samples. RESULTS: NIS and TSHR mRNA levels in thyroid carcinomas were significantly lower than in benign nodules and normal thyroid samples (P <0.001). Interestingly, we found that NIS and TSHR mRNA expression in benign nodules had similar levels to those in normal thyroid tissues. However, NIS and TSHR protein expression in benign nodules and thyroid carcinomas was stronger than in normal thyroid samples (P <0.05) but mainly located in cytoplasm. In addition, there was a significant positive correlation between NIS and TSHR in benign nodules and normal thyroid samples (r = 0.551 and 0.667, respectively, P = 0.001 and 0.000, respectively) but there was no such correlation in thyroid carcinomas (r = 0.222, P = 0.376). CONCLUSIONS: In thyroid carcinomas, NIS and TSHR mRNA levels were lower but the proteins were overexpressed. The NIS protein mainly locates in the cytoplasm, which therefore lacks the ability of transporting and absorbing iodine in patients with thyroid carcinoma. In addition, there was no correlation between NIS and TSHR in thyroid cancer, which may explain why, even after TSH stimulation, 10-20% of these malignant tumors are unable to concentrate enough radioiodine for effective therapy.


Subject(s)
Goiter, Nodular/metabolism , Receptors, Thyrotropin/metabolism , Symporters/metabolism , Thyroid Gland/metabolism , Thyroid Neoplasms/metabolism , Adenocarcinoma, Follicular/metabolism , Adult , Aged , Carcinoma, Papillary/metabolism , China , DNA, Complementary/metabolism , Female , Gene Expression Regulation, Neoplastic , Humans , Immunohistochemistry , Male , Middle Aged , RNA, Messenger/metabolism , Real-Time Polymerase Chain Reaction , Receptors, Thyrotropin/genetics , Symporters/genetics , Up-Regulation
20.
Dose Response ; 9(3): 356-68, 2011.
Article in English | MEDLINE | ID: mdl-22013398

ABSTRACT

The aim of this study was to examine the early effects of low dose (12)C(6+) irradiation or X-ray on peripheral blood lymphocytes (PBL) of patients with alimentary tract cancer and to explore mechanisms that may be involved in an antitumor immune response. We found that the percentage of T lymphocyte subsets, the mRNA expression levels of IL-2 and IFN-γ in PBL, and their protein levels in supernatant were significantly increased 24 hours after exposure to low dose radiation. The effects were more pronounced in the group receiving 0.05Gy (12)C(6+) ion irradiation than the group receiving X-ray irradiation. There was no significant change in the percentage of NK cell subsets and TNF-α production of PBL. Our study suggests that low dose irradiation could alleviate immune suppression caused by tumor burden and that the effect was more pronounced for 0.05Gy high linear energy transfer (LET) (12)C(6+) irradiation.

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