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Photocatalysts based on single atoms (SAs) modification can lead to unprecedented reactivity with recent advances. However, the deactivation of SAs-modified photocatalysts remains a critical challenge in the field of photocatalytic CO2 reduction. In this study, we unveil the detrimental effect of CO intermediates on Cu single atoms (Cu-SAs) during photocatalytic CO2 reduction, leading to clustering and deactivation on TiO2. To address this, we developed a novel Cu-SAs anchored on Au porous nanoparticles (CuAu-SAPNPs-TiO2) via a vectored etching approach. This system not only enhances CH4 production with a rate of 748.8â µmol â g-1 â h-1 and 93.1 % selectivity but also mitigates Cu-SAs clustering, maintaining stability over 7â days. This sustained high performance, despite the exceptionally high efficiency and selectivity in CH4 production, highlights the CuAu-SAPNPs-TiO2 overarching superior photocatalytic properties. Consequently, this work underscores the potential of tailored SAs-based systems for efficient and durable CO2 reduction by reshaping surface adsorption dynamics and optimizing the thermodynamic behavior of the SAs.
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It was to investigate the value of quantitative fluorescence PCR (QF-PCR) for the selection of specific short tandem repeat (STR) in prenatal diagnosis of fetal chromosomal diseases. Amniotic fluid (AF) and villus samples were obtained from 80 pregnant women at 16-20 weeks of gestation, and venous blood samples were obtained from 60 normal individuals to extract and prepare peripheral blood chromosome, AF cell chromosome, and villus cell chromosome samples for specific STR locus detection. It showed that the area ratio of AMX peak to AMY peak in the Genescan typing map of peripheral blood DNA of normal males was close to 1:1, while the Genescan typing map of peripheral blood DNA of normal females had only AMX peak and no AMY peak. Normal heterozygous individuals had an area ratio between 1 and 1.45 for venous blood, 1.002 and 1.27 for villous samples, and 1 and 1.35 for AF samples. The karyotype of a male fetus was 46, XY, inv [9] (p11: q13), and the structure of fetal chromosome 9 was inverted (interarm), and the site of structural inversion was band 1 in the short breech 1 region and band 3 in the long arm 1 region of chromosome 9. It suggested that QF-PCR can effectively identify the normal human body and cases by selecting specific STR locus detection, which has a good application value for prenatal diagnosis of fetal chromosomal diseases.
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Cromosomas Humanos Par 9 , Diagnóstico Prenatal , Embarazo , Humanos , Femenino , Masculino , Feto , Fluorescencia , Repeticiones de Microsatélite/genéticaRESUMEN
OBJECTIVE: To investigate the amino acid composition and content of common vegetables in Fujian Province, and to evaluate their nutritional value. METHODS: Selecting 25 kinds of common vegetables from Fujian, composition of amino acid in protein was determined according to GB/T 5009. 124-2016 national food safety standard-determination of amino acid. Evaluating method for nutritional value in food protein: to evaluate nutritional evaluation of vegetable protein by fuzzy recognition, essential amino acid index(EAAI), ratio coefficient(RC) and score ratio coefficient(SRC). RESULTS: Kinds of protein amino acid in vegetables were abundant, including 17 kinds of amino acids. Total amino acid content was from 224. 3 to 16202. 1 mg/100 g edible, the ratio of essential amino acids was from 0. 17 to 0. 44, RC was from 0. 13 to 1. 82, SRC was from 50. 56 to 82. 18, the first limiting amino acids of vegetable protein were methionine and cystine. CONCLUSION: There are various kinds of protein amino acids in vegetables, including all essential amino acids of human body. There is big difference among different kinds of amino acids.
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Aminoácidos , Verduras , Humanos , Metionina , Evaluación Nutricional , Valor NutritivoRESUMEN
Intensive insulin therapy has been extensively used to control blood glucose levels because of its ability to reduce the risk of chronic complications of diabetes. According to current guidelines, intensive glycemic control requires individualized glucose goals rather than as low as possible. During intensive therapy, rapid blood glucose reduction can aggravate microvascular and macrovascular complications, and prolonged overuse of insulin can lead to treatment-induced neuropathy and retinopathy, hypoglycemia, obesity, lipodystrophy, and insulin antibody syndrome. Therefore, we need to develop individualized hypoglycemic plans for patients with diabetes, including the time required for blood glucose normalization and the duration of intensive insulin therapy, which deserves further study.
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Skyrmions in two-dimensional (2D) magnetic materials are considered as ideal candidates for information carriers in next-generation spintronic devices. However, conventional methods for elucidating the physical properties of skyrmions have limited the development of skyrmions in diverse 2D magnetic material systems due to their requirements for electrical conductivity. To overcome this limitation, we propose to utilize an optical method (magneto-optical Kerr technique) to detect the skyrmions in 2D magnetic materials. Herein, the graphene/Fe3GeTe2/graphene vertical van der Waals (vdW) heterostructure devices are fabricated to generate stabilized skyrmions by applying out-of-plane current. In combination with magnetic circular dichroism measurements, we observe topological-reflective magnetic circular dichroism (T-RMCD) effects in Fe3GeTe2 flakes and attribute the peak-shaped component in T-RMCD to the annihilation of skyrmion magnetic domains. Notably, the T-RMCD signal can maintain up to a temperature as high as the Curie temperature of Fe3GeTe2 flakes (â¼200 K). Our work provides a universal, contactless, and nondestructive approach for studying the physical properties of skyrmions in 2D vdW magnetic materials while adding another degree of freedom to the modulation of skyrmions.
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This study presents a method based on acid transesterification and the purification by solid-phase extraction (SPE) coupled with gas chromatography-tandem mass spectrometry for quantifying 3- and 2-monochloropropanediol esters (3-MCPDE, 2-MCPDE) and glycidyl esters (GE) in nutritional foods. The fat was extracted by liquid-liquid extraction with petroleum ether and diethyl ether after the sample was hydrolysed with ammonia. Then the extract was purified by a SPE cartridge filled with the aminopropyl sorbents. It was demonstrated that the optimal elution volume for 3-MCPDE, 2-MCPDE and GE greatly depended on the sample matrix and varied from 6 to 12 mL for four different kinds of food matrices. All three analytes in the sample solution could be fully collected in the first 10-12 mL of eluate. By this way, monoacylglycerols commonly present in the samples were fully removed. Therefore, the overestimation of GE quantification was effectively eliminated. The modified analytical procedure was fully validated in a single laboratory and has been recommended as a Chinese Food Safety National Standard. In addition, two derivatisation agents, heptafluorobutyrylimidazole and phenylboronic acid, were proved to be equivalent in method accuracy and precision for the quantification of three analytes.
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Ésteres , Análisis de los Alimentos , Contaminación de Alimentos , Cromatografía de Gases y Espectrometría de Masas , Propanoles , Extracción en Fase Sólida , Espectrometría de Masas en Tándem , Ésteres/análisis , Hidrólisis , Contaminación de Alimentos/análisis , Propanoles/análisis , Propanoles/química , Compuestos Epoxi/análisis , Compuestos Epoxi/química , alfa-Clorhidrina/análisis , Ácidos/análisis , Ácidos/químicaRESUMEN
Background: The prognosis of patients who can achieve a complete response after neoadjuvant chemotherapy could be significantly improved. Thus, accurately predicting the efficacy of neoadjuvant chemotherapy is of great clinical significance. Currently, previous indicators such as neutrophil to lymphocyte ratio was poor in predicting the efficacy and prognosis of neoadjuvant chemotherapy in human epidermal growth factor receptor 2 (HER2) positive breast cancer patients. Methods: The data of 172 HER2 positive breast cancer patients admitted to the Nuclear 215 Hospital of Shaanxi Province from January 2015 to January 2017 were retrospectively collected. After neoadjuvant chemotherapy, the patients were divided into the complete response group (n=70) and the non-complete response group (n=102). The clinical characteristics and systemic immune-inflammation index (SII) levels of the two groups were compared. The patients were followed-up for 5 years post-surgery to observe whether recurrence or metastasis occurred after the operation by clinic visit combined with telephone calls. Results: The SII of the complete response group was significantly lower than that of the non-complete response group (587.43±175.97 vs. 821.82±231.58; P=0.000). The SII was valuable in predicting which HER2 positive breast cancer patients would fail to achieve a pathological complete response, and the area under the curve (AUC) was 0.773 [95% confidence interval (CI): 0.705-0.804; P=0.000]. A SII >755.10 was an adverse factor for HER2 positive breast cancer patients achieving a pathological complete response after neoadjuvant chemotherapy [P=0.000; relative risk (RR): 0.172 (95% CI: 0.082-0.358)]. The SII level was valuable in predicting recurrence within 5 years of surgery, and had an AUC of 0.828 (95% CI: 0.757-0.900; P=0.000). A SII >755.10 was a risk factor for recurrence within 5 years of surgery [P=0.001; RR: 4.945 (95% CI: 1.949-12.544)]. The SII level was valuable in predicting metastasis within 5 years of surgery, and had an AUC of 0.837 (95% CI: 0.756-0.917; P=0.000). A SII >755.10 was a risk factor for metastasis within 5 years of surgery [P=0.014, RR: 4.553 (95% CI: 1.362-15.220)]. Conclusions: The SII was associated with the prognosis and efficacy of neoadjuvant chemotherapy in HER2 positive breast cancer patients.
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Correction for 'General method for detecting acrylamide in foods and comprehensive survey of acrylamide in foods sold in Southeast China' by Li Yangping et al., Anal. Methods, 2023, 15, 2275-2283, https://doi.org/10.1039/D3AY00469D.
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BACKGROUND: CCL19 is a chemokine involved in cancer research due to its important role in the tumor microenvironment (TME) and clinical relevance in cancers. This study aimed to analyze transcription expression, genomic alteration, association with tumor immune microenvironment of CCL19 expression and its prediction value for prognosis and responses to immunotherapy for patients with cancers. METHODS: RNA sequencing data and corresponding clinicopathological information of a total of large-scale cancer patients were obtained from The Cancer Genome Atlas and Gene Expression Omnibus databases. Multiplex immunofluorescence (mIF) was implemented to identify differential infiltration of Treg, CD8+ T cells, and tumor-associated macrophages, while CCL19 immunohistochemistry was conducted on 182 breast cancer samples from a real-world cohort. RESULTS: Based on large-scale multi-center survival analysis of cancer patients, we found the prognosis of patients with high CCL19 expression was prominently better than those with low CCL19 expression. For patients from multiple independent cohorts, suppressed CCL19 expression exerts significant progressive phenotype and apoptosis activity of cancers, especially in breast and ovarian cancer. Interestingly, anti-tumor immune cells, specifically the CD8+ T cells and macrophages, were clustered from TME by elevated CCL19 expression. Additionally, higher CCL19 levels reflected heightened immune activity and substantial heterogeneity. CONCLUSIONS: In conclusion, our findings support the notion that elevated CCL19 expression is linked to favorable outcomes and enhanced anti-tumor immunity, characterized by increased CD8+ T cells within the TME. This suggests the potential of CCL19 as a prognostic marker, predictive biomarker for immunotherapy, therapeutic target of cancers.
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Linfocitos T CD8-positivos , Neoplasias Ováricas , Humanos , Femenino , Pronóstico , Microambiente Tumoral , Quimiocinas , Quimiocina CCL19RESUMEN
2D ferromagnet is a good platform to investigate topological effects and spintronic devices owing to its rich spin structures and excellent external-field tunability. The appearance of the topological Hall Effect (THE) is often regarded as an important sign of the generation of chiral spin textures, like magnetic vortexes or skyrmions. Here, interface engineering and an in-plane current are used to modulate the magnetic properties of the nearly room-temperature 2D ferromagnet Fe5 GeTe2 . An artificial topology phenomenon is observed in the Fe5 GeTe2 /MnPS3 heterostructure by using both anomalous Hall Effect and reflective magnetic circular dichroism (RMCD) measurements. Through tuning the applied current and the RMCD laser wavelength, the amplitude of the humps and dips observed in the hysteresis loops can be modulated accordingly. Magnetic field-dependent hysteresis loops demonstrate that the observed artificial topological phenomena are induced by the generation and annihilation of the magnetic domains. This work provides an optical method for investigating the topological-like effects in magnetic structures and proposes an effective way to modulate the magnetic properties of magnetic materials, which is important for developing magnetic and spintronic devices in van der Waals magnetic materials.
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Cervical cancer is a frequently reported cancer of reproductive tract in females and is worldwide 4th most common malignant tumor. The present study investigated the effect of vanillin oxime on proliferation of cervical cancer cells. Vanillin oxime treatment led to suppression of Caski cell proliferation but could not affect proliferation of (HCvEpC) cells at the tested (2 to 10 µM) concentrations. In vanillin oxime treated Caski cells ROS level showed an increase with enhancement in concentration from 2 to 10 µM. Vanillin oxime treatment significantly (P<0.0487) lowered the count of colonies and inhibited invasive abilities of Caski cells. Treatment with vanillin oxime caused a significant (P<0.0487) suppression in HIF1α expression in Caski cells. Caski cell apoptotic count reached to 8.76% and 48.65%, on incubation with 2 and 10 µM concentrations of vanillin oxime respectively. After treatment with vanillin oxime a prominent reduction in MMP-2 and -9 levels was observed in Caski cells. A prominent reduction in p-ERK1/2 and p-Akt levels was observed in Caski cells after treatment with vanillin oxime. Vanillin oxime inhibits cervical cancer proliferation, invasive abilities, induces apoptotic signalling, and elevates ROS production. Therefore, vanillin oxime may be developed as an effective therapeutic agent for treatment of cervical cancer.
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Two-dimensional (2D) magnets are crucial in the construction of 2D magnetic and spintronic devices. Many devices, including spin valves and multiple tunneling junctions, have been developed by vertically stacking 2D magnets with other functional blocks. However, owing to limited local interactions at the interfaces, the device structures are typically extremely complex. To solve this problem, the nonlocal manipulation of magnetism may be a good solution. In this study, we use the magneto-optical Kerr effect technique to demonstrate the nonlocal manipulation of magnetism in an itinerant 2D ferromagnet, Fe3GeTe2 (FGT), whose magnetism can be manipulated via an antiferromagnet/ferromagnet interface or a current-induced spin-orbital torque placed distant from the local site. It is discovered that the coupling of a small piece of MnPS3 (â¼40 µm2) with FGT can significantly enhance the coercive field and emergence of exchange bias in the entire FGT flake (â¼2000 µm2). Moreover, FGT flakes with different thicknesses have the same coercive field at low temperatures if they are coupled together. Our study provides an understanding of the basic magnetism of 2D itinerant ferromagnets as well as opportunities for engineering magnetism with an additional degree of freedom.
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Fe3GeTe2/MnPS3 and Fe3GeTe2/MnPSe3 van der Waals heterostructures were fabricated by mechanical exfoliation. Via the magneto-optical Kerr effect and reflected magnetic circular dichroism measurements, we have observed nearly three times enhancement of the coercive field, improvement of Curie temperature, and exchange bias effect in both heterostructures. These observations may provide new insights into the emergent heterostructure devices between itinerant ferromagnets and metal thio- and selenophosphates for both applied and fundamental research studies in magnetic correlations.
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Increasing evidence indicates that DNA polymerase epsilon (POLE), which mediates DNA damage repair, is significantly associated with tumor prognosis. This study aimed to analyze POLE expression in tumor samples and its prognostic value for patients with clear cell renal cell carcinoma (ccRCC). We found significantly elevated POLE expression in ccRCC tissues compared with normal tissues of multiple independent cohorts. The POLE expression levels of 523 patients with ccRCC (The Cancer Genome Atlas RNA-seq data) and 179 patients with ccRCC with immunohistochemical data (Fudan University Shanghai Cancer Center) were analyzed to investigate the prognostic implications of POLE expression. Cox regression analyses were implemented to explore the effect of POLE expression on the prognosis of pan-cancer. These findings revealed that elevated POLE expression levels significantly correlated with shorter overall survival (p < 0.001, n = 701) of patients with ccRCC. These data indicate that POLE expression may serve as a prognostic biomarker for cancers. Although POLE mutations were not significantly associated with survival benefits conferred upon patients with ccRCC, a CD4+ T cell-regulated immune microenvironment was significantly activated. Moreover, we found that POLE expression in cancers significantly correlated with an immunosuppressive tumor microenvironment, higher intratumoral heterogeneity, and expression of immune checkpoint genes PDCD1, CTLA4, and CD86, possibly mediated via the JAK/STAT and Notch signaling pathways. In conclusion, the present study is the first to our knowledge to indicate that elevated POLE expression is significantly associated with poor survival and an immune-suppressive tumor microenvironment in ccRCC. These findings suggest that POLE can serve as a biomarker for guiding molecular diagnosis and facilitating the development of novel individual therapeutic strategies for patients with advanced ccRCC.
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In our program to synthesize a series of novel derivatives as potential analogs of honokiol for anti-tumor treatment, we have found that at least three of the derivatives of honokiol showed more potency to inhibit the proliferation of K562 leukemia cells and SPC-A1 adenocarcinoma cells. As a critical step to our further series synthesis of derivatives of honokiol, three derivatives of honokiol composed of two isomers and one compound with two formyl groups, which were hardly separated by common purification methods, needed to be rapidly separated and purified. The present work describes analytical and preparative high-speed counter-current chromatography (HSCCC) for the isolation and purification of these three C-formylation derivatives of honokiol, named 3'-formylhonokiol, 5-formylhonokiol and 3',5-diformylhonokiol, respectively. The solvent system for HSCCC separation was composed of hexane-ethyl acetate-methanol-water with the ratio of 1:0.4:1:0.4 (v/v). The one-step purification produced 157.8 mg, 121.6 mg and 21.2 mg of 3'-formylhonokiol, 5-formylhonokiol, 3',5-diformylhonokiol from crude sample of 400mg with purities of 98.6%, 99.2% and 99.6%, respectively, in an elution time of 2.5 h. The purities and structural identification were determined by HPLC, (1)H NMR, (13)C NMR and mass spectroscopy. Their anti-proliferation effects on K562, A549 and SPC-A1 cell lines were evaluated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay.
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Compuestos de Bifenilo/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Distribución en Contracorriente , Lignanos/aislamiento & purificación , Compuestos de Bifenilo/análisis , Compuestos de Bifenilo/química , Lignanos/análisis , Lignanos/química , Espectroscopía de Resonancia Magnética , Espectrometría de MasasRESUMEN
OBJECTIVES: To examine the efficacy and safety ofpersonalized tongxie formulas; to decrease type II errors to minimum. METHODS: Patients were randomized (1:1:1) into three groups given tongxie, placebo, or pinaverium 3 times daily for 4 weeks. Patients in the tongxie group were treated with personalized formulas based on TCM differential diagnosis, i.e., basic type of IBS, IBS due to liver depression and qi stagnation, excess heat in the liver, deficient spleen function, deficient kidney function, and others (groups 1-6). Primary endpoints were significantly greater reductions in abdominal pain and Bristol stool score. Secondary endpoints were reductions in pain and stool frequencies and abdominal discomfort and its frequency. RESULTS: There were significantly more patients whose stool consistencies were improved than pains were relieved in the entire population (p < 0.001), but there was no significantly difference in subpopulation group 3 (p > 0.05). There were significantly more patients whose stool frequencies were reduced than pain frequencies were reduced in the entire population (p < 0.001), but there were no significantly difference in the subpopulation Groups 1, 3, 4, and 6 (p > 0.05). Multiple active ingredients and their mechanisms of actions to relieve IBS symptoms were identified. CONCLUSION: The outcomes in subpopulations may be different from those of the entire population, indicating that personalized formulas are important to achieve optimal outcomes; the active ingredients and innovative mechanisms identified in this study can be the candidates for developing new IBS drugs, and used to manage IBS, respectively. TRIAL REGISTRATION: NCT01641224 (www.ClinicalTrials.gov).
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Medicamentos Herbarios Chinos/uso terapéutico , Síndrome del Colon Irritable/tratamiento farmacológico , Síndrome del Colon Irritable/epidemiología , Medicina de Precisión/estadística & datos numéricos , Adulto , Diagnóstico Diferencial , Femenino , Humanos , Síndrome del Colon Irritable/diagnóstico , Síndrome del Colon Irritable/fisiopatología , Masculino , Persona de Mediana Edad , Resultado del Tratamiento , Adulto JovenRESUMEN
Inhalational anthrax is a serious biothreat. Effective antibiotic treatment of inhalational anthrax requires early diagnosis; the further the disease has progressed, the less the likelihood for cure. Current means for diagnosis such as blood culture require several days to a result and require advanced laboratory infrastructure. An alternative approach to diagnosis is detection of a Bacillus anthracis antigen that is shed into blood and can be detected by rapid immunoassay. The goal of the study was to evaluate detection of poly-γ-D-glutamic acid (PGA), the capsular antigen of B. anthracis, as a biomarker surrogate for blood culture in a rabbit model of inhalational anthrax. The mean time to a positive blood culture was 26 ± 5.7 h (mean ± standard deviation), whereas the mean time to a positive ELISA was 22 ± 4.2 h; P = 0.005 in comparison with blood culture. A lateral flow immunoassay was constructed for detection of PGA in plasma at concentrations of less than 1 ng PGA/ml. Use of the lateral flow immunoassay for detection of PGA in the rabbit model found that antigen was detected somewhat earlier than the earliest time point at which the blood culture became positive. The low cost, ease of use, and rapid time to result of the lateral flow immunoassay format make an immunoassay for PGA a viable surrogate for blood culture for detection of infection in individuals who have a likelihood of exposure to B. anthracis.
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Carbunco/diagnóstico , Anticuerpos Antibacterianos/inmunología , Anticuerpos Monoclonales/inmunología , Antígenos Bacterianos/inmunología , Cápsulas Bacterianas/inmunología , Ácido Poliglutámico/análogos & derivados , Infecciones del Sistema Respiratorio/diagnóstico , Animales , Carbunco/microbiología , Afinidad de Anticuerpos/inmunología , Bacillus anthracis/inmunología , Biomarcadores , Diagnóstico Precoz , Inmunoensayo/métodos , Inmunoglobulina G/inmunología , Pruebas Inmunológicas/métodos , Ácido Poliglutámico/sangre , Ácido Poliglutámico/inmunología , Conejos , Infecciones del Sistema Respiratorio/microbiologíaRESUMEN
The rapid diagnosis of septicaemic melioidosis will have an impact on reduction of mortality. Currently, this relies almost exclusively upon culture of the causative agent Burkholderia pseudomallei from clinical samples. In acute sepsis, blood is the preferred specimen for culture and therefore should be the target for a rapid diagnostic tool. A lateral flow immunoassay (LFI) for the detection of B. pseudomallei antigen has been developed. This was compared with molecular detection using the targets T3SS1 and IpxO. Forty-five clinical samples of EDTA blood, which were culture-positive, were tested using both modalities. The LFI had a sensitivity of 40 %, whilst molecular detection had a sensitivity of 20 %. The poor performance of molecular detection has been described previously and is largely related to the use of whole-blood specimens collected into blood tubes containing EDTA. Whilst suboptimal, the LFI would be an adjunct in the rapid diagnosis of melioidosis.
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Antígenos Bacterianos/análisis , Sangre/microbiología , Burkholderia pseudomallei/aislamiento & purificación , Cromatografía de Afinidad/métodos , Melioidosis/diagnóstico , Antígenos Bacterianos/inmunología , Burkholderia pseudomallei/inmunología , Humanos , Técnicas de Diagnóstico Molecular/métodos , Sensibilidad y Especificidad , Factores de TiempoRESUMEN
BACKGROUND: According to the traditional view, we depend on three methods to treat tumors; surgery, chemotherapy and radiotherapy. However, these methods have its own limitations in application. Traditional Chinese Medicine (TCM) is one of the oldest healing systems. Astragalus mongholicus (AMs) that is the common herbal medicine, the biggest part of TCM, have been proved to be effective in treating cancers from lots of clinical cases. However, we have not fully understood the anti-tumor mechanism of AMs, and this has lead to some doubt for some Western-Medicine scholars and restricts its wide use. The main objective of this research is to discuss the effect and mechanism of AMs to human stomach cancer. MATERIALS AND METHODS: To observe the effect and mechanism of tumor treatment by AMs, we have done the research from three major aspects, the influence of DCs, the inhibition of tumor in vitro as well as the animal studies in vivo after treatment. First, we culture the mouse dendritic cells (DCs) from bone marrow of mouse hind legs according to the method using Interleukin-4(IL-4) and Granulocyte-macrophage colony stimulating factor (GM-CSF), which refer to the way established by Inaba (Inaba K, 1992). And then we investigate the growth-rate of the DCs co-cultured with AMs injection. We analyze the expression of the Toll-like-receptor 4 (TLR4), with SYBR-Green I Real-time PCR and the I-kappa-B-alpha (IκB-α) with Western-Blot, the main regulatory protein to control nuclear factor NFκB-p65 nuclear translocation. Second, we choose the human gastric cancer cell lines MKN 45 as the target cell, which was co-cultured with DCs, T cells from spleen of mouse and AMs injection, and use MTT assay to judge the amount of cell lines and Immnunoflurescene to analyze the expression of anti-active caspase 3 pAb anti-PARP P85 fragment pAb, the mark of apoptosis of cells. Third, we have conducted the animal studies beside the basic experiment in vitro. The nude mouse developed stomach cancer, due to intra-preritoneal injection with MKN45 have been divided into two groups: the treatment group challenged with AMs injection and the control group with saline injection. We took the average of the diameter of each group as the y axis and the days after administered with AMs as x axis. After 40 days, all animals were killed by detruncation, and the tumor were removed and measured. We compare the diameter (<40 days) and weight (>40 days) of the tumor as well as the survival days between different groups to investigate the effect of inhibition of cancer. RESULTS: All results show that AMs is effective in treating human stomach cancer and the mechanism might be regulated by TLR4 mediated signal transduction of DCs. The results are briefly introduced as follows: First, we succeed in culturing the DCs induced by IL-4 and GM-CSF and find the positive rate of CD11c expression, the mark of DCs, is beyond 90% (Fig-1). We detect AMs can precipitate DCs maturation by upregulating TLR4 in SYBR-Green I Real-time PCR (Fig-2) and suppressing I.B-aby Western-Blot (Fig-3). Second, after the MKN45 co-cultured with DCs, T cells and AMs injection, the result show that AMs can great reduce the amount of cell lines by MTT assay (Fig-4) and induce apoptosis with Immunofluorescence (Fig-5). Finally, we have conducted animal studies beside the experiment in vitro, and the result in vivo show that AMs can delay tumor development from the diameter and weight of the tumor (Fig-6, Fig-7), prolong life-span and improve life-quality. Figure 1the morphology and phenotypic identification of DCs.The form of DCs observed by microscope with field 20*.The isotype antibody control using FCM.The positive rate of CD11c expression.Figure 2the melting curve and the chart of TLR4 expressiona) the melting curve of beta-actin; b)the melting curve of TLR4;c)the TLR4 expression of DCs stimulated with AM at different dose. There is significant statistic difference between the 60ng/mL and 80ng/mL group and other group (P<0.05 by rank test)Figure 3the IκB-α expression of DCs with different dose of AMsL0: 0ng/mL; L1:20ng/mL; L2:40ng/mL; L3:60ng/mL; L4:80ng/mLFigure 4MTT assay to analyse the viability and proliferation of the two cell lines (P<0.05 between the group with the dose of 60ng/mL and 80ng/mL and other group). the horizontal axis is the group treated with AM and saline at different dose, the vertical axis is the cell number.Figure 5the anti-active caspase-3 pAb (a) and anti-PARP P85 fragment pAb (b) actived by immunofluorescence.The cell mix were treated with 100uL anti-active caspase-3 pAb at a 1:250 dilution and anti-PARP P85 fragment pAb at a 1:100 dilution, and the secondary Ab was donkey anti-rabbit Cy®3 conjugate diluted 1:500 in PBS (Jackson Cat#711-165-152). From the photo, we find that anti-active caspase-3 pAb and anti-PARP P86 fragment pAb can express which is very important to indicate cell-apoptosis.Figure 6The difference of tumor between treatment group and control group. CONCLUSION: Ams Can play a great role in treating human stomach cancers as a good Chinese herbal medicine by precipitating DCs maturation, which is probably due to its effects by regulating the TLR4 mediated signal transduction.
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Planta del Astrágalo/química , Células Dendríticas/efectos de los fármacos , Medicamentos Herbarios Chinos/administración & dosificación , Transducción de Señal/efectos de los fármacos , Neoplasias Gástricas/tratamiento farmacológico , Receptor Toll-Like 4/metabolismo , Animales , Apoptosis/efectos de los fármacos , Células Dendríticas/metabolismo , Femenino , Humanos , Interleucina-4/genética , Interleucina-4/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Desnudos , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Receptor Toll-Like 4/genéticaRESUMEN
Burkholderia pseudomallei is a soil-dwelling bacterium and the causative agent of melioidosis. Isolation of B. pseudomallei from clinical samples is the "gold standard" for the diagnosis of melioidosis; results can take 3-7 days to produce. Alternatively, antibody-based tests have low specificity due to a high percentage of seropositive individuals in endemic areas. There is a clear need to develop a rapid point-of-care antigen detection assay for the diagnosis of melioidosis. Previously, we employed In vivo Microbial Antigen Discovery (InMAD) to identify potential B. pseudomallei diagnostic biomarkers. The B. pseudomallei capsular polysaccharide (CPS) and numerous protein antigens were identified as potential candidates. Here, we describe the development of a diagnostic immunoassay based on the detection of CPS. Following production of a CPS-specific monoclonal antibody (mAb), an antigen-capture immunoassay was developed to determine the concentration of CPS within a panel of melioidosis patient serum and urine samples. The same mAb was used to produce a prototype Active Melioidosis Detect Lateral Flow Immunoassay (AMD LFI); the limit of detection of the LFI for CPS is comparable to the antigen-capture immunoassay (â¼0.2 ng/ml). The analytical reactivity (inclusivity) of the AMD LFI was 98.7% (76/77) when tested against a large panel of B. pseudomallei isolates. Analytical specificity (cross-reactivity) testing determined that 97.2% of B. pseudomallei near neighbor species (35/36) were not reactive. The non-reactive B. pseudomallei strain and the reactive near neighbor strain can be explained through genetic sequence analysis. Importantly, we show the AMD LFI is capable of detecting CPS in a variety of patient samples. The LFI is currently being evaluated in Thailand and Australia; the focus is to optimize and validate testing procedures on melioidosis patient samples prior to initiation of a large, multisite pre-clinical evaluation.