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1.
Acta Pharmaceutica Sinica ; (12): 793-801, 2022.
Article in Chinese | WPRIM | ID: wpr-922901

ABSTRACT

Multicellular tumor spheroids (MCTS) can simulate the structure and metabolic characteristics of tumors in vivo, which is of great significance to study the metabolic phenotype of tumor cells and the mechanism of drug intervention. In this study, esophageal cancer MCTS were constructed, and MCTS frozen sections were prepared after treated with different formulations of paclitaxel (PTX) including common PTX injection, PTX liposome and albumin bound PTX. MCTS mass spectrometry imaging analysis method was established by using air flow assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI). The visualization of the permeation and enrichment process of PTX in MCTs after PTX treatment was realized, and the spatially resolved metabolomics of PTX injection group was studied. The results showed that the permeation and enrichment behavior of PTX in MCTs model were related to the formulations. The changes of endogenous metabolites in MCTs of esophageal cancer after treated with PTX injection had temporal and spatial characteristics. The metabolic changes of MCTS during the initial 0-4 hours were dominated by the down-regulation of middle-high polarity metabolites and some lipids in the central region of MCTS, while the metabolic changes of MCTS during 8-72 hours were mainly up-regulated by lipid metabolites in the peripheral region of MCTS. The combination of in vivo tumor-associated MCTs model with label free, highly sensitive and high coverage mass spectrometry imaging technology provided a new method and strategy for the study of pharmacometabolomics.

2.
International Eye Science ; (12): 2017-2021, 2019.
Article in Chinese | WPRIM | ID: wpr-756824

ABSTRACT

@#AIM: To investigate the protective effect of small interfering RNA targeting HIF-1α in retina of diabetic retinopathy mice and its mechanism. <p>METHODS: Totally 40 C57BL/6 male mice were randomly divided into the normal group, diabetes group, siRNA-HIF-1α group and siRNA-NC group. The diabetic models were constructed. The histopathological change of the retina of the mice was observed in each group. The microvessel density(MVD)was detected by immunohistochemistry. The expressions of HIF-1α, VEGF,NF-κB, IL-1, IL-6 and TNF-α mRNA in the retina in each group were detected by real-time quantitative PCR. The expressions of HIF-1α, ET-1 and vWF proteins in the retina in each group were detected by Western blot. <p>RESULTS: The body weights of diabetes group, siRNA-NC group and siRNA-HIF-1α group were lower than the normal group, while the blood glucose levels were higher than the normal group(All <i>P</i><0.05). The MVD in the diabetic group and siRNA-NC group were significantly higher than those in the normal group, while the siRNA-HIF-1α group were significantly lower than the diabetes group and siRNA-NC group(All <i>P</i><0.05). Compared with the diabetes group and siRNA-NC group, the relative expression levels of HIF-1α, VEGF, NF-κB, IL-1, IL-6 and TNF-α mRNA in the retina in the siRNA-HIF-1α group were decreased(All <i>P</i><0.05). Compared with the diabetes group and siRNA-NC group, the relative expression levels of HIF-1α,ET-1 and vWF proteins in the retina in the siRNA-HIF-1α group were decreased(All <i>P</i><0.05).<p>CONCLUSION: Specific silencing of HIF-1α gene could protect the retina of DR. The mechanism may be related to the reduction of angiogenesis and vascular endothelial injury.

3.
Article in Chinese | WPRIM | ID: wpr-692264

ABSTRACT

As a promising new molecular imaging technique,mass spectrometry imaging(MSI) has attracted more and more attention in the field of biomedicine. A method of air flow assisted ionization-ultra high resolution mass spectrometry-based mass spectrometric imaging (AFAI-MSI) was developed to profile endogenous metabolites in rat kidney tissue in this study. Rat kidneys were collected and cut into frozen tissue sections,and then were analyzed on an AFAI-MSI system in positive ion mode using acetonitrile-isopmpyl alcohol-water (4:4:2,V/V,5 μL/min) as spray solvent,N2as spray gas(0.6 MPa) and air as assisting gas (45 L/min). The mass range and resolution were set to be 70-1000 Da and 70000, respectively. As a result,a total of 38 metabolites, including organic amines, sugars, vitamins, peptides, neurotransmitters, organic acids,phospholipids,sphingolipids,glyceride,and cholesterol esters, were identified and imaged to characterize their tissue-specific distribution in kidney tissues, and some metabolites, such as choline, acetylcoline,betaine,phoshocholine,and glycerophosphocholine were found to have distinct distribution along the cortex-medulla axis,which may be involved in the formation of osmotic pressure gradient in the kidney. The proposed ultra high resolution mass spectrometry based AFAI-MSI method could work without sample pretreatment, showed high sensitivity and wide metabolite coverage, and was expected to provide a new analytical approach for the research of in situ characterization and metabolic regulation mechanism of endogenous metabolites in kidney.

4.
Acta Pharmaceutica Sinica ; (12): 1564-2016.
Article in Chinese | WPRIM | ID: wpr-779325

ABSTRACT

The research aimed to investigate the suppression effect of MaiShu which contains hawthorn, hippophae, medlar, phytosterols (β-sitosterol, stigmasterol and campesterol), β-glucan and lycopeneon formation of atherosclerotic plaque in apolipoprotein E knock-out (ApoE-/-) mice. Liquid chromatography-ultraviolet-mass spectrometry (LC-UV-MC) methods were used to analyze the main chemical composition of MaiShu.Atherosclerotic mice models were established by high-fat diet. The mice were administrated with MaiShu (1, 2, 4 g·kg-1·d-1) or other contrast materials by intragastric route for 10 weeks continuously. At the end of administration, the blood of mice was collected for tests of the serum total cholesterol (TC), total triglyceride (TG) and high density lipoprotein cholesterol (HDL-C) level. Atherosclerotic lesions in aorta and aortic root were assessed by calculating the relative area of lesions (oil red O stained). Intravital fluorescence microscopic system was used to evaluate the leukocyte-endothelial adhesion in mesenteric artery of mice by detecting the rolling velocity of white blood cells (WBC). Collagenous fibers and macrophages in lesions were detected by sirius red staining and immunological histological chemistry to evaluate the atherosclerotic plaque stability. Results showed that MaiShu contains various flavonoids (9.5%), phytosterols (23.8%) and polysaccharides (8.9%). The serum lipid level of model animals was significantly higher than the control animals. Serum TC level was decreased by MaiShu (4 g·kg-1, P-1) compared to model (P-1, P-1, P-1, P-1, P-1, P-1) and macrophages were decreased (2, 4 g·kg-1) compared to model. These results demonstrate that MaiShu can obviously decrease the serum lipid levels and the risk of leukocyte-endothelial adhesion in ApoE-/- mice. The effect of MaiShu may be associated with the decrease of macrophages in plaque.

5.
Acta Pharmaceutica Sinica ; (12): 444-2016.
Article in Chinese | WPRIM | ID: wpr-779190

ABSTRACT

The study aims to develop a rapid, specific and sensitive method for quantitative analysis of trace impurities in levofloxacin formulation using LC-MS/MS. The quality of the different formulations from 19 plants was evaluated in the contents of the impurities. The results indicated that there were 5 impurities in the samples, and the content was different in the products with same formulation by different plants. The products of 3 plants were in good quality with impurities level under 0.01%. Levofloxacin N4'-methyl quaternary impurity was first reported as the formulation impurity. The impurities were tightly correlated to the reservation of drug, process control of formulation and storage during transportation. The results suggest that our method is sensitive and specific to detect the trace impurities in formulation, and can be used to monitor the quality of commercial drug product.

6.
Article in Chinese | WPRIM | ID: wpr-330214

ABSTRACT

A new sesquiterpenoid, 8α-hydroxy-6β-methoxy-1-oxoeremophila-7 (11), 9 (10) -diene-12, 8-olide (1) and five known compounds, petasin (2), caffeic acid (3), hepta-cosanol (4), β-sitosterol (5) and β-daucosterol (6) have been isolated from the roots of Ligularia intermedia. The compounds were isolated by column chromatography on silica gel and Sephadex LH-20, and identified based on spectral analyses (MS, 1H-NMR, 13C-NMR).


Subject(s)
Asteraceae , Chemistry , Drugs, Chinese Herbal , Chemistry , Magnetic Resonance Imaging , Molecular Structure , Plant Roots , Chemistry , Spectrometry, Mass, Electrospray Ionization
7.
Acta Pharmaceutica Sinica ; (12): 1705-1709, 2013.
Article in Chinese | WPRIM | ID: wpr-298022

ABSTRACT

To screen the harmful substance 5-hydroxymethyl furfural content in commercially available traditional Chinese medicine injection which are commonly used, and to preliminarily evaluate the quality of these injections, 5-hydroxymethyl furfural was taken as an index. The contents of 5-hydroxymethyl furfural in 56 samples which consist of 23 kinds of traditional Chinese medicine injections and glucose injection were determined using LC-MS/MS, and 5-hydroxymethyl furfural was detected in 52 of these samples. The minimal content was 0.0038 microg x L(-1) and the maximum content was 1420 microg x mL(-1). The contents of 5-hydroxymethyl furfural were significantly different in traditional Chinese medicine injection which came from different kinds, manufacturers or batches. The results showed the quality difference of commercially available traditional Chinese medicine injection is significant taking 5-hydroxymethyl furfural content as assessment index. More attention should be paid to the safety of 5-hydroxymethyl furfural in traditional Chinese medicine injection, and unified limitation standard should be set to improve medication safety of traditional Chinese medicine injection.


Subject(s)
Chromatography, Liquid , Drug Combinations , Drugs, Chinese Herbal , Chemistry , Eleutherococcus , Chemistry , Furaldehyde , Glucose , Chemistry , Injections , Medicine, Chinese Traditional , Plant Extracts , Chemistry , Plants, Medicinal , Chemistry , Tandem Mass Spectrometry
8.
Chinese Journal of Cancer ; (12): 344-350, 2011.
Article in English | WPRIM | ID: wpr-294514

ABSTRACT

Recent findings show that Toll-like receptors (TLRs) expressed in immune cells play a crucial role in the innate immune response and the subsequent induction of adaptive immune responses against microbial infection on tissue injury. Furthermore, expression of TLRs in cancer cells is associated with tumor proliferation and invasion. To explore the role of TLRs expression in cervical carcinogenesis in Uighur women, we detected the expressions of TLR3, TLR4, TLR7, and TLR9 in 25 normal cervical tissues, 64 cervical intraepithelial neoplasia (CIN) tissues, and 63 cervical squamous cell carcinoma (CSCC) tissues using immunohistochemical staining, as well as human papillomavirus type 16 (HPV16) infection using PCR. All samples used in this study were from Xinjiang Uighur women. We found the expression levels of TLR4, TLR7, and TLR9 were significantly higher in CIN and CSCC than in normal controls (P < 0.05). Up-regulation of TLR4 and TLR7 were correlated with tumor differentiation but not FIGO stage or lymph node metastasis (P > 0.05). Up-regulation of TLR9 was correlated with lymph node metastasis (P < 0.05) but not tumor differentiation or FIGO stage (P > 0.05). We also analyzed the correlation between the expressions of TLRs and HPV16 infection and found that the expressions of TLR4 and TLR9 significantly correlated with HPV16 infection in CIN (r = 7.434, P = 0.006; r = 7.123, P = 0.008) and CSCC (r = 6.423, P = 0.001; r = 8.478, P = 0.004), whereas the expression of TLR3 was not significantly different in any of the three groups and had no significant correlation with HPV16 infection. Our results suggest that high expression of TLR4, TLR7, and TLR9 may play important roles in the development and progression of CIN and CSCC in Uighur women, and the expressions of TLR4 and TLR9 can be up-regulated by HPV16 infection.


Subject(s)
Carcinoma, Squamous Cell , Metabolism , Pathology , Virology , Cervical Intraepithelial Neoplasia , Metabolism , Pathology , Virology , China , Ethnology , Female , Gene Expression Regulation, Neoplastic , Human papillomavirus 16 , Humans , Lymphatic Metastasis , Neoplasm Staging , Papillomavirus Infections , Genetics , Pathology , Toll-Like Receptor 3 , Metabolism , Toll-Like Receptor 4 , Metabolism , Toll-Like Receptor 7 , Metabolism , Toll-Like Receptor 9 , Metabolism , Toll-Like Receptors , Metabolism , Up-Regulation , Uterine Cervical Neoplasms , Metabolism , Pathology , Virology
9.
Acta Physiologica Sinica ; (6): 547-554, 2010.
Article in Chinese | WPRIM | ID: wpr-337713

ABSTRACT

Abundant evidence has suggested that neuroinflammation participates in the pathogenesis of Parkinson's disease (PD). The emerging evidence has supported that microglia may play key roles in the progressive neurodegeneration in PD and might be a promising therapeutic target. Ganoderma lucidum (GL), a traditional Chinese medicinal herb, has been shown potential neuroprotective effect in our clinical trials that lead us to speculate that it might possess potent anti-inflammatory and immunomodulating properties. To test this hypothesis, the present study investigated the potential neuroprotective effect of GL and underlying mechanism through inhibiting microglial activation using co-cultures of dopaminergic neurons and microglia. The cultures of microglia or MES23.5 cells alone or together were treated for 24 h with lipopolysaccharide (LPS, 0.25 μg/mL) as a positive control, GL extracts (50-400 μg/mL) or MES23.5 cell membrane fragments (150 μg/mL) were used in treatment groups. Microglia activation, microglia-derived harmful factors and [(3)H]dopamine ([(3)H]DA) uptake of MES23.5 cells were analyzed. The results showed that microglia were activated by LPS and MPP(+)-treated MES23.5 cell membrane fragments, respectively. Meanwhile, GL extracts significantly prevented the production of microglia-derived proinflammatory and cytotoxic factors, including nitric oxide, tumor necrosis factor-α (TNF-α) and interleukin 1β (IL-1β), in a dose-dependent manner and down-regulated the TNF-α and IL-1β expressions on mRNA level. In addition, GL extracts antagonized the reduction of [(3)H]DA uptake induced by MPP(+) and microglial activation. In conclusion, these results suggest that GL may be a promising agent for the treatment of PD through anti-inflammation.


Subject(s)
Cell Line , Dopaminergic Neurons , Cell Biology , Down-Regulation , Humans , Interleukin-1beta , Metabolism , Materia Medica , Pharmacology , Microglia , Cell Biology , Metabolism , Neuroprotective Agents , Pharmacology , Nitric Oxide , Metabolism , Parkinson Disease , Reishi , Chemistry , Tumor Necrosis Factor-alpha , Metabolism
10.
Acta Pharmaceutica Sinica ; (12): 692-697, 2007.
Article in Chinese | WPRIM | ID: wpr-268594

ABSTRACT

In this review, we provide information on hydroxyl radical generation, trapping and detection methods, including electron spin resonance (ESR), electrochemistry detection (ECD), fluorescence detection, UV detection, chemoluminescence and mass spectrometry (MS). In addition, the advantages and disadvantages of the above methods were discussed.


Subject(s)
Chromatography, Liquid , Methods , Electron Spin Resonance Spectroscopy , Methods , Electrophoresis, Capillary , Methods , Hydroxyl Radical , Chemistry , Luminescent Measurements , Methods , Mass Spectrometry , Methods , Spectrophotometry, Ultraviolet , Methods , Spin Trapping , Methods
11.
Article in Chinese | WPRIM | ID: wpr-679648

ABSTRACT

Objective To investigate the correlation of spiral CT(SCT)features with pathology, microvessel density(MVD)、expression of vascular endothelial growth factor(VEGF)、matrix metalloproteinase-2(MMP-2)in colorectal carcinoma.Methods Forty patients with colorectal carcinoma confirmed by operation were examined by SCT.The resected tumor specimens were immunohistochemically stained for the expression of VEGF,MMP-2 and the calculation of MVD.Results The accuracy of SCT in depicting the pericolic and wall infiltration was 92.5%.The metastasis rates of colorectal cancer with pericolic infiltration and wall infiltration were 75.0% and 33.3%,respectively,the differences were statistically significant between the two groups(P

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