1.Up-regulation of the CXCR4 expression by hypoxia in maxillary sinus carcinoma cells IMC3.
Lizhong SU ; Zhiming ZHANG ; Wenyue JI
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2014;28(8):548-550
OBJECTIVE:
To investigate the expression of CXCR4 in maxillary sinus carcinoma cells IMC3 under hypoxia.
METHOD:
IMC3 cells were cultured for 6 h, 12 h, 24 h and 48 h under normoxia and hypoxia. Real-Time PCR was applied to detect the expression of mRNA of CXCR4 and immunohistochemisrty was applied to investigate its protein level.
RESULT:
CXCR4 mRNA level was about 0.035 under normal conditions, which was obviously upregulated by hypoxia. The mRNA levels after culturing under hypoxia for 6 h, 12 h, 24 h and 48 h were 0.283, 0.313, 0.426, 0.510 respectively. There was statistically significant difference between the mRNA levels of each two groups (P < 0.05, Mann-Whiney Test) with a time dependent course, except for the difference between the groups of 6 h and 12 h. Immunohistochemistry showed that there was almost negative staining for CXCR4 in the cell cultured in nomoxia, while stong positive staining of CXCR4 was observed in cells cultured in hypoxia . The positive staining was located mainly in the cell membrane and cytoplasm and little in the nucleus.
CONCLUSION
Hypoxia could induce expression of CXCR4 in IMC3 cells at both mRNA and ptrotein levels. The upregulation of CXCR4 by hypoxia showed an obvious time dependent course.
Cell Hypoxia
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Cell Line, Tumor
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Gene Expression Regulation, Neoplastic
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Humans
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Maxillary Sinus
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metabolism
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pathology
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Receptors, CXCR4
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metabolism
2.The effects of TiO_2 nano thin film on the migration and adherence of BLEC in vitro
Mei, SHAN ; Jingning WENG ; Wenyue, SU
Chinese Ophthalmic Research 2009;27(12):1073-1076
Objective The generate of posterior capsular opacification(PCO)is associated with the adhensive and migration of residual subcapsular lens epithelial cells(LECs).Titanium dioxide(TiO_2)nanometer is proved to have the ability of killing tumor cells and cultured bovine LECs.This study tried to observe the effects of TiO_2 nanometer thin film provoked by light on adhesiveness and migration of bovine LECs in vitro.MethodsThe fresh bovine lenses were obtained and cultured in DMEM containing 10% of newborn bovine serum.The second to fifth generation of cells were used in this experiment.The slide modified by TiO_2 photocatalyst film was prepared by sol-gel method.Cultured cells were seeded in filmed or unfilmed slides respectively and exposed to ultraviolet(wavelength 365 nm)for 20 or 40 minutes.The contact angle between water drop and slide was measured by droping method and the cells adhered to slides were calculated after 24 and 48 hours of culture.The growth status and migration distance of bovine LECs were assayed and compared between filmed and unfilmed groups.ResultsThe contact angle between water drop and slide was 0°±2°and 18.825°±2.342° in filmed and unfilmed group respectively,indicating a obviously smaller contact angle in TiO_2 filmed group than unfilmed one.The numbers of bovine LECs adhered to filmed slide was considerably reduced in TiO_2 filmed group compared with unfilmed group in different UVA exposure time(t_(0 min)=5.492,P=0.001;t_(20 min)=6.031,P=0.000;t_(40 min)=6.828,P=0.000).However,no significant difference was found in the numbers of adhensive cells among 3 UVA irradiation time points(F=1.278,P=0.297).The migration distance of the cells was significantly shorter in TiO_2 filmed group in comparison with unfilmed group in 24 and 48 hours after UVA irradiation(F_(group)=14.965,P=0.000;F_(time)=38.033,P=0.000).ConclusionThe TiO_2 nanometer thin film is characterized by the superhydrophilic property.So it can effectively impede the adhesion and migration of bovine LECs in vitro.
3.Rapid Quality Evaluation of Astragali Radix by AOTF-NIR
Xuhua REN ; Ting SU ; Wenyue JIANG ; Yadong LI ; Guangjing ZHENG ; Lu GAO
China Pharmacy 2018;29(2):168-171
OBJECTIVE:To establish the method for rapid quality evaluation of Astragali Radix.METHODS:The moisture of medicinal material was determined by oven drying method;the content of astragaloside Ⅳ was determined by HPLC-ELSD;the content of isoflavone glucoside was determined by HPLC (as reference value).The partial least squares (PLS) method combined with acousto-optic turnable filter-NIDRS was adopted to build quantitative model of above indexes in Astragali Radix (as predict value).According to reference value,60 batches of sample were collected.The spectra pretreatment was conducted by first derivative method combined with Savitzky golay.The optimal bands of moisture,astragaloside Ⅳ and isoflavone glucoside were 1 100-2 300 nm,1 080-2 160 nm,1 170-2 230 nm,respectively.RESULTS:The content determination of moisture,astragaloside Ⅳ and isoflavone glucoside in samples were all in line with methodology requirements.The corrected mean square root deviation of quantitative model for moisture,astragaloside Ⅳ,calycosin glucoside were 0.132 3,0.006 6,0.002 5,respectively;predicted mean square root deviation were 0.237 1,0.016 3,0.004 7;internal cross validation coefficient of correction set were 0.975 9,0.953 3,0.968 0;internal verification deviation of quantitative model were 1.43%,1.90%,1.84%;external verification deviation were 1.73 %,2.68 %,2.71%,respectively.CONCLUSIONS:The method is rapid,accurate,simple,pollution-free,and can be used for rapid quality evaluation of Astragali Radix.
4.Rapid Determination of Cryptotanshinone in Salvia miltiorrhiza by AOTF-NIDRS
Ting SU ; Wenyue JIANG ; Cheng XING ; Xuhua REN ; Xindong LI ; Xianli CUI ; Lu GAO
China Pharmacy 2018;29(8):1044-1048
OBJECTIVE:To establish rapid method for content determination of cryptotanshinone in Salvia miltiorrhiza. METHODS:The content of cryptotanshinone in sample was determined by HPLC(as reference value). AOTF-NIDRS combined with PLS was used to establish quantitative correction model for the content of cryptotanshinone in S. miltiorrhiza. According to the results of content determination of cryptotanshinone in samples,35 samples of medicinal material were collected. First-order derivative combined with smoothing filter coefficient method was used to pretreat spectrum,and optimal band range for content determination of cryptotanshinone in sample ranged 1 250-2 150 nm. RESULTS:Methodology validation of content determination of cryptotanshinone in sample was in line with the requirements. Correction mean square deviation of quantitative correction model of cryptotanshinone was 0.014 6,and predicted mean square deviation was 0.022 3,coefficient of association was 0.976 6. The internal verification deviation was 2.41% and the external verification deviation was 4.06%. CONCLUSIONS:This method is rapid,accurate,simple and pollution-free.It can be used for rapid content determination of cryptotanshinone in S.miltiorrhiza.
5.Rapid Judgement of Blending Endpoint of Jingqi Shuangshen Capsules and Content Determination of Astragaloside Ⅳ by AOTF-NIR
Ting SU ; Wenyue JIANG ; Yadong LI ; Xuhua REN ; Zijun MA ; Meixin QI ; Xianli CUI ; Lu GAO
China Pharmacy 2018;29(12):1616-1620
OBJECTIVE:To establish the method for rapid judgement of blending endpoint of Jingqi shuangshen capsules and content determination of astragaloside Ⅳ. METHODS:AOTF-NIR combined with principal component analysis and Moving Block Standard Deviation method was used to identify the blending endpoint. First derivative combined with savitzky-golay filter method were used to spectrum pretreatment. The partial least square method was used to establish quantitative analysis model of the content of astragaloside Ⅳin mixed endpoint sample. The content of astragaloside Ⅳ in mixed endpoint sample was determined by HPLC-ELSD to validate the model. RESULTS:Methodology validation of content determination of astragaloside Ⅳ in mixed material sample and mixed endpoint sample was in line with the requirements. NIR monitoring results showed that the product reached the blending endpoint after 30 min. The results of NIR monitoring were generally consistent with the results of HPLC-ELSD. The principal component dimension of the quantitative model was 9;determination coefficients was 0.954 9;Root Mean Square of Calibration of the model was 0.039 2;Root Mean Square Error of Prediction of the model was 0.042 6. Predicted average value of astragaloside Ⅳ by NIR was 11.74 mg/g,and measured average value of astragaloside Ⅳ by HPLC-ELSD was 11.38 mg/g;average deviation was 3.16%. CONCLUSIONS:AOTF-NIR can rapidly judge the blending endpoint sample of Jingqi shuangshen capsules,rapidly determine the content of astragalosideⅣin mixed endpoint material,improve the quality control level of blending process and shorten blending cycle.
6.Lyciumbarbarum polysaccharides ameliorate canine acute liver injury by reducing oxidative stress, protecting mitochondrial function, and regulating metabolic pathways.
Jianjia HUANG ; Yuman BAI ; Wenting XIE ; Rongmei WANG ; Wenyue QIU ; Shuilian ZHOU ; Zhaoxin TANG ; Jianzhao LIAO ; Rongsheng SU
Journal of Zhejiang University. Science. B 2023;24(2):157-171
The development of acute liver injury can result in liver cirrhosis, liver failure, and even liver cancer, yet there is currently no effective therapy for it. The purpose of this study was to investigate the protective effect and therapeutic mechanism of Lyciumbarbarum polysaccharides (LBPs) on acute liver injury induced by carbon tetrachloride (CCl4). To create a model of acute liver injury, experimental canines received an intraperitoneal injection of 1 mL/kg of CCl4 solution. The experimental canines in the therapy group were then fed LBPs (20 mg/kg). CCl4-induced liver structural damage, excessive fibrosis, and reduced mitochondrial density were all improved by LBPs, according to microstructure data. By suppressing Kelch-like epichlorohydrin (ECH)-associated protein 1 (Keap1), promoting the production of sequestosome 1 (SQSTM1)/p62, nuclear factor erythroid 2-related factor 2 (Nrf2), and phase II detoxification genes and proteins downstream of Nrf2, and restoring the activity of anti-oxidant enzymes like catalase (CAT), LBPs can restore and increase the antioxidant capacity of liver. To lessen mitochondrial damage, LBPs can also enhance mitochondrial respiration, raise tissue adenosine triphosphate (ATP) levels, and reactivate the respiratory chain complexes I‒V. According to serum metabolomics, the therapeutic impact of LBPs on acute liver damage is accomplished mostly by controlling the pathways to lipid metabolism. 9-Hydroxyoctadecadienoic acid (9-HODE), lysophosphatidylcholine (LysoPC/LPC), and phosphatidylethanolamine (PE) may be potential indicators of acute liver injury. This study confirmed that LBPs, an effective hepatoprotective drug, may cure acute liver injury by lowering oxidative stress, repairing mitochondrial damage, and regulating metabolic pathways.
Animals
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Dogs
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Antioxidants/metabolism*
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Carbon Tetrachloride
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Chemical and Drug Induced Liver Injury/drug therapy*
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Kelch-Like ECH-Associated Protein 1/metabolism*
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Liver
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Metabolic Networks and Pathways
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Mitochondria/metabolism*
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NF-E2-Related Factor 2/metabolism*
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Oxidative Stress
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Polysaccharides/pharmacology*
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Lycium/chemistry*