1.Electrochemiluminescence Biosensor Based on DNAzyme and 3,4,9,10-Perylenetetracarboxylic Dianhydride Derivative Functionalized Hollow Gold Nanoparticles for Detection of Lead Ions
Xue LI ; Anyi CHEN ; Ying ZHUO ; Ruo YUAN
Chinese Journal of Analytical Chemistry 2015;(11):1701-1707
Based on target cycling amplification and 3 ,4 ,9 ,10-perylenetetracarboxylic dianhydride derivative functionalized singal probe, an ultrasensitive electrochemiluminescence ( ECL) sensor was designed for the detection of lead ions. The hairpin substrate DNA was immobilized on the electrode through molecular self-assembly. In the presence of Pb2+and DNAzyme, the substrate was cleaved with single strand DNA fragments left on the electrode surface. Meanwhile, the target and DNAzyme was released for another cleaving circularly. As a result, the single strand DNA fragments hybridized with the assist hairpin probe H1, which leaded to the fabrication of H2 labeled with the 3 , 4 , 9 , 10-perylenetetracarboxylic dianhydride derivative functionalized hollow gold nanoparticles. With the increasing concentration of Pb2+, much more signal probe was been captured and the ECL signal of the biosensor in peroxydisulfate ( S2 O2-8 ) solution would increase. An ECL assay demonstrates that the sensor has a good linear response to Pb2+ concentration in the range of 1í10-12 mol/L-1í10-6 mol/L, with a detection limit of 1í10-12 mol/L. The fabricated sensor shows good selectivity toward Pb2+against other common metal ions.
2.Isolation and purification of pig islet cells
Anyi QIAO ; Wenhong ZHANG ; Xinjie CHEN ; Shounan YI ; Yide OIAN ; Guohong XIAO ; Xuehu XU ; Yangxi HU
International Journal of Surgery 2009;36(5):297-299,封3
Objective To establish effective method for large-scale purification of islet cells from pig pan-cress. Methods Pig pancreas tissue was digested with collagenase P followed by purification in a HCA-Fi-coil dis continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity were evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release as-say in vitro. Results The number of the islets coll ected from each pancreas averaged (275 000±20 895)islet equivalents (IEQ) before purification, and (230 350±26 679) IEQ after the purification with discon-tinuous gradient centrifugation. From each gram of the pancreatic tissue, (2710±229) IEQ were obtained with an average purity of (50.2±1.95) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 4. 74-fold increase of insulin secretion over the basal level (3.3 mmol/L, P <0.001). Conclusion The established method can be applicable for large-scale purifi-cation of fully functional islet cells from pig pancreas.
3.Effects of N,N'-dinitrosopiperazine on cell proliferation in nasal and nasopharyngeal epithelia and expression levels of TRAF2 and p16 genes in transgenic mice
Yingchun HE ; Daofa TIAN ; Fangguo LU ; Jieqiong JIANG ; Anyi HE ; Yan CHEN
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(09):-
OBJECTIVE To investigate the effects of N,N'-Dinitrosopiperazine (DNP) on the cell proliferation of nasal and nasopharyngeal epithelia and the expression levels of TRAF2 and p16 genes in TgN (p53mt-LMP1)/HT transgenic mice and the relationships between them.METHODS The epithelial proliferating characteristics of nasal cavity and nasopharynx in TgN (p53mt-LMP1)/HT cancerous lesion inducing group treated by DNP (TI),TgN (p53mt-LMP1)/HT controlling group (TC), C57BL/6J cancerous lesion inducing group (CI) treated by DNP and C57BL/6J controlling group (CC) were observed for pathological evaluation by HE staining,and the expression levels of TRAF2 and p16 genes in these tissue samples were determined by immunohistochemical methods.RESULTS The occurring rates of precancerous lesions in nasal and/or nasopharyngeal epithelia in TI,TC,CI and CC groups were 90%,10%,0 and 0,respectively (P
4.Expression of CD24 in prostate cancer tissues and its clinical significance
CHEN Qing ; HUANG Zhiyang ; ZHANG Wenbin ; ZHENG Zhixiong ; SUN Anyi
Chinese Journal of Cancer Biotherapy 2019;26(12):1383-1386
Objective: To investigate the expression of CD24 in prostate cancer (PC) tissues, and explore its relationship with clinicopathological features of PC patients. Methods:Atotal of 40 cases of PC tissues and 36 cases of corresponding para-cacerous tissues resected during surgery at the Department of Urology Surgery, Quanzhou First HospitalAffiliated to Fujian Medical University from February 2016 to March 2019 were collected for this study; in addition, 46 cases of benign prostatic hyperplasia tissues were collected from patients underwent TURP surgery. Flow cytometry was used to detect the expression of CD24 in above mentioned tissues; One-way analysis of variance was used to analyze the relationship between the expression of CD24 and the age, tumor distribution, preoperative serum PSA, postoperative Gleason score, clinical stage and distant metastasis of PC patients. Results: The positive expression rate and MFI (mean fluorensece intensity) value of CD24 in prostate cancer tissues were significantly higher than those in para-cancerous prostate tissues and benign prostatic hyperplasia tissues (all P<0.05); CD24 positive expression rate and MFI value in PC tissues of patients with preoperative serum PSA≥10 ng/ml, postoperative Gleason score≥8 (low differentiation), clinical stage of T4 and distant metastasis were significantly higher than corresponding control group (all P<0.05); The expression of CD24 gradually increased with the progression of postoperative Gleason score and clinical stage ( P <0.05). Conclusions: The expression of CD24 is increased in prostate cancer tissues. The detection of CD24 expression level can help to determine the occurrence, development, invasion and metastasis of prostate cancer, and has potential clinical application value.
5.Efficacy and safety of intra-arterial thrombolysis for retinal artery occlusion based on the green channel for eye stroke
Anyi LIANG ; Yongyi NIU ; Qiaowei WU ; Daiyu CHEN ; Hongbin LIN ; Mingkui TAN ; Yijun HU ; Honghua YU
Chinese Journal of Ocular Fundus Diseases 2023;39(6):444-450
Objective:To explore the short-term efficacy and safety of intra-arterial thrombolysis (IAT) in the treatment of retinal artery occlusion (RAO) with the assistance of the rescue green channel in the eye stroke center.Methods:A prospective, interventional, single-center study. Thirty-eight eyes from 38 RAO patients who received IAT treatment in Guangdong Provincial People’s Hospital were enrolled. All the patients were rescued via the green channel in our eye stroke center. Data from comprehensive ocular examinations including best-corrected visual acuity (BCVA) and optical coherence tomography angiography (OCTA) were collected. BCVA was measured with Snellen chart and converted to the logarithmic minimum angle of resolution (logMAR) unit for statistical analysis. RTVue XR OCTA was used to measure vascular densities (VD) of the superficial capillary plexus (SCP), deep capillary plexus (DCP) and radial peripapillary capillary (RPC), and central retinal thickness (CRT). All RAO eyes attempted IAT treatment and 34 eyes were successful. Four eyes failed to complete IAT because of the occlusion of internal or common carotid arteries on the same side with the RAO eyes. Ocular examinations in post-operative 1-3 days were performed with the same devices and methods as those before surgery. Parameters measured before and after surgery include BCVA, VD of SCP, DCP, RPC, and CRT. Data of the green channel collected include the time intervals from onset of RAO to first presentation in local hospitals, and from onset of RAO to our eye stroke center. Comparisons of VD and CRT between the RAO eyes and contralateral healthy eyes were performed with independent samples Mann-Whitney U test; comparisons of VD and CRT in RAO eyes before and after IAT surgery were performed with paired samples Wilcoxon Rank Sum test. Results:Among the 34 RAO patients who had successful IAT surgery, 18 (52.9%, 18/34) were males and 16 (47.1%, 16/34) were females; the mean age was (51.0±12.9) years old. There were 30 and 4 eyes diagnosed as central RAO and branch RAO respectively. The logMAR BCVA before and after IAT surgery was 2.52±0.61 and 2.18±0.85 respectively, and the difference was statistically significant ( Z=-3.453, P=0.002). Before surgery, VD of SCP, DCP and RPC were significantly decreased and CRT was significantly increased in the affected eye compared with the contralateral healthy eyes, with the statistical significance ( P<0.001). Compared with those before surgery, the VD of SCP and DCP were significantly improved after surgery ( Z=-2.523, -2.427; P=0.010, 0.014), while there was no difference in VD of RPC and CRT ( Z=-1.448, -1.454; P=0.150, 0.159). The time interval between onset of RAO and first visit to the hospital was (6.56±6.73) hours; the time interval between onset of RAO and the arrival at our hospital was (24.11±19.90) hours. No cerebral stroke was observed in the early postoperative period and no cerebrocardiovascular events were observed later. he time interval between onset of RAO and the arrival at our hospital was (24.11±19.90) hours. No cerebral stroke was observed in the early postoperative period and no cerebrocardiovascular events were observed later. Conclusions:The short-term efficacy and safety of IAT in the treatment of RAO were satisfactory. The rescue time window might be prolonged.
6.High dose vitamin C inhibits proliferation of breast cancer cells through reducing glycolysis and protein synthesis.
Qingmei WANG ; Qianzi XU ; Anyi WEI ; Shishuo CHEN ; Chong ZHANG ; Linghui ZENG
Journal of Zhejiang University. Medical sciences 2019;48(3):296-302
OBJECTIVE:
To investigate the effects of high dose vitamin C (VC) on proliferation of breast cancer cells and to explore its mechanisms.
METHODS:
Human breast cancer cells Bcap37 and MDA-MB-453 were treated with VC at low dose (0.01 mmol/L), medium dose (0.10 mmol/L) and high dose (2.00 mmol/L). Cell proliferation was determined with CCK-8 assay, protein expression was evaluated by Western blot, and the secretion of lactic acid in tumor cells was detected by colorimetric method. Bcap37 cells were inoculated in nude mice, and tumor baring nude mice were intraperitoneally injected with high VC(4 g/kg, VC group, =5)or normal saline (control group, =5) for 24 d. Tumor weight and body weight were calculated.
RESULTS:
experiments demonstrated that high dose VC significantly inhibited cell proliferation in Bcap37 and MDA-MB-453 cells (all <0.01); the expressions of Glut1 and mTOR signaling pathway-related proteins were decreased (all <0.05); and the secretion of lactic acid was also markedly reduced (all <0.05). experiment showed that the tumor weight was decreased in mice treated with high-dose VC as compared with control group (<0.05), but no difference in body weights between two groups was observed.
CONCLUSIONS
High dose VC may inhibit proliferation of breast cancer cells both and through reducing glycolysis and protein synthesis.
Animals
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Ascorbic Acid
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pharmacology
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Breast Neoplasms
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drug therapy
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Cell Line, Tumor
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Cell Proliferation
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drug effects
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Glycolysis
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drug effects
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Humans
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Mice
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Mice, Nude
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Protein Biosynthesis
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drug effects
7.Investigation of Pretreatment Methods for Comprehensive Resource Utilization of Magnoliae Officinalis Cortex Dregs
Lukun XIAO ; Anyi ZHAO ; Jipeng DI ; An LIU ; Sha CHEN ; Jun ZHANG ; Aiping ZHANG ; Yan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(12):149-156
ObjectiveTo explore the pretreatment methods to promote the enzymatic digestion and extraction of active ingredients from Magnoliae Officinalis Cortex dregs(MOCD), and to provide a reference basis for the utilization of resource components in MOCD. MethodLiquid chromatography-mass spectrometry(LC-MS) was used for qualitative analysis of resource components in MOCD with an Agilent C18 reversed-phase column(3.0 mm×100 mm, 2.7 µm) at the flow rate of 0.4 mL·min-1, the mobile phase was water(A)-acetonitrile(B) for gradient elution(0-3 min, 25%-48%B; 3-6 min, 48%-59%B; 6-10 min, 59%-80%B; 10-20 min, 80%-90%B; 20-25 min, 90%B), electrospray ionization(ESI) was employed with negative ion mode scanning and scanning range of m/z 50-1 200. A high performance liquid chromatography(HPLC), which refered to the determination in the 2020 edition of Chinese Pharmacopoeia, was used for quantitative analysis of resource components in MOCD. Four kinds of pretreatment agents were used to separate the resource components from MOCD, and the mechanism of different pretreatment agents was investigated by field emission scanning electron microscopy(FESEM), X-ray powder diffraction(XRD) and Fourier transform infrared spectroscopy(FT-IR). ResultMagnolol, honokiol and lignocellulose were identified as the main resource components of MOCD by qualitative and quantitative analysis. Under the conditions of 1% NaOH, reaction temperature at 80 ℃ and reaction time of 60 min, the concentration of reducing sugar produced by the enzymatic hydrolysis was 32.18 g·L-1, which was 79.8% higher than that of the untreated MOCD. After adding tween-80, the enzymatic hydrolysis time was reduced to 1/3 of the original time, the concentration of reducing sugar was increased by 102.0%. And the total recovery of magnolol and honokiol in the pretreatment solution was 69.23%. ConclusionMagnolol, honokiol and lignocellulosic components in MOCD are valuable for development and utilization, the combination of alkaline pretreatment and tween-80 can realize the recovery and utilization of these three resource components, which can provide a new idea for comprehensive utilization of resource components in MOCD.
8.Application and Analysis of High-quality Evaluation for Jianwei Xiaoshi Tablets
Anyi ZHAO ; Lukun XIAO ; Sha CHEN ; Cong GUO ; Yuzhen LIU ; Zhiguang WANG ; Yan LIU ; An LIU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(21):138-146
Jianwei Xiaoshi tablets, as a common variety of Chinese patent medicine with "one product with many manufacturers", have many manufacturers and huge market sales. However, the phenomenon about uneven quality and discrepant price is prominent. Based on this, this study was carried out for the quality evaluation of Jianwei Xiaoshi tablets by applying the high-quality evaluation criteria with the quality as core for Chinese patent medicine, which was based on the full production cycle, from the multi-dimension including raw material selection, production process, quality control, post-marketing research and so on. The evaluation results showed that the quality evaluation scores of Jianwei Xiaoshi tablets from different manufacturers varied greatly (ranging from 35 to 66), indicating that the quality was significantly different. In the actual production, generally inadequate attention was paid to the quality of raw materials, and the quality of raw materials was insufficient with the score ratio of 43%, especially the poor consistency control of them. The role of good manufacturing practice was obvious, and the scores of production process were generally high with the average score ratio of 62%, and the maximum up to 80%. The technological advancement of the manufacturer was outstanding. The score ratio of quality control was only 31% that the internal quality standard of each manufacture almost stayed at the qualified line, which was equal to the national standard, and the consistency of products was insufficient. The post-marketing research was lacking with the score ratio of 37%. Manufacturers with high brand awareness and market share were upper scores, while the others lagged far behind. The results of this evaluation are in line with the overall prediction, which can provide a reference for the high-quality evaluation of Chinese patent medicine, and supply the scientific data for high-quality and high-price application.
9.Historical Evolution and Key Information Research on Pediatric Famous Classical Formula Yigongsan
Jiangmin SU ; Jun ZHANG ; Cong GUO ; Anyi ZHAO ; Liang JIANG ; Heng ZHANG ; Jipeng DI ; Sha CHEN ; Li LIU ; Yan LIU ; An LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):205-214
Yigongsan is derived from Xiaoer Yaozheng Zhijue written by QIAN Yi in the Northern Song dynasty, which is the No. 3 formula in the Catalogue of Ancient Famous Classical Formulas(The Second Batch of Pediatrics) released by the National Administration of Traditional Chinese Medicine(TCM) in September 2022, and it can be developed as a class 3.1 new TCM drug. By referring to ancient medical books and modern literature, this study conducted herbal textual research on Yigongsan from five aspects, including historical evolution, origin and processing, dosage conversion, usage and preparation methods, and functional application, then formed the key information table of this formula, in order to provide reference for the development of reference samples and preparations of Yigongsan. Based on the results of the study, it is recommended that Panax ginseng should be removed the basal part of stem(rhizoma), Poria cocos should be removed the peel, Citrus reticulata should be cut into shreds and Glycyrrhiza uralensis should be used. According to 4.13 g/Qian(钱), 1 g/slice for ginger, 3 g for each jujube and 300 mL/Zhan(盏), the doses of Ginseng Radix, Poria, Atractylodis Macrocephalae Rhizoma, Glycyrrhizae Radix et Rhizoma, Citri Reticulatae Pericarpium, Zingiberis Rhizoma Recens, Jujubae Fructus were 1.652, 1.652, 1.652, 1.652, 1.652, 5, 6 g, and the total amount was 19.26 g. The decocting method was to crush the medicinal materials into fine powder with 50-80 mesh, add 300 mL of water and decoct to 210 mL for each dose, then remove the dregs and take it warmly. This formula was recorded in ancient books as the main treatment for the cold-deficiency of spleen and stomach, and Qi stagnation in children with vomiting and diarrhea and lack of appetite. It has been flexibly applied by later generations of physicians, and is often used to treat anorexia, inflammation of the digestive tract, diarrhea and other diseases in children.
10.Quality Uniformity Analysis of 7 Commercially Available Yangyin Qingfei Preparations
Jiangmin SU ; Yongzhen LAO ; Sha CHEN ; Jun ZHANG ; Cong GUO ; Anyi ZHAO ; Jipeng DI ; An LIU ; Yan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):164-170
ObjectiveTo establish the fingerprint of seven commercially available Yangyin Qingfei preparations, to quantitatively analyze the index components, to evaluate their quality uniformity with multivariate statistical analysis, and to explore the quality differences among different dosage forms. MethodA total of 33 batches of commercially available 7 kinds of Yangyin Qingfei preparations were analyzed by high performance liquid chromatography(HPLC), the fingerprints were established and the common peaks were identified. Paeoniflorin, verbascoside, harpagoside, glycyrrhizic acid and paeonol were selected as the indicators of quality attributes to quantitatively analyze 33 batches of preparations. Based on the administration methods of Yangyin Qingfei preparations, the daily intake was calculated and the radar charts were poltted, and cluster analysis and principal component analysis were used to explore the quality differences of 7 kinds of Yangyin Qingfei preparations and the quality uniformity among different batches of the same dosage form. ResultThe similarity of fingerprints of 7 dosage forms was 0.248-0.956, suggesting that there were significant differences among different dosage forms of Yangyin Qingfei preparations, and a total of 15 common peaks were calibrated, of which peak 7, peak 8, peak 11, peak 13 and peak 15 were paeoniflorin, verbascoside, harpagoside, glycyrrhizic acid and paeonol, respectively. The radar plots showed that the average total daily intake of large honeyed pills and water honeyed pills was the highest, and the uniformity of pill components was better. The quality of 33 batches of samples was divided into poor quality and high quality by cluster analysis. Principal component analysis showed that the uniformity and dosage form of different dosage forms were significantly different, the oral liquid had the best quality homogeneity with the minimum dispersion. And the content of paeonol in different dosage forms was significantly different, which was the key component of quality control of Yangyin Qingfei preparations. ConclusionYangyin Qingfei large honeyed pills and water honeyed pills show high content and good uniformity, which are relatively preferred dosage forms. Different preparation processes have a great influence on the content of paeonol, and its quality control should be emphasized during production. This study provides a scientific method for the comparison of product quality of different dosage forms of traditional Chinese medicine prescriptions, which is helpful for the development of preferred dosage forms of different prescriptions, and provides a reference for efficient use of medication in the clinical practice.