1.In Vitro and in Vivo Evaluation of Scutellarin-phospholipid Complex Nanoemulsion and Analysis of Its Activity in Ameliorating LPS-induced Vascular Endothelial Injury
Tian LUO ; Zhiyong HE ; Xiangjun MAO ; Xue LIU ; Jinggang HE ; Yuan ZHI ; Xiangchun SHEN ; Qianli XU ; Ling TAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):159-168
ObjectiveTo evaluate some properties of scutellarin-phospholipid complex nanoemulsion(SCU-PC-NE), such as release, cell uptake and tissue distribution, and to investigate its effect on ameliorating lipopolysaccharide(LPS)-induced vascular endothelial injury. MethodSCU-PC-NE was prepared by weighting SCU-PC, ethyl oleate, Kolliphor HS15, 1,2-propylene glycol(50, 400, 514.3, 85.7 mg), respectively. And the appearance of SCU-PC-NE was observed by transmission electron microscope, the average paticle size and Zeta potential were measured by nanopotential particle size analyzer. The cumulative release of SCU-PC-NE in vitro was measured by dynamic dialysis, thiazolyl blue(MTT) colorimetric assay was used to investigate the effect of SCU-PC-NE on the viability of human umbilical vein endothelial cells(HUVECs), the inverted fluorescence microscope and flow cytometry were used to investigate cell uptake of HUVECs by SCU-PC-NE in vitro using coumarin 6 as a fluorescent probe, the tissue distribution of DiR/SCU-PC-NE labeled by near infrared fluorescent dyes was obeserved by small animal in vivo imaging system. The inflammation injury model was established by co-incubation with LPS(1 mg·L-1) and HUVECs, the effect of SCU-PC-NE on the levels of interleukin(IL)-1β and IL-6 were determined by enzyme-linked immunosorbent assay(ELISA), 18 Kunming male mice were randomly divided into blank group, model group, blank preparation group(equivalent to high dose group), SCU group and SCU-PC-NE low and high dose groups(5, 10 mg·kg-1), 3 mice in each group, and the drug administration groups were administered once in the tail vein at the corresponding dose every 48 h, equal volume of normal saline was given to the blank group and the model group, and the drug was administered for 4 consecutive times. Except for the blank group, the endothelial inflammatory injury was induced by intraperitoneal injection of LPS(10 mg·kg-1) at 12 h before the last administration in each group. Hematoxylin-eosin(HE) staining was used to investigate the effect of SCU-PC-NE on the histopathological changes in the thoracic aorta of mice. ResultThe appearance of SCU-PC-NE displayed pale yellow milky light, mostly spherical with rounded appearance and relatively uniform particle size distribution, with the average particle size of 35.31 nm, Zeta potential of 7.23 mV, and the encapsulation efficiency of 75.24%. The cumulative release in vitro showed that SCU-PC-NE exhibited sustained release properties compared with SCU. The cell viability of SCU-PC-NE was >90% at a concentration range of 1.05-8.4 mg·L-1. The results of cellular uptake experiments showed that the cellular uptake ability of SCU-PC-NE was significantly enhanced when compared with the SCU group(P<0.01). Compared with normal mice, the results of tissue distribution showed that the fluorescence intensity of DiR/SCU-PC-NE was significantly enhanced in the spleen, kidney, brain and thoracic aorta of mice at different time points after intraperitoneal injection of LPS(P<0.05, P<0.01), especially in thoracic aorta. ELISA results showed that the levels of IL-1β and IL-6 in the model group were significantly increased when compared with the blank group(P<0.05, P<0.01), and compare with the model group, all administration groups significantly down-regulated IL-1β level, with the strongest effect in the SCU-PC-NE high-dose group(P<0.01), and all administration groups significantly down-regulated IL-6 level, with the strongest effect in the SCU-PC-NE low-dose group(P<0.05). Compare with the blank group, the results of HE staining showed that the endothelial cells were damaged, the elastic fibers were broken and arranged loosely in the model group, although similar vascular injury could be observed in the blank preparation group, SCU group and SCU-PC-NE low-dose group, the vascular endothelial damage was significantly reduced in the high-dose group of SCU-PC-NE, which had a better effect than that in the SCU group. ConclusionSCU-PC-NE can promote the uptake of drugs by endothelial cells and effectively enriched in the site of vascular endothelial injury caused by LPS, suggesting that it has a protective effect on vascular endothelial injury and is a good carrier of SCU.
2.Wireless closed-loop deep brain stimulation using microelectrode array probes
JIA QIANLI ; LIU YAOYAO ; LV SHIYA ; WANG YIDING ; JIAO PEIYAO ; XU WEI ; XU ZHAOJIE ; WANG MIXIA ; CAI XINXIA
Journal of Zhejiang University. Science. B 2024;25(10):803-823
Deep brain stimulation(DBS),including optical stimulation and electrical stimulation,has been demonstrated considerable value in exploring pathological brain activity and developing treatments for neural disorders.Advances in DBS microsystems based on implantable microelectrode array(MEA)probes have opened up new opportunities for closed-loop DBS(CL-DBS)in situ.This technology can be used to detect damaged brain circuits and test the therapeutic potential for modulating the output of these circuits in a variety of diseases simultaneously.Despite the success and rapid utilization of MEA probe-based CL-DBS microsystems,key challenges,including excessive wired communication,need to be urgently resolved.In this review,we considered recent advances in MEA probe-based wireless CL-DBS microsystems and outlined the major issues and promising prospects in this field.This technology has the potential to offer novel therapeutic options for psychiatric disorders in the future.
3.Novel mutation c.1210-3C > G in cis with a poly-T tract of 5T affects CFTR mRNA splicing in a Chinese patient with cystic fibrosis.
Xinyue ZHAO ; Keqiang LIU ; Wenshuai XU ; Meng XIAO ; Qianli ZHANG ; Jiaxing SONG ; Keqi CHEN ; Yaping LIU ; Xinlun TIAN ; Kai-Feng XU ; Xue ZHANG
Frontiers of Medicine 2022;16(1):150-155
Cystic fibrosis (CF) is a rare autosomal recessive disease with only one pathogenic gene cystic fibrosis transmembrane conductance regulator (CFTR). To identify the potential pathogenic mutations in a Chinese patient with CF, we conducted Sanger sequencing on the genomic DNA of the patient and his parents and detected all 27 coding exons of CFTR and their flanking intronic regions. The patient is a compound heterozygote of c.2909G > A, p.Gly970Asp in exon 18 and c.1210-3C > G in cis with a poly-T of 5T (T5) sequence, 3 bp upstream in intron 9. The splicing effect of c.1210-3C > G was verified via minigene assay in vitro, indicating that wild-type plasmid containing c.1210-3C together with T7 sequence produced a normal transcript and partial exon 10-skipping-transcript, whereas mutant plasmid containing c.1210-3G in cis with T5 sequence caused almost all mRNA to skip exon 10. Overall, c.1210-3C > G, the newly identified pathogenic mutation in our patient, in combination with T5 sequence in cis, affects the CFTR gene splicing and produces nearly no normal transcript in vitro. Moreover, this patient carries a p.Gly970Asp mutation, thus confirming the high-frequency of this mutation in Chinese patients with CF.
China
;
Cystic Fibrosis/genetics*
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Cystic Fibrosis Transmembrane Conductance Regulator/genetics*
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Humans
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Mutation
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Poly T
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RNA, Messenger/genetics*
4.The role of three-dimensional speckle tracking imaging in the diagnosis of immunoglobulin light-chain cardiac amyloidosis with normal left ventricular ejection fraction
Changhui LEI ; Lei ZUO ; Yan WANG ; Xiaoli ZHU ; Mengyao ZHOU ; Qianli YANG ; Hanxi XU ; Liwen LIU
Chinese Journal of Ultrasonography 2020;29(3):213-218
Objective:To explore the value of three-dimensional speckle tracking imaging (3D-STI) in the diagnosis of immunoglobulin light-chain cardiac amyloidosis(AL-CA) patients with normal left ventricular ejection fraction (LVEF).Methods:A total of 92 consecutive patients diagnosed with systemic immunoglobulin light chain amyloidosis(sAL) and with normal LVEF from October 2014 to January 2018 in Xijing Hospital were enrolled.Based on the diagnostic criteria of cardiac involvement, the patients were divided into AL-CA group (52 cases) and immunoglobulin light chain amyloidosis (AL) group (40 cases). The clinical data and serological markers of the patients were collected, the conventional echocardiography and full-volume three dimensional dynamic images were acquired, left ventricular global longitudinal strain (GLS), global radial strain (GRS), global circumferential strain (GCS), and global area strain (GAS) were analyzed using off-line TomTec software. The differences between the two groups were compared.Results:Compared with the AL group, the NT-proBNP of AL-CA group was significantly higher ( P<0.05) and there were no significant differences of the other serological indexes between the two groups(all P>0.05). Compared with the AL group, the maximal left ventricular wall thickness, left ventricular mass index, left atrial volume index, and E/e′ in the AL-CA group were significantly increased (all P<0.05). There were no significant differences of other conventional echocardiographic measurements between the two groups(all P>0.05). Compared with the AL group, GLS, GAS, and GRS were significantly lower in AL-CA group (all P<0.05); but there was no significant difference of GCS between the two groups( P>0.05). The ROC curve analysis showed that the cut-off values discriminating cardiac involvement were 16.09% for GLS, 36.54% for GAS and 31.90% for GRS. Conclusions:3D-STI measurements of left ventricular myocardial mechanics could detect cardiac involvement in patients with sAL amyloidosis, and provides a new method for diagnosis of AL-CA.
5.Study on the Improvement of Quality Standard for Keqing Capsules
Xue LIU ; Hongyan QIU ; Jing PENG ; Chunyan LIU ; Shuyue QU ; Ying MA ; Qianli XU ; Xiangchun SHEN ; Ling TAO ; Xiangjun MAO
China Pharmacy 2020;31(5):595-600
OBJECTIVE:To optimi ze and improve the quality standard for Keqing capsules. METHODS :According to general rule 0502 method stated in 2015 edition of Chinese Pharmacopeia (part Ⅳ),TLC method was used to identify Reineckia carnea and Morus alba in Keqing capsules [the developing solvents were dichloromethane-ethyl acetate-formic acid (10 ∶ 4 ∶ 0.2,V/V/V) and ethyl acetate-carbinol-ammonia (12 ∶ 2 ∶ 1,V/V/V),respectively]. The contents of morphine and codeine phosphate in Keqing capsules were determined by HPLC. The determination was performed on XBridge C 18 column with mobile phase consisted of acetonitrile-0.01 mol/L potassium dihydrogen phosphate aqueous solution (pH value adjusted to 2.7 with 5% phosphoric acid solution)(5 ∶ 95,V/V)at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm,and the column temperature was 35 ℃. The sample size was 10 µL. RESULTS :In TLC of R. carnea and M. alba in samples ,same color spots were shown in the correspon ding positions of reference substance chromatogram without interference from negative control. The linear range of morphine and codeine phosphate were batches of Keqing capsules were 0.97-1.37,0.16-0.37 mg/g,respectively. CONCLUSIONS :TLC identification method for R. carnea and M. alba ,as well as HPLC content determination method for morphine and codeine phosphate in Keqing capsules are established;the method is simple ,accurate and reliable with strong specificity ,which improves the quality standard of Keqing capsules.
6.Developmentand application of maker education in orthodontics course for undergraduate
Yifan HE ; Qianli LI ; Xiaolei LI ; Jingyi XUE ; Cheng ZHANG ; Yue XU
Chinese Journal of Medical Education Research 2019;18(4):385-389
In order to stimulate students' interest in orthodontics and develop their competency in self-directed learning,innovation and practice,we introduced "maker education" into the orthodontics course for undergraduate.In the early stage,the teaching materials were organized based on fragmentation of learning,and the "maker group" is the basic unit.In class,the topics were discussed in the form of flipped classroom.Finally,a series of orthodontic experiments were carried out.This article can provide new ways to improve the quality of orthodontics education.
7.An echocardiographic study of left ventricular torsion in patients with latent obstructive hypertrophic cardiomyopathy
Lei ZUO ; Jing WANG ; Xin MENG ; Changhui LEI ; Qianli YANG ; Nan KANG ; Fan YANG ; Lei XU ; Liwen LIU
Chinese Journal of Ultrasonography 2019;28(4):277-282
Objective To explore the changes of left ventricular torsion function in patients with latent obstructive hypertrophic cardiomyopathy ( HCM ) ,and provide quantitative informations for clinical evaluation of cardiac function . Methods A total of 49 consecutive patients with HCM without left ventricular outflow tract obstruction at rest were enrolled . All subjects underwent exercise stress echocardiography . After exercise left ventricular outflow tract pressure gradient ( LVO T‐PG ) ≥30 mm Hg was positive for exercise stress test ( latent obstruction) ,w hile LVO T‐PG< 30 mm Hg was negative for exercise stress test ( non‐obstruction) . An ultrasound system obtained two‐dimensional ultrasound images of resting and moving peaks . The global longitudinal strain ( GLS ) ,global circumferential strain ( GCS ) , global radial strain ( GRS) of the left ventricle 16 segments and left ventricular rotation ,twist were analysis using off‐line EchoPAC software . T he differences of the above parameters were compared between the two groups . Results T here were no significant differences in GLS ,GRS ,GCS and Rotation‐B between the two groups in resting and peak period of exercise ( all P > 0 .05 ) ,GRS in both groups were significantly increased compared with that before exercise ( all P < 0 .05 ) . Compared with the negative exercise stress group ,the left ventricular twist and Rotation‐A were significantly increased in resting and peak period of exercise in the positive exercise stress test group( all P <0 .05) . Compared with before exercise ,Rotation‐A and left ventricular twist were significantly decreased in the positive exercise stress test group ( all P <0 .05) ,while no significantly difference was found in the negative exercise stress group ( all P > 0 .05 ) . Conclusions Left ventricular torsion function is significantly changed in rest and after exercise in latent obstructive HCM patients ,providing valuable quantitative information for clinical comprehensive evaluation of cardiac function .
8.Effect of the chemoprotectant tempol on anti-tumor activity of cisplatin.
Shuangyan YE ; Sisi ZENG ; Mengqiu HUANG ; Jianping CHEN ; Xi CHEN ; Pengfei XU ; Qianli WANG ; Wenwen GAO ; Bingsheng YANG ; Bingtao HAO ; Wenhuan HUANG ; Qiuzhen LIU
Journal of Southern Medical University 2019;39(8):883-890
OBJECTIVE:
To investigate the effect of the chemoprotectant tempol on the anti-tumor activity of cisplatin (DDP).
METHODS:
The cellular toxicity of tempol in human colon cancer SW480 cells and mouse colon cancer CT26 cells were evaluated using MTT and cell counting kit-8 assays. CalcuSyn software analysis was used to determine the interaction between tempol and DDP in inhibition of the cell viability. A subcutaneous homograft mouse model of colon cancer was established. The mice were randomly divided into control group, tempol group, cisplatin group and tempol + DDP treatment group with intraperitoneal injections of the indicated agents. The tumor size, body weight and lifespan of the mice were measured, and HE staining was used to analyze the cytotoxic effect of the agents on the kidney and liver. Immunohistochemistry and Western blotting were performed to detect the expression of Bax and Bcl2 in the tumor tissue, and TUNEL staining was used to analyze the tumor cell apoptosis. The level of reactive oxygen species (ROS) in the tumor tissue was determined using flow cytometry.
RESULTS:
Tempol showed inhibitory effects on the viability of SW480 and CT26 cells. CalcuSyn software analysis showed that tempol had a synergistic anti-tumor effect with DDP (CI < 1). In the homograft mouse model, tempol treatment alone did not produce obvious anti-tumor effect. HE staining showed that the combined use of tempol and DDP alleviated DDP-induced fibrogenesis in the kidneys, but tempol also reduced the anti-tumor activity of DDP. Compared with the mice treated with DDP alone, the mice treated with both tempol and DDP had a significantly larger tumor size ( < 0.01) and a shorter lifespan ( < 0.05). Tempol significantly reversed DDP-induced expression of Bax and Bcl2 in the tumor tissue and tumor cell apoptosis ( < 0.001), and obviously reduced the elevation of ROS level in the tumor tissue induced by DDP treatment ( < 0.05).
CONCLUSIONS
Tempol can attenuate the anti-tumor effect of DDP while reducing the side effects of DDP. Caution must be taken and the risks and benefits should be carefully weighed when considering the use of tempol as an anti-oxidant to reduce the toxicities of DDP.
Animals
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Antineoplastic Agents
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Antioxidants
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Apoptosis
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Cell Line, Tumor
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Cell Proliferation
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Cisplatin
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Cyclic N-Oxides
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pharmacology
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Drug Resistance, Neoplasm
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Humans
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Mice
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Spin Labels
9.Quality standard for Cyclocarya paliurus
Linlin WU ; Fang WANG ; Xiangjun MAO ; Qianli XU
Chinese Traditional Patent Medicine 2017;39(4):745-750
AIM To establish the quality standard for Cyclocarya paliurus (Batal.) ljinsk..METHODS The contents of water,ash and extract in twelve batches of samples were determined.TLC and HPLC were adopted in the qualitative identification and quantitative determination of quercetin and kaempferol,respectively,and phenol-sulfuric acid method was used for the polysaccharide content assessment.RESULTS The average contents of water,total ash,acid-insoluble ash,water-soluble extract and alcohol-soluble extract were 11.05%,5.81%,1.70%,11.25% and 10.16%,respectively.The clear TLC spots demonstrated their strong specificity.Quercetin and kaempferol showed good linear relationships within the ranges of 0.004 953-0.022 29 mg/mL and 0.005 748-0.028 74 mg/mL (r =0.999 9),whose average recoveries were 97.1% (RSD =2.59%) and 97.9% (RSD =2.86%),respectively.Polysaccharide showed a good linear relationship within the range of 0.021 22-0.095 58 mg/mL,whose average recovery was 97.2% (RSD =2.42%).The contents of three constituents in various batches of samples showed obvious differences.CONCLUSION In C.paliurus,the contents of water,total ash,acid-insoluble ash,water-soluble extract and ethanol-soluble extract should not be more than 13.0%,7.0%,2.0%,13.5% and 12.0%,while those of quercetin,kaempferol and polysaccharide (calculated by dry product) should not be less than 0.040%,0.070% and 0.60%,respectively.
10.Clinical value of noninvasive cardiac index test in the evaluation of neonatal congenital heart disease complicated with heart failure
Yonghua YUAN ; Aimin ZHANG ; Xuehua HE ; Jun XU ; Furong HUANG ; Liping LIU ; Zhenyu LIU ; Xiaohui XIA ; Mei LV ; Aitong QIANLI ; LI ZHU
Journal of Clinical Pediatrics 2017;35(10):747-750
Objective To explore the clinical value of the monitoring of electronic cardiac index (CI) in the evaluation of neonatal congenital heart disease complicated with heart failure. Methods Sixty neonates with congenital heart disease treated in neonatal department from March 1, 2016 to December 30, 2016 were selected, and divided into severe group (n=11), moderate group (n=15), mild group (n=34), and no heart failure group (n=10) according to the modified Ross heart failure score. CI was measured by electronic force measurement. Left ventricular ejection fraction (LVEF) and pulmonary arterial pressure (PAP) were measured by echocardiography. Venous blood sampling was collected to detect the N-terminal type B brain natriuretic peptide (NT-proBNP). Results The neonates in the severe group were mainly under 2-week-old, while those in the mild group and the moderate group were more than 2-week-old. The differences of CI, LVEF, NT-proBNP, and PAP among the groups were statistically different. The CI and LVEF values were lowest in the severe group, followed by moderate group and mild group, and the highest in no heart failure group. The NT-proBNP and PAP values were the highest in the severe group, followed by moderate group and mild group, and the lowest in no heart failure group. Correlation analysis showed that CI was positively correlated with LVEF (r=0.845, P<0.001), and negatively correlated with NT-proBNP (r=-0.886, P<0.001); CI and PAP were weakly negatively correlated (r=-0.595, P<0.001). Conclusions CI reflects the degree of heart failure to some extent and has some clinical value.

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