2.Current situation and future development of SUMOylation in ophthalmology
Bo, LU ; Xin-Ling, WANG ; Qi-Chang, YAN
International Eye Science 2015;(8):1353-1357
SUMOylation is a post - translational modification consisting of covalent conjugation of ubiquitin - like proteins called small ubiquitin related modifier ( SUMO ) . SUMO modification has been shown to significantly alter protein activity, which can modulate protein stability, affect protein-protein interactions, and modify protein localization and trafficking. This process adds another layer of control in eukaryote gene expression, and it regulates both transcriptional activation and repression. This article reviews the current situation and future development of SUMOylation in ophthalmology.
3.Plasma ibuprofen enantiomers and their pharmacokinetics in Beagle dogs determined by HPLC.
Hongyan WANG ; Aiying KONG ; Bo YANG ; Liangping YAN ; Xin DI
Acta Pharmaceutica Sinica 2015;50(12):1607-12
A chiral high-performance liquid chromatography method was developed for the simultaneous determination of ibuprofen enantiomers in dog plasma. It was used to study the pharmacokinetics in the Beagle dog after intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen. Ketoprofen was chosen as the internal standard. After a simple precipitation using methanol as the precipitating solvent, both analytes and IS were separated on a Kromasil 100-5CHI-TBB chiral column (250 mm x4.6 mm, 5 μm) with isocratic elution using acetonitrile - 20 mmol x L(-1) phosphate buffer (pH 3.0, containing 5% methanol) (6 : 4) as the mobile phase. The detection wavelength was 220 nm. Liner calibration curves for both of the ibuprofen enantiomers were over the concentration range from 0.5 to 50 μg x mL(-1) with a lower limit of quantification of 0.5 μg x mL(-1), the accuracies were all in standard ranges. The intra- and inter- assay precisions were all below 7%. The recovery rate was 93.1% to 100.4%. The experiments proved that the method was simple, rapid and sensitive. It can be used in the quantitative determination of ibuprofen enantiomers in dog plasma. The method was used to determine the concentration of ibuprofen enantiomers in Beagle dog plasma after a single intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen (9 mg x kg(-1)) and the pharmacokinetics parameters were calculated based on the concentration-time curves. The C(max) of S-ibuprofen in Beagle dog plasma after a single intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen were 30.8 ± 4.7, 46.1 ± 5.9 and 20.0 ± 2.6 μg x mL(-1), respectively. In terms of the exposure of active ingredient, it revealed a significant difference between the administration of S-ibuprofen and the other two groups. The systematical R- to S- chiral inversion was discussed. Comparing the pharmacokinetic parameters at different doses, chiral inversion were 70.1% ± 36.6% and 76.4% ± 36.2%, respectively, after intravenous administration of racemic- and R-ibuprofen. This study provides a theoretical basis for the safety of ibuprofen formula of injection drug.
4.Plasma ibuprofen enantiomers and their pharmacokinetics in Beagle dogs determined by HPLC.
Hong-yan WANG ; Ai-ying KONG ; Bo YANG ; Liang-ping YAN ; Xin DI
Acta Pharmaceutica Sinica 2015;50(12):1607-1612
A chiral high-performance liquid chromatography method was developed for the simultaneous determination of ibuprofen enantiomers in dog plasma. It was used to study the pharmacokinetics in the Beagle dog after intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen. Ketoprofen was chosen as the internal standard. After a simple precipitation using methanol as the precipitating solvent, both analytes and IS were separated on a Kromasil 100-5CHI-TBB chiral column (250 mm x4.6 mm, 5 μm) with isocratic elution using acetonitrile - 20 mmol x L(-1) phosphate buffer (pH 3.0, containing 5% methanol) (6 : 4) as the mobile phase. The detection wavelength was 220 nm. Liner calibration curves for both of the ibuprofen enantiomers were over the concentration range from 0.5 to 50 μg x mL(-1) with a lower limit of quantification of 0.5 μg x mL(-1), the accuracies were all in standard ranges. The intra- and inter- assay precisions were all below 7%. The recovery rate was 93.1% to 100.4%. The experiments proved that the method was simple, rapid and sensitive. It can be used in the quantitative determination of ibuprofen enantiomers in dog plasma. The method was used to determine the concentration of ibuprofen enantiomers in Beagle dog plasma after a single intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen (9 mg x kg(-1)) and the pharmacokinetics parameters were calculated based on the concentration-time curves. The C(max) of S-ibuprofen in Beagle dog plasma after a single intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen were 30.8 ± 4.7, 46.1 ± 5.9 and 20.0 ± 2.6 μg x mL(-1), respectively. In terms of the exposure of active ingredient, it revealed a significant difference between the administration of S-ibuprofen and the other two groups. The systematical R- to S- chiral inversion was discussed. Comparing the pharmacokinetic parameters at different doses, chiral inversion were 70.1% ± 36.6% and 76.4% ± 36.2%, respectively, after intravenous administration of racemic- and R-ibuprofen. This study provides a theoretical basis for the safety of ibuprofen formula of injection drug.
Animals
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Chromatography, High Pressure Liquid
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Dogs
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Ibuprofen
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blood
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pharmacokinetics
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Stereoisomerism
5.Investigation and molecular identification of Anisakis infection in marine fish sold in Fuxin, Liaoning Province
REN Yan-yan ; DU Bo ; GAI Nan-nan ; XIU Min ; LIU Wen-xin
China Tropical Medicine 2023;23(5):489-
Abstract: Objective To investigate the infection of Anisakis in marine fish sold in Fuxin, and conduct molecular identification and evolutionary tracing of third-stage larvae to determine Anisakis species. Methods From 2018 to 2021, marine fish sold in the market were collected randomly, and the third stage larvae of Anisakis were detected in marine fish sold in the market by direct dissection, and the morphological characteristics were used to preliminarily identify species by microscopy; the total DNA was extracted, the internal transcribed spacer sequence of the ribosomal DNA of Anisakis was amplified, and the sequence alignment and evolution analysis were carried out. Results A total of 289 market-sold sea fish samples of marine fish sold in the market were dissected and 84 samples of Anisakis were detected with a detection rate of 29.1%, of which the infection rates of hairtail and small yellow croaker were higher, at 41.4% and 41.2%, respectively. BLAST comparison of 28 sequences revealed eight species of anisakids, including Anisakis pegreffii, Anisakis simplex, Anisakis typical, Raphidascaris trichiurid, Contracaecum muraenesoxi, Hysterothylcaium zhoushanensis, Hysterothylacium amoyense and Hysterothylcaium fabri,belonging to the genera Anisakis and Hysterothylacium. The phylogenetic tree constructed from 28 sequences generally formed two topological branches, with Anisakis pegreffii, Anisakis simplex, and Anisakis typical forming three separate clusters as the topology branch of Anisakis genus. However, meanwhile, Hysterothylacium, Contracaecum, and Raphidascaris formed a separate topological branch. Conclusions The marine fish sold in Fuxin City have severe anisakid infection, with a wide variety of anisakid species, the dominant species being Anisakis pegreffii.
7.Evaluation indexes for the viability of umbilical cord-derived mesenchymal stem cells before transplantation
Xin LEI ; Yan CHEN ; Jianlin ZHANG ; Lei CUI ; Yuhu NIU ; Bo NIU
Chinese Journal of Tissue Engineering Research 2013;(32):5847-5854
BACKGROUND:Umbilical cord-derived mesenchymal stem cel s are gaining more attention in clinical treatments. Cel viability prior to transplantation has a direct impact on clinical prognosis. Despite trypan blue staining is a widely performed procedure to assess the viability of umbilical cord-derived mesenchymal stem cel s, it cannot reflect the functional capacity of those cel s accurately because of some subjective factors. OBJECTIVE:To explore sensitive and accurate assay for the functions of umbilical cord-derived mesenchymal stem cel s. METHODS:Human umbilical cord-derived mesenchymal stem cel s were isolated and cultured in vitro. Cultured umbilical cord-derived mesenchymal stem cel s were preserved in 0.9%saline for 0, 2, 4 and 6 hours at 4 ℃. Various methods (trypan blue staining, AnnexinV-PI, terminal deoxynucleotidyl transferase dutp nick end labeling, cel counting kit-8, live-dead assay, cel adherent assay) were used to determine the viability of post-storage umbilical cord-derived mesenchymal stem cel s, and the results were compared with colony-forming efficiency, a measure of cel function. RESULTS AND CONCLUSION:Human umbilical cord-derived mesenchymal stem cel s cultured in vitro showed a spindle shape and attached growth, the third-generation umbilical cord-derived mesenchymal stem cel s were positive for CD29, CD44, CD105, and negative for CD 34 and CD 45. Umbilical cord-derived mesenchymal stem cel s incubated in the adipogenic and osteogenic medium were both positive. Cel viability measured with trypan blue correlated moderately with colony-forming efficiency, while the percentage of viable cel s measured with other methods correlated better with colony-forming efficiency, among which adherent assay was the most obvious. It is proved that cel adherent assay-measured viability is the most accurate indicator.
8.Oxidative stress effect on viability of umbilical cord-derived mesenchymal stem cells in storage solution of transplantation
Yuhu NIU ; Yan CHEN ; Jianlin ZHANG ; Xin LEI ; Yanting DONG ; Lei CUI ; Bo NIU
Chinese Journal of Tissue Engineering Research 2013;(32):5785-5792
BACKGROUND:The viability of human umbilical cord-derived mesenchymal stem cel s is often declined with the commonly used transplantation storage solution in clinics, which may influence the therapeutic effects of cel ular transplantation. However, reasons for this are stil unknown. OBJECTIVE:To investigate the role of oxidative stress in the reduction of human umbilical cord-derived mesenchymal stem cel s viability in the storage process during clinical transplantation and to observe the effects of radical scavenger on the results. METHODS:Human umbilical cord-derived mesenchymal stem cel s were harvested and cultured in normal saline for 0, 2, 4 and 6 hours at room temperature. Intracel ular reactive oxygen levels were detected at those time points. Antioxidant enzyme activities and levels of malondialdehyde were measured to determine the intracel ular oxidative stress levels after storage. Cel adhesion rate changes were retested after adding N-acetyl cysteine to the storage solution. RESULTS AND CONCLUSION:The reactive oxygen levels in human umbilical cord-derived mesenchymal stem cel s were increased significantly after normal saline storage and levels of malondialdehyde were increased in a time-dependent manner. Activities of superoxide dismutase, catalase and glutathione peroxidase were al reduced. Addition of N-acetyl cysteine into the storage medium decreased the reactive oxygen levels and improved the human umbilical cord-derived mesenchymal stem cel s viabilities. Experimental findings indicate that, increased reactive oxygen species in human umbilical cord-derived mesenchymal stem cel s is one of the reasons for reduced cel viability. Adding the radical scavenger N-acetyl cysteine can improve the storage effects of human umbilical cord-derived mesenchymal stem cel s.
9.Curative effects of excimer laser corneal refractive surgery for hyperopic anisometropic amblyopia
Xin-Gang, NIE ; Yan-Xia, HUANG ; Ying-Ying, HE ; Bo-Li, DONG
International Eye Science 2017;17(7):1381-1383
AIM: To investigate curative effects of excimer laser corneal refractive surgery for adults or older adolescent with hyperopic anisometropic amblyopia.METHODS: From March 2014 to March 2016, we selected 26 cases 26 eyes of adults or older adolescent with hyperopic anisometropic amblyopia in our hospital.All eyes underwent laser in situ keratomileusis, observed for the uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), diopter and stereopsis.RESULTS: At the end of the follow-up, the patient`s spherical equivalent and anisometropia were 1.47±0.51D and 1.15±0.22D, were significantly lower than that before operation (P<0.05).At the end of the follow-up, the distance and near UCVA and BCVA were 0.26±0.13 and 0.23±0.09, 0.42±0.09 and 0.31±0.16, which were significantly higher than those before operation (P<0.05).At the end of follow-up, the visual function of the patients was significantly improved (P<0.05), the rate of postoperative visual function < 100 eyes was 23%.CONCLUSION: In adult or older adolescent with hyperopic anisometropic amblyopia, excimer laser corneal refractive surgery has a certain effect.
10.lnhibition effect of AG490 on high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells
Li, FENG ; Liang, ZHANG ; Xiao, HAN ; Bo, LU ; Xin-Ling, WANG ; Qi-Chang, YAN
International Eye Science 2014;(12):2122-2126
AlM:To study the relationship between JAK-STAT pathway and epithelial - mesenchymal transition in human lens epithelial cells. Meanwhile, the function of AG490 as a JAK inhibitor was also demonstrated in this article.
METHODS:Human lens epithelial cells SRA01/04 ( LECs ) were treated by low concentration of glucose (5. 5mmol/L). High concentration of glucose (30. 5mmol/L) was used to treat the cells in order to form the high glucose model. According to adding AG490 or not, cells were divided into the control group and the experimental group, appropriate concentration 10ü mol/L and 50ü mol/L of AG490 were chosen and acting time of 6, 12, 24, 48h were selected. Effect of AG490 on cell migration was measured by wound healing test. The expression of TGF-β1 , FN,α-SMA mRNA were examined by RT-PCR.
RESULTS:With the prolonged acting time ( 6, 12, 24 and 48h), cell activity increased in the HG group, as well as more expression of TGF-β1 , FN,α-SMA mRNA were detected compared to the LG group (P<0. 05). ln AG490 group, the cell migration activity and expression of TGF-β1 , FN,α-SMA mRNA decreased compared to the HG group (P<0. 05).
CONCLUSlON:JAK-STAT pathway takes part in high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells. The mechanism is that it impacts the transcriptional expression of TGF- β1 and extracellular matrix. AG490, a JAK inhibitor, inhibits high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells, And the inhibition enhances with the increasing concentration of AG490.