1.Changes of MMP-3 and MMP-9 expressions on human trabecular cells following the exposure of laser associated with selective laser trabeculoplasty
Hongyang, ZHANG ; Yiming, YE ; Yangfan, YANG ; Minbin, YU
Chinese Journal of Experimental Ophthalmology 2015;33(3):227-231
Background Selective laser trabeculoplasty (SLT) can increase the outflow of aqueous humor and reduce the intraocular pressure of patients with open angle glaucoma,but its mechanism is unknown.To investigate the mechanism of SLT would improve the therapeutic effect of SLT.The aqueous outflow resistance in trabecular meshwork was affected by the extracellular matrix (ECM).Matrix metalloproteinase-3 (MMP-3) and MMP-9 were closely related to ECM degradation in trabecular meshwork.Objective This study was to observe the effects of SLT on the expressions MMP-3 and-9 in human trabecular ceils in vitro.Methods Immortallized human trabecular cells were cultured with pigment particles mixed suspension for 16 hours and incubated overnight.Then the cells were irradiated with Q switch frequency doubling 532 nm Nd:YAG laser to establish SLT-effective cells with the energy of 0.2 mJ,spot diameter of 400 μm and pulse duration of 3 ns.The expressions of MMP-3 mRNA and MMP-9 mRNA in the cells were detected by fluorescence quantitative real time PCR before and 1 hour and 4,8,12,28 hours after exposure of laser.The concentrations of MMP-3 and MMP-9 in the medium were assayed using ELISA 1 hour and 4,8,12,28 hours after exposure of laser and compared between the non-irradiation group and the irradiation group.Results The relative expressing levels of MMP-3 mRNA were 1.00,1.32±0.12,2.08±0.05,2.34±0.04,2.77± 0.05 and 2.49±0.27 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of laser SLT,showing a significant difference among the groups (F =15.966,P=0.007),and relative expressing levels of MMP-3 mRNA were significantly higher in various time points after laser irradiation than those of the non-irradiation group (all at P<0.05).The relative expressing levels of MMP-9 mRNA were 1.00,0.91 ±0.10,1.27 ± 0.07,1.46±0.07,1.69±0.09 and 0.87±0.09 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of SLT,which was considerably different among the groups (F =30.526,P =0.005),and significant increased values were seen in the 4,8 and 12 hours after irradiation compared with the non-irradiation group (all at P<0.05),with highest expression in the irradiation for 12-hour group.The concentrations of MMP-3 and MMP-9 proteins in medium were significantly increased in various time points after laser exposure in comparison with the control group (all at P<0.05).Conclusions The expressions of MMP-3 and MMP-9 in human trabercuolar cells upregulate and the secretion ability of human trabecular cells to MMP-3 and MMP-9 proteins improves in early stage of SLT in vitro.However,these procedures gradually diminish with the lapse of time.
2.Establishment of selective laser trabeculoplasty effect model in human trabecular cell in vitro
Hongyang, ZHANG ; Yiming, YE ; Yangfan, YANG ; Minbin, YU
Chinese Journal of Experimental Ophthalmology 2014;32(2):107-113
Background Selective laser trabeculoplasty (SLT) plays lowing-intraocular pressure (IOP)effect by irradiating pigmented trabecular cells selectively using 532 nm Nd:YAG laser.However,its affect pattern is not fully known.Objective This study was to establish a human trabecular cells (HTCs) model of SLT effect,and to observe the morphological changes of HTCs after they were exposed to different energies of SLT.Methods Immortalized HTCs were incubated in DMEM/F12 with pigmental granules for 16 hours (overnight) to prepare pigmented trabecular cells.The cells were irradiated with modified laser lens with the spot of 400 μm and emitting duration of 3 ns.The energy scale of 0.2 mJ was set for standardized energy and 0.1 mJ was used as the low energy.The morphology and ultrastructure of the cells were examined under the light microscope and transmission electron microscope 1 hour,4,8,12,24 hours after irradiation.Trypan blue staining and TUNEL fluorescence staining were used to evaluate the death and apoptosis of the cells after laser irradiation.Results The brown pigment particles were seen in cytoplasm around nuclei of trabecular cells after cultured for 16 hours.After laser irradiation,there was no obvious change in the shape of nonpigmented trabcular cells,but a 400 μm spot was found in the pigmented trabecular cells under the light microscope,and the pigmented particles released as the cell disruption.After 0.2 mJ laser irradiation,cell loss zone was seen at the center of the laser spot,and the positive cells for trypan blue and TUNEL staining were found;while the abnormal manifestations were slight after the 0.1 mJ irradiation.With the lapse of the time,the number of positive staining cells was gradually decreased.Twenty-four hours after laser irradiation,proliferative trabecular cells emerged and migrated to the center area of laser spot.Statistical analysis showed that the numbers of the positive cells for trypan blue and TUNEL staining were significantly reduced in the lower energy group in comparison with the standardized energy group at various time points (all at P<0.001),and those of both energy groups were gradually reduced with lapse of time with the significant differences between any two time points (all at P<0.01).Conclusions An in vitro laser effect model of HTCs can be established by exposing pigmented HTCs to SLT laser.After exposed to SLT,the gasification,necrosis-like death,apoptosis-like change appear at the laser center.The laser destroyed cells can be replaced by proliferative cells with the lapse of time.Low-energy laser irradiation cause a similar morphological change of the cells to high-energy irradiation,but the number of abnormal cells is much less.
3.Effect of Chaihu Shugan Powder on the content of 5-hydroxytryptamine and the expression of tryptophan hydroxylase 2 in depression rats
Yongzhi WANG ; Yu HAN ; Baojin LI ; Yangfan LI ; Yi DU ; Jingjie ZHAO ; Li LI
Chinese Journal of Behavioral Medicine and Brain Science 2015;24(5):407-410
Objective To explore the effect of Chaihu Shugan Powder (CSP) on the content of 5-hydroxytryptamine (5-HT) in the hippocampus and the expression of tryptophan hydroxylase 2(TPH2) in median raphe nuclei in depression rats.Methods SD rats were randomly divided into normal group(n=12),model group (n=12),fluoxetine group (n=12),low-dose and high-dose CSP group (n =12,respectively).Depression model was made by reserpine intraperitoneal injection.During the experiment,the weight and the open-field scores were calculated;the content of 5-HT was detected by ELISA.The expression of TPH2 in median raphe nuclei was detected by Western blot.Results Compared with the weight ((225.02±5.23) g),the open-field scores ((12.6± 5.1)score) and content of 5-HT ((1.09±0.27) ng/ml) in the model group,high-dose CSP showed significantly improve the depressive rats in weight,open field score and content of 5-HT ((238.78±5.16) g,(15.6±7.8) score and (1.80±0.58) ng/ml,respectively;P<0.05 or P<0.01).The expression of TPH2 (0.66±0.21) in median raphe nuclei in the high-dose CSP group was apparently increased compared with that in the model group(0.16±0.04) (P <0.01).Conclusion CSP have the effects of anti-depression,which could be related with the increase of the 5-HT content in the hippocampus and the expression of TPH2 in median raphe nuclei.
4.Inhibitory effect and induction of apoptosis of caffeic acid Ge on growth of U14 in mice
Yueyan HUANG ; Chun XIAO ; Mingjuan WU ; Yangfan YU ; Chunhua LIU ; Youhui XU
Chinese Journal of Pathophysiology 2000;0(08):-
AIM:To evaluate the antitumor effect of caffeic acid Ge on U14 tumor bearing mice.METHODS:The tumor inhibitory ratios of caffeic acid Ge on the growth of U14 in mice was observed.Apoptosis morphological transformation of U14 cells induced by caffeic acid Ge was detected by electronic scan microscope and MG-P staining.Alteration of cell cycle was analyzed by flow cytometry.Apoptosis-related protein levels of Bax and Bcl-2 were determined by immunity histochemistry technology.MTT assay was applied to study the antitumor activities of caffeic acid Ge in U14 cell lines in vitro.RESULTS:Tumor inhibitory rates in caffeic-acid Ge groups were 38.50%,47.17% and 64.02%(from low dose to high dose)(P
5. Decabromodiphenyl ethane: a review of its pollution levels and toxicity
Yangfan YU ; Yi BAI ; Tian CHEN
Chinese Journal of Preventive Medicine 2018;52(8):855-861
Decabromodiphenyl ethane (DBDPE) is a kind of new brominated flame retardants, which is widely used as a replace of decabromodiphenyl ether in electronic appliances, textiles and other goods. This review summarizes environmental levels and body burden of human beings of DBDPE in recent years. The data shows that the concentration of DBDPE in the environment and human tissues shows an upward trend. According to limited experiments about its toxicity, DBDPE shows similar toxicity to decabromodiphenyl ether. DBDPE can interfere thyroid hormones balance, and cause damage to liver, reproductive development, kidney, et al, which implies that DBDPE might be another new persistent organic pollutant. Further researches are needed.
6.Gait abnormalities among elderly persons with type 2 diabetes and peripheral neuropathy
Jiayu ZHU ; Haiyan YU ; Zhenyi WAN ; Yangfan SUN ; Shuai YAO ; Zhida JIANG ; Lan CHEN ; Yu CHEN ; Guilan HUANG ; Rongzheng YUAN
Chinese Journal of Physical Medicine and Rehabilitation 2022;44(12):1090-1094
Objective:To explore the features the gait of elderly persons with type 2 diabetes and peri-pheral neuropathy.Methods:Twenty patients no less than 60 years old with type 2 diabetes and peripheral neuropathy (DPN) formed a DPN group, while 20 counterparts with type 2 diabetes but without peripheral neuropathy composed the DM group, and another 20 healthy counterparts served as a control group. The three groups were tested using the Swedish Qualisys motion capture system and their walking speed, step length, step width, stride frequency and stride length, bipedal foot support phase time, single foot support phase time, peak plantar pressure, and regional-holding time were collected and compared.Results:The average walking speed, stride length and stepping frequency of the DPN group were all significantly lower than the other 2 groups′ averages. Their bipedal support phase was significantly longer, but their single foot support phase time was significantly shorter. And in the DPN group the average first and second peak plantar pressures and the second peak pressure time were significantly greater than the other groups′ averages.Conclusions:Elderly patients with type 2 diabetes and peripheral neuropathy have significant gait abnormalities, decreased walking stability, as well as increased plantar pressure and plantar compression time.
7.Effects of transient receptor potential melastatin 7 on dexamethasone-mediated cytoskeletal protein remodeling of trabecular meshwork cells
Liling LIU ; Jiangang XU ; Zhikun OUYANG ; Yangfan YANG ; Kaili WU ; Minbin YU
Chinese Journal of Experimental Ophthalmology 2018;36(4):259-265
Objective To investigate the effects of transient receptor potential melastatin (TRPM) 7 on dexamethasone (Dex)-mediated cytoskeleton remodeling in human trabecular meshwork.Methods Human trabecular meshwork cells (HTMs) were primarily cultured and the cells of generation 3 to 6 were used in this study.The expression of TRPM7 protein in the cells was located using immunofluorescence technology.Dex at the dose of 0.2 mg was added into culture medium for 4 days with the final concentration of 1×10-5,1×10-6 and 1×10-7 mol/L,respectively.Western blot assay was employed to detect the relative expression level of TRPM7 protein.Cultured cells were divided into non-transfected group,siRNA transfected group,TRPM7-siRNA1 transfected group and TRPM7-siRNA2 transfected group,and the expressions of TRPM7 protein and p-cofilin protein in the cells were assayed by Western blot method.Cultured cells were divided into normal control group,Dex-treated group,siRNA transfected group and TRPM7-siRNA transfected group,and the expression of phalloidin (a cytoskeletal protein) and Vinculin (focal adhesion protein) was detected by immunofluorescence staining.In addition,cultured cells were divided into normal control group,Dex-treated group,2-APB (a Ca2+ inhibitor) treated group,ethylene glycol tetraacetic acid (EGTA) (a calcium chelator)-treated group,TRPM7-siRNA transfected group and TRPM7-siRNA+Dex group,and the [Ca2+] i in the cells was observed by Fluo-3AM immunofluorescence staining.Western blot assay was used to detect the expression of p-cofilin in the cells.Results TRPM7 was positively expressed on the cell membrane.The relative expression of TRPM7 was gradually reduced with an increase of Dex dose (F=4.210,P<0.05),and the expression of TRPM7 was significantly decreased in 1 × 10-5 mol/L Dex group compared with the normal control group (P< 0.05).Western blot assay revealed that the relative expression levels of TRPM7 in the TRPM7-siRNA1 and TRPM7-siRNA2 group were significantly lower than those of non-siRNA transfected group and siRNA transfected group (all at P<0.05).In the Dex-treated group and TRPM7-siRNA transfected group,the cells were enlarged in size with the lessened processes in comparison with the normal control group.Immunofluorescence staining showed that the actin fiber and vinculin increased in the Dex-treated group,and more spread but depolymerized actin fiber was seen in the TRPM7-siRNA transfected group.Compared with the normal control group,the fluorescence intensity of [Ca2×] i was weak in the Dex-treated group and TRPM7-siRNA transfected group.The relative expression levels of p-confilin protein was lower in the TRPM7-siRNA transfected group than that in the siRNA transfected group (0.317 ±0.031 vs.0.092±0.071) (t =5.030,P =0.007).Conclusions Dex induces the downregulation of TRPM7 expression in HTMs.The downregulation of TRPM7 probably participates in Dex-induced cytoskeletal remodeling by causing the depolymerization of actin cytoskeleton and reduction of [Ca2+] i in HTMs.
8.Clinical study of Guben-Tiaoping Decoction in improving the success rate of inhaled corticosteroid stepwise treatment in asthma
Wei GAO ; Weiping WU ; Huiping SU ; Di LUO ; Yangfan LI ; Fang SU ; Minhua CHEN ; Yu LI
International Journal of Traditional Chinese Medicine 2021;43(10):954-959
Objective:To evaluate the effect of combined use of Guben-Tiaoping Decoction on the success rate of stepwise treatment and life quality of asthma patients. Methods:A total of 76 patients with asthma in remission stage who were using ICS were divided into treatment group (38 cases) and control group(38 cases) according to random number table. Both groups were treated with ICS reduction based on the 2017 Gina Protocol. The treatment group was treated with Guben-Tiaoping Decoction for 12 weeks. The number of patients who were able to successfully complete the stepwise treatment after the end of the trial was calculated, the relevant indexes of Asthma Control Test (ACT), TCM Syndrome score, St. George’s respiratory questionnaire (SGRQ) and lung function were collected before and after treatment. Result:After 12 weeks’ treatment, the success rate of stepwise was 89.5% (34/38) in the treatment group and 78.9% (30/38) in the control group ( χ2=14.862, P=0.001). Meanwhile, the ACT score of the treatment group were better than those in the control group (Daytime symptoms t=-3.860, P=0.001, Dyspnea t=-2.007, P=0.40, Night waking t=-2.732, P=0.009, Reliever needed for symptoms t=-2.262, P=0.031, Control situation t=-6.994, P=0.001, Total Score t=-9.562, P=0.001). The score of TCM Syndrome in the treatment group was significantly lower than that of the control group ( P=0.001); PEF in treatment group [(6.92 ± 1.71) L vs. (5.84 ± 1.22) L; t=-2.880, P=0.005] was significantly better than that of the control group. Conclusion:Guben-Tiaoping Decoction is helpful to improve the success rate of ICS stepwise treatment and the life quality of patients.
9.Comparative analysis of constitutes and metabolites for traditional Chinese medicine using IDA and SWATH data acquisition modes on LC-Q-TOF MS
Dian KANG ; Qingqing DING ; Yangfan XU ; Xiaoxi YIN ; Huimin GUO ; Tengjie YU ; He WANG ; Wenshuo XU ; Guangji WANG ; Yan LIANG
Journal of Pharmaceutical Analysis 2020;10(6):588-596
Identification of components and metabolites of traditional Chinese medicines (TCMs) employing liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF MS) techniques with information-dependent acquisition (IDA) approaches is increasingly frequent. A current drawback of IDA-MS is that the complexity of a sample might prevent important compounds from being triggered in IDA settings. Sequential window acquisition of all theoretical fragment-ion spectra (SWATH) is a data-independent acquisition (DIA) method where the instrument deterministically fragments all precursor ions within the predefined m/z range in a systematic and unbiased fashion. Herein, the superiority of SWATH on the detection of TCMs' components was firstly investigated by comparing the detection ef-ficiency of SWATH-MS and IDA-MS data acquisition modes, and sanguisorbin extract was used as a mode TCM. After optimizing the setting parameters of SWATH, rolling collision energy (CE) and variable Q1 isolation windows were found to be more efficient for sanguisorbin identification than the fixed CE and fixed Q1 isolation window. More importantly, the qualitative efficiency of SWATH-MS on sanguisorbins was found significantly higher than that of IDA-MS data acquisition. In IDA mode, 18 kinds of sangui-sorbins were detected in sanguisorbin extract. A total of 47 sanguisorbins were detected when SWATH-MS was used under rolling CE and flexible Q1 isolation window modes. Besides, 26 metabolites of sangui-sorbins were identified in rat plasma, and their metabolic pathways could be deduced as decarbonylation, oxidization, reduction, methylation, and glucuronidation according to their fragmental ions acquired in SWATH-MS mode. Thus, SWATH-MS data acquisition could provide more comprehensive information for the component and metabolite identification for TCMs than IDA-MS.
10.Demographic characteristics of whole blood donors aged 56 to 60 in Guangzhou
Lei WU ; Xuecheng DENG ; Jieying YU ; Yangfan DENG ; Shijie LI
Chinese Journal of Blood Transfusion 2022;35(2):186-188
【Objective】 To study the demographic characteristics of whole blood donors aged 56 to 60 in Guangzhou, so as to provide reference for smooth implementation of the recruitment and management of elderly blood donors in Guangzhou. 【Methods】 The data of whole blood donors, aged 56 to 60, in Guangzhou Blood Center from July 1, 2016 to June 30, 2021 were retrospectively analyzed, and such characteristics as the number of blood donations, gender, donation frequency, donation volume, occupation and educational background were analyzed. 【Results】 A total of 3 996 blood donors aged 56 to 60, including 3 013 males and 983 females, gave 14 416 donations during July 1, 2016 to June 30, 2021, with the proportion of 400 mL specification at 64.1% (6 999/10 918) of males, which was significantly higher than that of females 57.7% (2 017/3 498). The number of blood donors had been decreasing year by year, but the ratio of men to women was stable.The education background of blood donors were mainly high school, junior college, and undergraduate education, accounting for more than 60% in total, and the proportion of undergraduate and graduate degree trended to grow. Staff, workers, medical staff and civil servants were the majorities, and the proportion of medical staff trended to grow. 【Conclusion】 Male blood donors aged 56 to 60 in Guangzhou showed stronger intention to donate blood, and the proportion of blood donors with better education and medical staff was on the rise.