1.EFFECT OF TAURINE ON IRON BIOAVAILABILITY IN RATS AND ITS POSSIBLE MECHANISM
Meizi YANG ; Yongjun JIN ; Shuping ZHANG ; Weidong LIU
Acta Nutrimenta Sinica 1956;0(04):-
Objective To investigate the effect of taurine on iron bioavailability in rats and its possible mechanism. Method SD rats were randomly divided into three groups:normal group (NG),taurine treated group (TG),experimental control group (EG). TG and EG were induced to be anemic experimentally for 4w before the study. Then the rats were fed iron supplements for 25 d,meanwhile TG was treated with taurine following hemoglobin(Hb) repletion bioassay. Regression analysis of TG and AG were measured following Hb repletion bioassay using iron supplements. And the relative biological value of TG following Hb repletion were calculated. The serum iron,unsaturated iron-binding capacity(UIBC),total iron-binding capacity(TIBC),transferin saturation(TS)and the expression of hepcidin were detected. Results The iron bioavailability of TG was higher than EG,and the RBV of TG was 126%. Other hematological variables of TG were close to NG,and better than EG. The expression of hepcidin was increased compared with NG,but decreased compared with EG. Conclusion Taurine can improve iron bioavailability in rats via suppressing hepcidin expression in rats.
2.Identification of daturae flos and its adulterants based on DNA barcoding technique.
Jianping HAN ; Meini LI ; Kun LUO ; Meizi LIU ; Xiaochen CHEN ; Shilin CHEN
Acta Pharmaceutica Sinica 2011;46(11):1408-12
To identify the original plant of Daturae Flos from its adulterants by DNA barcoding, the sequences of ITS2, psbA-trnH, matK, rbcL of four species including Datura metel, Darura innoxia, Darura stramonium and Brugmansia arborea were compared and analyzed. The PCR and sequencing success rate of the four regions (ITS2, psbA-trnH, matK, rbcL) was 100%, 90%, 100% and 85%, respectively. Sequences were assembled with CodonCode Aligner. K2P distances were calculated and NJ tree was performed by MEGA 4.1. Thirty SNPs were found among ITS2 sequences, and 33 insert/deletes were found among psbA-trnH intergenic regions. The interspecific K2P distance of ITS2 and psbA-trnH was obviously higher than that of the intraspecific one. As to matK and rbcL, there was no "Barcoding Gap" existing between inter- and intra-specific distances. The NJ trees of the four regions/combinations were built separately. Samples of Brugmansia arborea were clustered into one clade, and the other species of Datura L. formed another clade. The results showed that either ITS2 or psbA-trnH was useful to identify Daturae Flos from its adulterants.
3.Effect of agomelatine on memory impairment and ERK5 expression in hippocampus of rats with posttraumatic stress disorder
Meizi WANG ; Chaomeng LIU ; Guiqing ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2018;27(11):971-975
Objective To investigate the effect of agomelatine on memory impairment and expres-sion of extracellular signal-regulated kinase 5 (ERK5) in post-traumatic stress disorder ( PTSD)-like rats. Methods Forty-eight SD rats were divided into control group (SHAM group),post-traumatic stress disorder group (PTSD group),agomelatine group (AGO group) and placebo control group(PC group) according to random number table with 12 in each group. The PTSD-like model was established by internationally recog-nized single prolonged stress (SPS) stimulation,and the AGO group and PC group were given the same a-mount of agomelatine and saline respectively by intragastric administration within 8 hours after modeling for 14 days. The arousal emotional level and learning and memory ability of rats were observed by open field ex-periment and Morris water maze test. Then the expressions of ERK5 and ERK5 mRNA in hippocampus of rats were detected by Western blot and qRT-PCR. Results (1) The results of the open field experiment showed that compared with the SHAM group (38. 58±5. 76),the numbers of crossing the grids of the PTSD group (29. 75±3. 75) decreased (P<0. 01). Compared with the PC group (28. 58±2. 91),the number of crossing the grids of the AGO group (41. 00±4. 49) increased (P<0. 01). (2) In Morris water maze test, positioning navigation experiment showed that compared with the SHAM group, the escape latency of the PTSD group and the PC group increased (P<0. 01),while the escape latency of AGO group was shorter than that of the PC group (P<0. 01). And in the space exploration experiment,compared with the SHAM group (2. 12±0. 51),the times of crossing the platform in PTSD group (1. 03±0. 43) and the PC group (1. 23± 0. 59) decreased (P<0. 01),while the times of crossing the platform in AGO group (2. 75±0. 72) increased compared with PC group (P<0. 01). Compared with SHAM group (12. 14±2. 53),the latency of crossing the platform of PTSD group (27. 33±6. 54) increased (P<0. 01),and the agomelatine group (14. 36±4. 27) de-creased compared with the PC group (29. 67±9. 72) (P<0. 01). (3) Results of Western blot and qRT-PCR showed compared with the SHAM group,the levels of ERK5 and ERK5 mRNA in rat hippocampus were up-regulated in the PTSD group (P<0. 01),and the expression of ERK5 and ERK5 mRNA was higher than that of PC group (P<0. 01). Conclusion Agomelatine can effectively improve the learning and memory ability of PTSD-like rats,which may be related to the up-regulation of ERK5 protein expression in hippocampal tis-sues.
4.Effect of oxytocin on behavior and MKP-1 expression in hippocampus of rats with posttraumatic stress disorder
Wenlong XING ; Meizi WANG ; Ruohuan SHI ; Chaomeng LIU ; Guiqing ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2018;27(3):230-234
Objective To investigate the effect of oxytocin on the expression of MKP-1 level in the hippocampus of post-traumatic stress disorder rats.Methods Eighteen SD rats were randomly divided into the control group,administration group and the experimental group with 6 rats in each group.The administration group and the experimental group were treated with internationally recognized single prolongation stress (SPS) method to stimulate the rats in order to establish PTSD models.The 14-day oxytocin intervention was given to rats of the administration group after the SPS stimulation within 8 hours.And the behavioral changes of rats were observed by open-field test and Morris water maze test.The changes of MKP-1 mRNA in the hippocampus of rats were detected by real-time quantitative PCR (qRT-PCR),and the levels of MKP-1 Protein in the hippocampus of rats were detected by Western Blot.Results (1) Compared with the model group (2.50± 1.05 and 22.16±7.14),the times of the standing and crossing grid section quantities in the open-field test in the administration group(5.16± 1.17and 32.83±5.71) and control group (6.67±2.16 and 39.83± 4.62) significantly decreased (P<0.05).Morris water maze showed that the incubation of the model group was markedly prolonged (P<0.05) compared with that of the administration group,while in spatial probe test,the incubation period of the rats in administration group was prolonged,and the number of wearing stage decreased (P<0.05).(2) Compared with the administration group (1.30±0.03),the expression of MKP-1 mRNA in hippocampus of rats in model group (4.04±0.46) was notably up-regulated (P<0.05).And the protein level of MKP-1 in model group(1.95±0.68) was also increased compared with that in administration group (1.46±0.27) (P<0.05).Conclusions Oxytocin can protect the learning and memory ability and reduce the stress-related performance of rats via regulating the expression of MKP-1.
5.Correlation between event-related potential P300 and violent behavior in patients with schizophrenia
Danyu LI ; Chaomeng LIU ; Meizi WANG ; Lan XIAO ; Guiqing ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2021;30(8):722-727
Objective:To explore the change characteristics of event-related potential P300 in different violence risk levels of schizophrenic patients and analyze the risk factors of violence in schizophrenic patients.Methods:Totally 158 schizophrenic patients in Lyuzhou hospital of Shihezi City from January 2019 to August 2020 were collected and assessed with the violence risk scale for 3 days.According to the assessment results, the patients who met the inclusion criteria were divided into low-risk group( n=78), medium-risk group( n=51) and high-risk group( n=29). The auditory P300 of patients in each group was completed within 3 days and act of violence was observed and recorded within one week.Data analysis was carried out by SPSS 20.0 software.The changes of P300 in different violence risk groups were analyzed by ANOVA, and the influencing factors of violence in patients with schizophrenia were analyzed by logistic regression. Results:There was no significant difference in latency of P300 among the three groups (χ 2=4.71, P=0.10), but there was significant difference in amplitude of P300( F=6.67, P<0.01). Compared with the low-risk group ((12.14±9.19) μV), the amplitude of P300 in medium-risk group ((8.25±7.13) μV) and high risk group ((6.71±4.97) μV) decreased significantly ( t=-3.14, -5.45, both P<0.05). There was no significant difference in amplitude of P300 between the high-risk group and the middle-risk group( t=-2.31, P>0.05). The latency and amplitude of schizophrenia patients with violent behavior were significantly different from those without violent behavior ( Z=-6.30, 9.78, both P<0.01). High BVC grade (compared with high-risk group, low-risk group: OR=0.03, 95% CI : 0.00-0.35; the middle risk group: OR=0.09, 95% CI : 0.01-0.62), prolonged P300 latency ( OR=1.30, 95% CI : 1.13-1.48) and decreased P300 amplitude ( OR=0.50, 95% CI: 0.36-0.70), delusion of victimization ( OR=0.12, 95% CI: 0.02-0.76)were the risk factors for violent behavior. Conclusion:The latency and amplitude of P300 can be used as the reliable neuroelectrophysiological indicators for evaluating violence risk in patients with schizophrenia.It has important clinical application value for evaluating violence in patients with schizophrenia.
6.Quantitative analysis of macular structure and microvascular changes in patients with diabetic macular ischemia
Changying LIU ; Lingna LI ; Meizi GAO ; Xuzheng ZHAO
Chinese Journal of Ocular Fundus Diseases 2024;40(2):116-121
Objective:To observe the changes of macular structure and microvessels in eyes with diabetes macular ischemia (DMI).Methods:A retrospective case study. From January 2023 to July 2023, 23 patients of 31 eyes diagnosed with DMI at Tangshan Ophthalmological Hospital were included in this study. Among them, there were 14 males with 23 eyes; Female cases with 8 eyes. Age were (59.5±4.6) years old. According to the DMI grading standard formulated by the research group for early treatment of diabetes retinopathy, the patients were divided into mild DMI group, moderate DMI group, and severe DMI group, with 8, 12, and 11 eyes respectively. The blood flow density (VD), perfusion area (FA), small vessel VD (SVD), inner retinal capillary plexus VD, FA, and outer retinal, choroidal, and ganglion cell complex (GCC) thickness within 1 mm of the macular fovea in retinal superficial vascular plexus (SVP)were measured using a scanning frequency light source optical coherence tomography instrument. The changes in macular structure and microvasculature in the affected eyes of different degrees of DMI groups were compared and observed. Inter group comparisons were conducted using one-way ANOVA or Kruskal Wallis H-test. Spearman correlation analysis was used to analyze the correlation between DMI severity and GCC, outer retina, choroid thickness, VD, FA and SVP VD, SVD and FA in inner retina. Results:The GCC ( F=70.670), outer retinal thickness ( H=12.393), VD ( F=105.506), SVD ( H=25.300), FA ( F=107.655), and VD ( H=24.098) and FA ( H=25.300) of the retinal SVP in the mild, moderate, and severe DMI groups were compared, and the differences were statistically significant ( P<0.05). There was no statistically significant difference in choroidal thickness ( H=2.441, P>0.05). Pairwise comparison between groups: VD, SVD, FA of GCC thickness and SVP, and VD of inner retina were statistically significant between severe DMI group and moderate DMI group, and between moderate DMI group and mild DMI group ( P<0.05). The thickness of outer retina was statistically significant between severe DMI group and moderate DMI group ( P<0.05). Inner retinal FA: there were statistically significant differences between severe DMI group, moderate DMI group and mild DMI group ( P<0.05). The correlation analysis results showed that GCC ( r s=-0.918), outer retinal thickness ( r s=-0.448), and inner retinal VD ( r s=-0.894) and FA ( r s=-0.918), as well as VD ( r s=-0.919), SVD ( r s=-0.924), and FA ( r s=-0.939) of retinal SVP, were all negatively correlated with the degree of DMI ( P<0.05). There was no correlation between choroidal thickness and degree of DMI ( r s=-0.081, P>0.05). Conclusion:The thickness of GCC, outer retina and choroid, the VD, SVD, and FA of the retinal SVP, the VD and FA of inner retina are all reduced in eyes with different degrees of DMI, while all of them are negatively correlated with the degree of DMI, except for choroid thickness.
7.The inhibitory effect of FoxF2 shRNA on the expression of extracellular matrix of human trabecular meshwork
Aihua LIU ; Meizi GAO ; Liangyu HUANG ; Xun LIU ; Ruihong SU ; Jinzhi ZHAO ; Liming WANG ; Xiaomin ZHANG ; Xiaorong LI ; Lijie DONG
Chinese Journal of Experimental Ophthalmology 2019;37(6):405-410
Objective To explore the role of forkhead box F2 (FoxF2) in the extracellular matrix of trabecular meshwork.Methods The cultured human trabecular meshwork cells (HTMCs) were divided into Scramble control group and FoxF2 small hairpin RNA (shRNA) group,then FoxF2 shRNA,the FoxF2 restructuring interference carrier was built,HTMCs were infected with FoxF2 shRNA lentivirus.Western blot assay was used to detect the expression of FoxF2 protein and extracellular matrix.Furthermore,Transwell counting experiment was used to analyze the migration ability of HTMCs.Results The cultured HTMCs grew well and showed a long spindle shape.The growth status of HTMCs was well,and their morphological characteristics were consistent with the HTMCs in vivo.The relative expression level of FoxF2 protein in the FoxF2 shRNA group was lower than that in the Scramble control group,with a significant difference between them (0.72 ± 0.02 vs.1.27 ± 0.05;t =16.68,P < 0.01).The relative expression level of fibronectin (FN),collagen type Ⅰ (COL Ⅰ) and α-smooth muscle actin (α-SMA) were 0.43±0.03,0.53 ±0.08 and O.86±0.15 in the FoxF2 shRNA group,and 0.87±0.04,1.66±0.06 and 1.73 ±0.13 in the Scramble control group,respectively,the relative expression levels of FN,COL Ⅰ and α-SMA in the FoxF2 shRNA group were significantly lower than those in the Scramble control group (t =15.08,18.81,7.50,all at P<0.01).The migration number of HTMCs in the FoxF2 shRNA group was significantly lower than that in the Scramble control group (117.30±11.41 vs.251.00±10.37;t =8.72,P<0.01).Conclusions The FoxF2 shRNA lentivirus are successfully constructed,which can decrease the expression of FoxF2 in HTMCs.Low expression of FoxF2 can reduce the expression level of extracellular matrix protein in HTMCs and inhibit the migration ability of HTMCs.
8.Regulation of Krüppel-like factor 6 via activating transcription factor 4 pathway to apoptosis of human lens epithelial cells
Fang TIAN ; Jinzhi ZHAO ; He TENG ; Liangyu HUANG ; Xun LIU ; Ruihong SU ; Meizi GAO ; Xiaomin ZHANG ; Xiaorong LI ; Lijie DONG ; Hong ZHANG
Chinese Journal of Experimental Ophthalmology 2018;36(3):181-186
Objective To investigate the regulating effects of Krüppel-like factor 6 (KLF6) on the apoptosis of human lens epithelial cells (HLECs) by activating transcription factor 4 (ATF4) pathway and explore the bio-molecular mechanism of KLF6/ATF4-induced HLECs apoptosis.Methods HLECs (HLE-B3) were cultured using high glucose DMEM medium.The eukaryotic expression plasmid pEGFP-C2-ATF4 was transfected into the cells by liposome 2000 in the ATF4-transfected group,and pEGFP-C2 was transfected in the empty plasmid group.Then the cells were exposed to 20 mJ/cm2 ultraviolet ray B (UVB) for 200 seconds,The morphological changes of the cells were observed by hematoxylin & eosin staining and Hoechst33258 fluorescein staining.Cultured cells were transfected using pEGFP-C2-KLF6 and pEGFP-C2 plasmid and pSilencer-KLF6 (siKLF6) and pSilencer plasmid,respectively,and the expression of ATF4 protein in the cells was detected by Western blot assay.Culture cells were divided into four groups.pEGFP-C2 and pSilencer plasmids were co-transfected into the cells in the empty plasmid group;pEGFP-C2-KLF6 and pSilencer empty plasmid were co-transfected into the cells of the KLF6 + pSilencer group;pEGFP-C2 empty plasmid and pSilencer-ATF4 were co-transfected in the cells of the siATF4 + pEGFP-C2 group;pEGFP-C2-KLF6 and pSilencer-ATF4 plasmids were co-transfected in the cells of the KLF6 + siATF4 group,and then the cells were exposed to UVB.The apoptosis of the cells were detected by ELISA assay.Results Cultured cells grew well in the normal control group with the uniform morphology and regular arrangement.The karyopyknosis,karyorrhexis and enlargement of intercellular space were found in the cells exposed to UVB.In the ATF4 transfected group,the number of cells was decreased.The relative expression level of the ATF4 protein in the cells was 0.99±0.06 and 0.13±0.02 in the UVB+ATF4 transfected group and UVB+pEGFP-C2 plasmid group,respectively,with a significant difference between them (t =23.13,P<0.01).The relative expression levels of KLF6 and ATF4 proteins in the KLF6 transfected group were higher than those in the empty plasmid group,and the relative expression levels of KLF6 and ATF4 proteins in the siKLF6 group were significantly lower than those in the empty plasmid group (all at P<0.01).ELISA assay showed that the apoptotic rate in the ATF4 transfected group was 1.37± 0.11,which was significantly higher than 0.31 ±0.11 in the normal control group (t =8.034,P =0.001);the apoptotic rate of the cells was increased in the KLF6+pSilencer group and decreased in the siATF4+pEGFP-C2 group in comparison with the empty plasmid group (P<0.01,P=0.02).In addition,the apoptotic rate in the KLF6+ siATF4 group was remarkably lower than that in the KLF6 + pSilencer group (P< 0.01).Conclusions KLF6 promotes the apoptosis of HLECs induced by UVB radiation.Silence of ATF4 gene reduces the apoptotic rate of the cells.ATF4 is probably a target factor in the regulating oathwav of KLF6 to apoptosis.
9.Effect of dl-3-n-Butylphthalide on apoptosis of retinal müller cells induced by hydrogen peroxide
Xiaoli XING ; Liangyu HUANG ; Ruihong SU ; Xun LIU ; Jinzhi ZHAO ; Meizi GAO ; Xiaomin ZHANG ; Xiaorong LI ; Lijie DONG
Chinese Journal of Ocular Fundus Diseases 2018;34(5):481-486
Objective To observe the protective effect of dl-3-n-Butylphthalide (NBP) on apoptosis of retinal Müller cells induced by hydrogen peroxide (H2O2).Methods Human retinal Müller cells cultured in vitro were divided into normal control group,model group (H2O2 group) and experimental group (H2O2+NBP group).The cells in the H2O2 group and H2O2+NBP group were cultured with 200 μ mol/L H2O2 for 2 h.Then the culture solution of the H2O2 group replace with complete medium and the H2O2+NBP group replace with complete medium containing 1 tmol/L NBP.The normal control group was a conventional cultured cells.Müller cells were identified by immunofluorescence staining.Hematoxylin-eosin (HE) staining was used to observe the apoptosis morphological changes.MTT assay was used to detect the activity of of retinal Müller cells after after 24 h and 48 h of NBP intervention.Hoechst33258 staining was used to observe the apoptosis.LIVE/DEAD (R)cell activity/cytotoxicity kit was used to detect cell viability.Dichlorofluorescein diacetate (DCFH-DA) + endoplasmic reticulum (ER) red fluorescent probe (ER-Tracker Red) double staining was used to observe the expression level of reactive oxygen species (ROS) in ER of cells.One-way ANOVA combined with Dunnett statistical method were used for data analysis.Results HE staining showed that the number of cells in H2O2+NBP group was higher than that in H2O2 group.MTT assay showed that after 24 h and 48 h of NBP intervention,the differences in cell viability between the normal control group and the H2O2 group,the H2O2 group and the H2O2+NBP group were statistically significant (t=28.96,3.658,47.58,20.33;P<0.001,0.022).The results of Hoechst33258 showed that the nuclear nucleus of a few cells in the H2O2+NBP group was crescent-shaped and the nuclear fragmentation was reduced,and the blue fluorescence of the remaining cells was uniform.The LIVE/DEAD ~ cell activity/cytotoxicity kit showed that the number of dead cells with red fluorescence in the H2O2 group increased significantly,and the number of viable cells with green fluorescence decreased significantly.In the H2O2+NBP group,the number of viable cells with green fluorescence increased,and the number of dead cells with red fluorescence decreased.The double staining results of DCFH-DA+ER-Tracker Red showed that the green fluorescence intensity of H2O2 group was significantly enhanced;the green fluorescence intensity of H2O2+NBP group was lower than that of H2O2 group.Conclusion NBP alleviates H2O2-induced apoptosis of human retinal Müller cells by inhibiting ROS production.