1.Protective effect of Bu Shen Yang Xue Ming Mu Formula on oxidative injury in ARPE-19 cells
Qiang, CHEN ; Na, AN ; Zeng-Yuan ZHUANG ; Li-Na, LIANG
International Eye Science 2017;17(8):1433-1436
AIM: To explore the effect of Bu Shen Yang Xue Ming Mu (BSYXMM) Formula on hydroquinone-induced oxidative stress injury in ARPE-19 cells.METHODS: The oxidative injury model of ARPE-19 cell was induced by exposure to various concentrations of hydroquinone (HQ) to determine the optimal concentration.Intestinal absorption solutions of BSYXMM Formula were prepared.Effect of intestinal absorption solutions of BSYXMM Formula on the cell viability was detected by CCK-8 assay,and the percentage of apoptotic cells was measured by TUNEL assay.The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in ARPE-19 cells were detected by means of chemical colorimetry.RESULTS: It was found that ARPE-19 cell viability significantly decreased when the concentration of HQ was higher than 90μmol/L.Compared with the model group,1% and 2% intestinal absorption solutions in the pre-treatment group could significantly alleviate HQ-induced injury (P<0.01) and 0.5% and 5% intestinal absorption solutions in the pre-treatment group could alleviate the injury in certain degree(P<0.05).While in the treatment group 1% and 2% intestinal absorption solutions could alleviate the injury to some extent (P<0.05).TUNEL results showed that the apoptosis rate decreased significantly in the pre-treatment group (P<0.01)and to some extent in the treatment group (P<0.05)compared with the model group.It was shown that both levels of SOD and GSH-Px in pre-treatment group and treatment group were markedly higher than that of model group(P<0.05),and pre-treatment group had more significant effect (P<0.01,P<0.05).CONCLUSION: BSYXMM Formula could protect against HQ-induced oxidative stress injury in ARPE-19 cells,which may be related with the increasing of antioxidant enzyme in the cells.
2.Effects of the prescription of reinforcing kidney, nourishing blood, improving eyesight on ARPE - 19 cells induced by acrolein
Man, LI ; Li-Na, LIANG ; Zeng-Yuan, ZHUANG
International Eye Science 2015;(5):777-780
?AlM: To explore the effects of the prescription of reinforcing kidney, nourishing blood, improving eyesight on the oxidative stress model of ARPE-19 cells induced by acrolein.
?METHODS:SD rats serum containing the prescription of reinforcing kidney, nourishing blood, improving eyesight and the content of distilled water in serum were prepared. The effects of the prescription and distilled water in serum at different concentration ( 2. 5%, 5%, 10%, 20% and 40%) on cell vitality was observed by cell counting kit ( CCK-8 ) assay. the logarithmic phase of ARPE-19 cells were pretreated by different concentrations (1. 25%, 2. 5%and 5%) of the prescription serum and distilled water in serum for 24h. Then it was treated with 75μmol/L acrolein for 24h. Cell vitality was observed by CCK-8 assay. The change of cell nucleus was detected by DAPl staining .
?RESULTS: 2. 5% and 5% serum had no effect on cell viability (P>0. 05), while 10%, 20%, 40% serum could inhibit cell viability (P<0. 05). CCK-8 results showed that 2. 5% and 5% the prescription of reinforcing kidney, nourishing blood, improving eyesight serum was better than distilled water in serum (P<0. 05).
?CONCLUSlON: The prescription of reinforcing kidney, nourishing blood, improving eyesight has the protective effect on ARPE-19 cell damage induced by acrolein.
3.Measurement and influencing factors on retinal vascular caliber
Man, LI ; Yong-Sheng, YANG ; Na, AN ; Zeng-Yuan, ZHUANG
International Eye Science 2014;(9):1608-1611
Retinal vessel is the only visible vessels in human body and systemic microvascular lesions could be evaluated by the degree of retinal vessels. ln this article, the progress of retinal vascular caliber measuring method and its influencing factors were briefly reviewed.
4.Suppression of p38 mitogen-activated protein kinases inhibitor on myofibroblasts transdifferentiation and extracellular matrix synthesis after filtration surgery in rabbit eyes
Kun, ZENG ; Li-na, HUANG ; Fang-wei, YING
Chinese Journal of Experimental Ophthalmology 2012;30(8):729-733
Background The main cause of filtering surgery failure is over proliferation of fibroblasts in filtering channels,leading to excessive fibrosis and scar formation.Researches determined that p38 mitogen-activated protein kinase(MAPK) signaling pathway plays an important role in fibroblast phenotype transition. Objective The present study was to investigate the inhibitory effect of p38 MAPK inhibitor on myofibroblasts transdifferentiation and the extracellular matrix synthesis after filtration surgery in rabbit eyes. Methods Trabeculectomy was performed on 24 eyes of 12 clean New Zealand white rabbits to establish the filtering operative models.The models were randomized into model group,SB203580 group and mitomycin C ( MMC ) group.1 ml SB203580 ( 0.2 g/L) was conjunctively injected at the end of operation in the rabbits of the SB203580 group,and the cotton piece with 0.2 g/L MMC solution was placed on the operative area for 3 minutes intraoperatively in the rabbits of the MMC group.The bleb appearances were examined under the slit lamp microscope,and intraocular pressure(IOP) was measured with Icare tonometer I,3,7,10,14 days after operation.0.2 ml aqueous humor was extracted and the conjunctive tissue at the filtering area was obtained 14 days after operation for the detection of α-smooth muscle actin (α-SMA) and fibronectin protein by ELISA.Expression of ACTA2 mRNA,connective tissue growth factor(CTGF) mRNA and alpha2 chain of type Ⅰ collagen( COL1A2 )mRNA in conjunctive tissue was assayed with fluorescence real-time PCR. Results Vascularization of fibrosis of filtering bleb were obvious in the eyes of the model group,and the bleb was flat and diffuse in the eyes of the SB203580 group and MMC group on 14 days following operation.No significant difference was seen in IOP before trabeculectomy among these three groups( F=0.065,P=0.937 ).IOP was gradually elevated with the increase of time after operation ( F =32.873,P =0.030 ).ELISA assay showed that α-SMA level in conjunctiva was lower in the SB203580 group and MMC group compared with the model group,and that of MMC group was significant lower than the SB203580 group( P<0.05 ).Fibronectin level in conjunctiva was lower in the SB203580 group and MMC group compared with the model group,and that of MMC group was significant lower than the SB203580 group (P<0.05).Fluorescence real-time PCR showed that expressions of the ACTA2 mRNA,CTGF mRNA and COL1A2 mRNA were significantly different among the three groups( P<0.01 ),with the highest expression in model group and the lowest expression in the MMC group ( P < 0.05 ). Conclusions Fibrotic reaction after trabeculectomy can be suppressed by inhibiting p38 MAPK signal pathway.The mechanism of SB203580 is to reduce the synthesis of myofibroblasts transdiffercntiation and extracellular matrix.
5.Design and practice of clinical case-based learning in medical imaging teaching
Na DUAN ; Jing LI ; Liang ZENG ; Zhongqiu WANG
Chinese Journal of Medical Education Research 2021;20(1):32-35
Objective:To explore the application value of clinical case-based learning in medical imaging teaching.Methods:A total of 145 clinical medical students from three classes in the third year were enrolled and they were divided into three classes. One class took case-based learning, one class took traditional teaching, and one class took both. The teaching content was the preclinical medical professional course—Medical Imaging. Continuous variables were expressed in accordance with normal distribution (mean standard deviation). Differences in mean and median values were evaluated by using t test and the Mann-Whitney U test, respectively. Categorical data were compared using the chi-square test. Results:There was a significant difference between the combined teaching class and the case-based learning class or the traditional teaching class in the final exam ( P=0.002). There was also a significant difference between the combined teaching class and the traditional teaching class in the film reading quiz ( P=0.035). More than 70% of students recognized the case-based learning. Conclusion:Case-based learning is helpful to improve students' flexible application of relevant knowledge and skills of analysis and communication.
6.Protein Interaction Network Construction and Biological Pathway Analysis Related to Atherosclerosis.
Quhuan LI ; Shanshan GU ; Na LI ; Zhenyang LI ; Wenlong LAI ; Yang ZENG
Journal of Biomedical Engineering 2015;32(6):1255-1260
Atherosclerosis is a complex disease characterized by lipid accumulation in the vascular wall and influenced by multiple genetic and environmental factors. To understand the mechanisms of molecular regulation related to atherosclerosis better, a protein interaction network was constructed in the present study. Genes were collected in nucleotide database and interactions were downloaded from Biomolecular Object Network Database (BOND). The interactional data were imported into the software Cytoscape to construct the interaction network, and then the degree characteristics of the network were analyzed for Hub proteins. Statistical significance pathways and diseases were figured out by inputting Hub proteins to KOBAS2. 0. The complete pathway network related to atherosclerosis was constructed. The results identified a series of key genes related to atherosclerosis, which would be the potential promising drug targets for effective prevention.
Atherosclerosis
;
genetics
;
Databases, Factual
;
Humans
;
Protein Interaction Mapping
;
methods
;
Protein Interaction Maps
;
Software
7.Quantitative measurement of macular ganglion cell complex thickness by spectral domain optical coherence tomography
Ning, FAN ; Li-na, HUANG ; Jing, HE ; Xiao-li, SHEN ; Kun, ZENG ; Hong-bo, CHENG
Chinese Journal of Experimental Ophthalmology 2012;30(8):743-747
Background Glaucoma is primarily characterized by the damage of retinal ganglion cells.The macular ganglion cell complex (GCC)thickness can be quantitatively measured using spectral domain optical coherence tomography(SD-OCT). Objective This clinical study was to explore the macular GCC thickness change in primary open-angle glaucoma (POAG) patient with SD-OCT. Methods A serial case-controlled study was designed.A total 101 eyes of 101 POAG patients and 41 normal eyes of 41 age- and refract power-matched normal subjects were cnrolled in the study.POAG patients were assigned to normal perimetry POAG group,early stage POAG group,advanced POAG group and late stage POAG group.Average macular GCC thickness(GCC-Avg),superior GCC thickness(GCC-Sup) and inferior GCC thickness (GCC-Inf)of subjects were measured by SD-OCT and compared among POAG patients and normal controls.Peripapillary retinal nerve fiber layer(RNFL) thickness was measured with time domain OCT(TD-OCT).The correlation between GCC thickness with RNFL thickness or mean deviation(MD) of perimetry were evaluated and analyzed.Informed consent was obtained from each patient prior to entering this study.Results GCC-Avg thickness,GCC-Sup thickness and GCC-Inf thickness were significantly decreased in the normalperimetry POAG group and early stage POAG group compared with the normal control group (GCC-Avg:t =5.411,10.247,P < 0.01 ; GCC-Sup:t =6.171,9.484,P< 0.01 ; GCC-Inf:t =5.281,8.592,P < 0.01 ).Also,GCC-Avg thickness,GCC-Sup thickness and GCC-Inf thickness were significantly decreased in the advanced POAG group compared with the early stage POAG group ( GCC-Avg:t =4.246,P<0.01 ; GCC-Sup:t - 2.419,P - 0.019 ; GCC-Inf:t =4.636,P<0.01 ),and GCC-Avg thickness,GCC-Sup thickness and GCC-Inf thickness were significantly decreased in the late stage POAG group compared with the advanced POAG group (GCC-Avg:t=2.095,P=0.040;GCC-Sup:t=2.756,P<0.01:GCC-Inf:t =2.018,P =0.040 ).The positive correlations were seen between GCC-Avg thickness,GCC-Sup thickness,GCC-Inf thickness and RNFL-Avg thickness,RNFL-Sup thickness,RNFL-Inf thickness respectively( r =0.802,0.825,0.856,P < 0.01 ).MD value of perimetry was positive correlated with GCC-Avg thickness in POAG patients ( r =0.601,P < 0.01 ). Conclusions SD-OCT can quantitatively measure and differentiate the GCC thickness in POAG patients.The GCC thickness gradually decreases with the development of POAG.There exist a well correlation between visual field defect and RNFL thinning.
8.Preliminary study of functions of ompW gene in Salmonella paratyphi A
Qin CHEN ; Na LI ; Haoyu LIANG ; Bin WANG ; Hua WEI ; Ming ZENG
Chinese Journal of Microbiology and Immunology 2012;32(1):6-11
ObjectiveTo construct ompW- and ompW+ mutants of Salmonella paratyphi A with λRed system,and then study the function of the gene preliminarily.Methods Homologous regions were amplified from the genome Salmonella paratyphi A 50973,and then connect with kana fragment from plasmid pET22b-kan to construct a recombinant vector.The resultant fragments were amplified and transferred into 50973 with the help of λRed system after its concentration.Then the ompW- mutant was obtained PCR identification.Connect the recombinase expression plasmid pACU184 with full fragment of ompW regulatory region and coding region,then transfer the connection product into the mutant,the ompW+ mutant was obtained after double digest identification.Full cells of the wild,ompW- and ompW+ mutants were samples for SDS-PAGE and Western blot to detect the expression of protein OmpW.Biochemical identification of wild strain and mutant strains was conducted,so did the growth curves of the wild and the ompW- mutant.Choose BALB/c mice as a model to determine median lethal dose LD50 of the wild and mutant strains in order to observe the correlation between ompW gene and bacterial virulence.ResultsompW gene was knocked out in Salmonella paratyphi A 50973,also the ompW+ mutant was constructed; The wild and ompW+ mutant express the protein OmpW,while the ompW- mutant lost the protein.Each of the wild and mutant strains was Salmonella paratyphi A,and no obvious difference could be observed for their growth curves.LD50 for each strain was also similar.Conclusion The ompW gene has no correlation with the virulence in S.paratyphi A 50973,but the contribution of the mutants made an important foundation for the further study of functions of the gene ompW in Salmonella paratyphi.
9.Imbalance between regulatory T cells and Th 17 cells in patients with systemic sclerosis
Xinjuan LIU ; Na GAO ; Mengtao LI ; Henghui ZHANG ; Yong HOU ; Dong XU ; Xiaofeng ZENG
Chinese Journal of Rheumatology 2016;20(3):148-153
Objective To investigate the levels, phenotypes and functions of regulatory T cells (Treg) and Th17 cells in systemic sclerosis (SSc) patients and assess the role of imbalance between Treg and Th17 cells in the pathogenesis of SSc. Methods Peripheral blood mononuclear cells (PBMCs) of 31 SSc patients and 33 healthy controls were analyzed for the expression of CD4, CD25, CD45RA, the cytotoxic T-lymphocyte antigen-4 (CTLA-4), FoxP3, and IL-17 using flow cytometry. Treg immunosuppression capacities were measured in co-cultured experiments. The expressions of IL-17A, IL-22, IL-6 and IL-1β were measured by FlowcytoMix. The expression of FoxP3, CTLA-4, IL-17A, and RORC mRNA were measured by real-time polymerase chain reaction (PCR). The independent samples t-test was used to compare data between the groups. Results The frequency of Treg cells was significantly elevated in SSc [(3.6 ±1.1)%] compared with controls [(2.0±0.8)%, t=6.88, P<0.01)] and the expression of CTLA-4 was lower in Treg cells of SSc (P=0.034). The expression of CTLA-4 and FoxP3 mRNA was lower in SSc compared with controls (P<0.05), the immunosuppressive capacity of Treg cells were diminished in SSc (P=0.034). Th17 cells were increased in SSc (P<0.01); the levels of IL-17A, IL-22, IL-6 and IL-1β in the serums of SSc were higher than the controls (P<0.05). In SSc, CD25+FoxP3+IL-17+cells were increased[0.075%±0.032)%vs (0.049±0.027)%, t=3.52, P=0.029], but no difference of CD25+FoxP3+IL-17- was observed when compared with control. The expression of IL-17A and RORC mRNA were increased in SSc (P<0.01, respectively). Conclusion Both Treg and Th17 cells are increased in patients of SSc. Treg increases along with the deficiency of phenotype and function. FoxP3+IL-17+co-express cells are increased, and the balance between Treg and Th17 cells is broken.
10.Observation of the efficacy and safety of the Song-Relaxing and Zhen-Vibrating abdomen manipulation for patients with prediabetes
Min LI ; Na PENG ; Jiaofei ZENG ; Ying ZHOU ; Yihui DENG ; Qinghu HE ; Xiehe KONG
Journal of Acupuncture and Tuina Science 2022;20(1):58-64
Objective: To observe the safety of the Song-Relaxing and Zhen-Vibrating abdomen manipulation in treating patients with prediabetes and its effects on blood glucose and lipid metabolism.Methods: One hundred and two patients with prediabetes were divided into a manipulation group and a control group according to the random number table method, with 51 cases in each group. All patients received the general behavioral intervention for prediabetes, with additional Song-Relaxing and Zhen-Vibrating abdomen manipulation in the manipulation group and oral metformin hydrochloride tablets in the control group. Both groups received the intervention for six months. Results: Fourteen patients dropped out during the treatment, and a total of 88 patients completed the trial, including 45 cases in the manipulation group and 43 cases in the control group. After the treatment, the prediabetes control rate was 93.3% in the manipulation group and 74.4% in the control group, and the difference between the two groups was statistically significant (P<0.05); no patient in the manipulation group progressed to diabetes, while the rate of conversion to diabetes in the control group was 6.9%, with a significant difference between the two groups (P<0.05). After the treatment, the body mass index (BMI), glycosylated hemoglobin A1c (HbA1c), fasting plasma glucose (FPG), 2-hour postload plasma glucose (2hPG) level during an oral glucose tolerance test (OGTT), fasting insulin (FINS), homeostasis model assessment-2 of insulin resistance (HOMA2-IR), total cholesterol (TC), and triglycerides (TG) decreased in both groups versus baseline, with significant differences within the groups; the levels of all indicators were lower in the manipulation group than in the control group (P<0.05), and the differences between the two groups were more prominent at the sixth month (P<0.01). At the sixth month, the high-density lipoprotein cholesterol (HDL-C) in the manipulation group was increased, while there was no significant change in the control group. In the control group, three patients reported mild gastrointestinal reactions at the initial dosing, which improved after medication adjustment. No other adverse events were observed in either group. Conclusion: Both metformin hydrochloride tablets and the Song-Relaxing and Zhen-Vibrating abdomen manipulation can improve blood glucose and lipid metabolism and reduce insulin resistance and clinical discomfort in patients with prediabetes, but the Song-Relaxing and Zhen-Vibrating abdomen manipulation has higher efficacy and safety.