1.Effect and safety of different concentrations of disintegrin Echistatin to early posterior capsule opacification in diabetic rabbits
Guangxia, QIAN ; Shaojian, TAN ; Hao, LIANG ; Yingying, CHEN
Chinese Journal of Experimental Ophthalmology 2013;32(11):975-979
Background Posterior capsular opacification (PCO) is a primary cause of blurred vision after extra-capsular cataract extraction (ECCE),and there is a higher incidence of PCO in the patients with diabetes mellitus.Echistatin (Ecs) can suppress the proliferation of lens epithelial cells (LECs) and thereby inhibit the formation of PCO.However,its mechanism and safe dose deserve to study.Objective The aim of this study was to observe the inhibitory effect of different concentrations of Ecs on LECs proliferation in the early stage of PCO in diabetic rabbits and explore the safe dose of Ecs.Methods Diabetic mellitus was induced by injection of 90 mg/kg alloxan via ear vein in 15 New Zealand white rabbis.ECCE was performed in the right eyes of the rabbits.The rabbits were randomized to the control group and 5.0,7.5,10.0 and 15.0 μg/ml Ecs group according to randomized number table method.Ecs of 0.2 ml in above doses was injected into the anterior chamber after ECCE in different concentrations of Ecs groups,and 0.2 ml distilled water was used in the same way in the only diabetic rabbits as the control group.Postopeartive response of ocular anterior segment was observed and PCO was graded under the slit lamp microscope.The corneal and retinal specimens were prepared 10 days after operation for the assay of preliferative cell nuclear antigen (PCNA) expression in LECs by immunochemistry to evaluate the effective dose of Ecs.Regular histopathological examination was performed,and apoptosis of corneal endothelial cells and retinal cells was detected by TUNEL method to assess the safe concentration of Ecs.Results Different degrees of corneal edema and exudation in anterior chamber were seen in the eyes of different groups.The inflammatory response disappeared 3-5 days in the control group and 5.0 μg/ml Ecs group and 7 days in the ≥7.5 μg/ml Ecs groups.PCO was 1-2 grade in the control group and 5.0 μg/ml Ecs group and 0 grade in the ≥ 7.5 μg/ml Ecs groups.The difference in the positive expression level (absorbance,A) for PCNA in LECs was significantly different among the control group and various Ecs groups (F=18.006,P=0.001),and the positive expression level of PCNA in the ≥ 7.5 μg/ml Ecs groups was markedly reduced in comparison with that in the control group (P =0.010,0.001).Hematoxylin and eosin staining showed an normal morphology and order arrangement in corneal endothelial cells and intact structure in retinal internal limiting membrane in the groups.TUNEL assay revealed that the apoptosis values (mean A value) of corneal endothelial ceils and retinal cells in the ≤ 10.0 μg/ml Ecs groups were not significantly changed in comparison with the control group (all at P>0.05),but the apoptosis values in the 15.0 μg/ml Ecs group were markedly higher than those in the control group (P=0.004,0.018).Conclusions Ecs can inhibit the early PCO in diabetic rabbits and show the optimal effect at the concentrations of 7.5 and 10.0 μg/ml without visible eytotoxicity to eye other tissues.Therefore,these two doses of Ees might be used for the study of long-term therapeutic effectiveness.
2.Rehabilitation improves the cognition and activity of daily living of patients with vascular dementia
Riguang HU ; Peijun SUN ; Xianxiu LIU ; Jianbo BI ; Guangxia LIANG ; Liying WANG
Chinese Journal of General Practitioners 2010;9(2):126-127
Sixty-fourth patients with vascular dementia were recruited in the study and randomly assigned in rehabilitation (medication plus rehabilitation)and control group (medication alone).The treatment period lasted for 12 weeks,the scorings of mini mental statement examination(MMSE) and activity of daily living scale (ADL) were compared before and 4,8,12 weeks after treatment in both group.The scorings of MMSE and ADL in rehabilitation group and control group were both improved after treatment(P <0.05),but the scorings in rehabilitation group were better than those in control group(P<0.05).
3.A preliminary study on ultrasonic classification and its clinical value of adhesive intestinal obstruction
Liang LI ; Guangxia WANG ; Yunfeng CUI
Chinese Journal of Ultrasonography 2020;29(8):690-694
Objective:To establish the ultrasonic classification criteria of adhesive intestinal obstruction (AIO) and to discuss its clinical value.Methods:The data of 124 cases of AIO confirmed by surgery and pathology in Tianjin Nankai Hospital from January 2014 to October 2019 were retrospectively analyzed. The ultrasonic image features of AIO were summarized. According to the severity of ultrasound images it can be divided into three different types: simple (mild) AIO, complex (moderate) AIO and critical (severe) AIO. The surgery plan was simulated according to the ultrasonic classification results and compared with the actual surgery results.Results:①The ultrasonic classification results of 124 cases of AIO were as follows: simple (mild) AIO in 73 cases (58.9%), complex (moderate) AIO in 16 cases (12.9%), critical (severe) AIO in 35 cases (28.2%). ②The characteristic ultrasonic images of simple (mild) AIO were adhesion of the intestinal wall to the intestinal wall and the intestinal wall to the abdominal wall; the characteristic ultrasonic images of complex (moderate) AIO were adhesive band and adhesive mass; the characteristic ultrasonic images of critical (severe) AIO were internal hernia, volvulus, intussusception, intestinal ischemia and necrosis, and bloody ascites. ③According to the results of ultrasound classification, 124 cases of AIO were treated with simulated surgical procedures, and the coincidence rate with the surgical results was high (87.9%).Conclusions:US classification of AIO is helpful for clinical evaluation of the severity of AIO and can provide imaging basis for clinicians to make the correct treatment plan.
4.Sulforaphane Suppressed Stemness Characters of Lung Adenocarcinoma Stem Cells by Down-regulating Methylation of miR-200c Promoter
Guangxia LIANG ; Xituan QIN ; Wei XIE ; Youke XIE
Cancer Research on Prevention and Treatment 2021;48(10):934-940
Objective To investigate the effect of sulforaphane (SFN) on the proliferation and self-renewal of lung cancer stem cells and its regulatory mechanism. Methods MTT method was used to detect the effect of SFN on the proliferation of lung adenocarcinoma cell lines H460 and A549; tumor sphere formation experiment was used to detect the ability of tumor sphere formation; Western blot was applied to explore the expression of stemness-related proteins (such as β-catenin, Klf4, c-myc) in lung adenocarcinoma cells before and after SFN treatment; NGS sequencing was used to analyze the effect of SFN on the expression profile of tumor cell miRNAs. qRT-PCR verified the changes in the transcription level of key miRNAs by SFN. Western blot was used to detect the effect of SFN on the expression of DNMTs in tumor cells. We constructed miR-200c promoter-GFP plasmid, and applied IF, methylation PCR and DNA sequencing methods to detect the effect of SFN on the methylation level of tumor spheres and miRNA promoter. Results The miRNAs expression profile of lung adenocarcinoma tumor spheres changed significantly after SFN (5.0μmol/L) treatment, and miRNA-200c increased the most. Compared with the control group, the expression of β-catenin, Klf4, c-myc and Vimentin genes in H460 and A549 cells of SFN-S group decreased, and the protein expression levels of DNMT1 and DNMT3a were also significantly decreased. Compared with the control group, H460 and A549 cells stably expressing pEGFP-R200c plasmid in SFN-S group significantly reduced tumor sphere diameter, while tumor sphere fluorescence intensity increased, and GFP protein expression was up-regulated. There were 9 CpG-rich regions in the miR-200c promoter region in the above-mentioned pEGFP-R200c plasmid cell line, and the methylation levels were 88.9%, 44.4% and 38.8% in the control group, SFN-S group and 5-Aza-dC group, respectively. Conclusion SFN may downregulate the expression of stem-related genes in lung cancer stem cells by epigenetically decreasing the methylation level of miR-200c promoter and promoting the transcription of miR-200c.
5.Treatment of cough variant asthma based on the pathogenic characteristics of "wind phlegm and blood stasis"
Zheng GUO ; Yiming HOU ; Yuan LIANG ; Xiuying SI ; Guangxia PAN ; Qihui HU ; Youpeng WANG
International Journal of Traditional Chinese Medicine 2022;44(7):721-724
The wind, phlegm, and blood stasis are important pathogenic factors of cough variant asthma in children, and they are also the pathological products in the occurrence and development of this disease. They have typical pathogenic characteristics. The main pathogenesis characteristics of cough caused by wind, phlegm and blood stasis are as follows that external wind attacks the lungs and induces internal wind, phlegm and dampness accumulates in the lungs, and the lungs fail to declare and descend, and blood stasis obstructs the collaterals and stagnation of Qi. The wind, phlegm, and blood stasis have their own pathogenic characteristics, and their cough-causing also have their own pathogenic characteristics and clinical characteristics. Based on the characteristics of wind, phlegm, and blood stasis, the application of medicine based on the differentiation of symptoms and signs can effectively prevent and treat this disease, and provide theoretical basis and treatment ideas for the treatment of cough variant asthma in children with Chinese medicine.
6.Effects of liraglutide on myocardial autophagy and lipid deposition in early diabetic rats
Guangxia XI ; Ping AN ; Dengyao LIANG ; Yu FENG ; Xuemei FAN ; Kun YANG ; Wei REN
Chinese Journal of Endocrine Surgery 2021;15(5):531-535
Objective:To investigate the effects of liraglutide on the expressions of autophagy markers LC3B, LC3B mRNA and lipid deposition in myocardial tissue of rats with early diabetes mellitus.Methods:A total of 36 healthy male Wistar rats aged 4-5 weeks and weighing (l80-200) g were selected and divided into normal group (group NC, 10 rats) and model group (26 rats) according to random number table. Rats in the NC group were fed with routine diet and rats in the model group were given high glucose and high fat diet for 12 weeks. Rats in the model group were injected with streptozotocin into the abdominal cavity in a single dose of 25 mg/kg after molding. Rats in the model group were further divided into three groups: T2DM group (group DM/NS, 9 rats, given equal volume of saline) , liraglutide intervention group (group DM/LIR, 8 rats, injected with 100 μg/kg liraglutide twice daily) and Liraglutide and Chloroquine intervention group (group DM/LIR+CQ, 8 rats, injected with 100 μg/kg liraglutide twice daily, and injected with Chloroquine 50 mg/kg once every two days) . Rats in group NC were given equal volume of saline. At the end of 12 weeks, all the rats were tested blood glucose and anaesthetized to collect myocardial tissues to observe myocardial lipid deposition and fiber arrangement under light microscope after HE staining. The expressions of LC3B were detected by immunohistochemical method, and the expressions of LC3B mRNA were detected by real-time fluorescence quantitative polymerase chain reaction method. Differences among groups were compared by one-way analysis of variance, pairwise comparison was conducted by using LSD-t test, correlation was analyzed by Pearson correlation.Results:(1) Compared with group NC, the myocardial fibers arranged in disorder, and the ratio of myocardial foam cells/total cardiomyocytes were increased, the level of LC3B mRNA and LC3B were decreased in group DM/NS and DM/LIR+CQ (in group DM/NS: 2.18±0.90 vs 11.79±0.74, 2.03±0.10 vs 1.85±0.06, 194.18±10.19 vs 175.99±6.09, t=25.24, 4.69, 3.22, respectively; in group DM/LIR+CQ: 2.18±0.90 vs 11.24±1.29, 2.03±0.10 vs 1.89±0.08, 194.18±10.19 vs 176.73±7.82, t=17.56, 4.65, 3.99, respectively, all P<0.05) . There is no difference in above indicators (2.18±0.90 vs 1.29±0.60, 2.03±0.10 vs 2.01±0.20, 194.18±10.19 vs 201.27±11.35, t=2.20, 0.28, 1.40, respectively, all P>0.05) . (2) Compared with group DM/NS, the ratio of myocardial foam cells/total cardiomyocytes, the level of LC3B mRNA and LC3B were no difference in group DM/LIR+CQ ( t=1.09, 1.18,0.22, respectively, all P>0.05) . The ratio of myocardial foam cells/total cardiomyocytes was decreased, the level of LC3B mRNA and LC3B were increased in group DM/LIR (11.79±0.74 vs 1.29±0.60, 1.85±0.06 vs 2.01±0.20, 175.99±6.09 vs 201.27±11.35, t=31.86, 2.39, 5.82, respectively, all P<0.05) . (3) The significant negative correlation were observed between the ratio of myocardial foam cells/total cardiomyocytes and LC3B mRNA, LC3B levels (r=-0.977, -0.986, respectively, all P<0.05) . Conclusion:Liraglutide can protect the myocardial structure in early diabetic rats by increasing myocardial autophagy, reducing the number of myocardial foam cells, and improving the myocardial lipid deposition.