1.Dynamic expression and mechanism of γδ T cells in the spleen of mouse with experimental autoimmune uveitis
Beibei, WANG ; Qingmei, TIAN ; Xiaofeng, XIE ; Jike, SONG ; Hongsheng, BI
Chinese Journal of Experimental Ophthalmology 2017;35(9):793-798
Background Previous studies showed that the pathogenesis of uveitis is related to γδ T cells.However,it remains unclear that how these cells are involved in experimental autoimmune uveitis (EAU).Objective This study aimed to observe the dynamic changes of γδ T cells in EAU and explore the role of γδ T cells in the pathological process of EAU.Methods Forty-five C57BL/6(B6) mice were assigned to the normal control group (six mice) and EAU model group (thirty-nine mice).The mice were immunized subcutaneously at 6 spots on the footpads,tail base,and flank with emulsion containing human interphotoreceptor retinoid binding protein1-20 (IRBP1-20) emulsified in complete Freund's adjuvant.After immunization,the mice were examined for clinical signs of EAU by using a Genesis-D camera.The changes of histopathology were compared by hematoxylin and eosin staining.Mouse lymphocytes were isolated and purified from the spleens of IRBP1-20-immunized or normal B6 mice by using a γδ T-cell isolation kit.Flow cytometry was used to detect the changes of intracellular expression of interleukin-17A (IL-17A),and then transferred the activated γδ T cells into EAU models to analyze the changes of clinical signs and histopathology of EAU.Experimental study program as well as the use and feeding of the animals were authorized by the Animal Management and Use Committee of Shandong Traditional Chinese Medicine University.Results The inflammatory symptoms in mouse eyes appeared on day 12 after modeling.The initial changes were fundal blood vessel thickening and minimal inflammatory cell infiltration.Then,multifocal chorioretinal lesions,serious vasculitis and linear lesions were observed on days 16-20,along with abundant lymphocyte infiltration in the vitreous and retinal disorganization.The inflammation symptom scores and the pathological inflammation scores at different time points after modeling had statistically significant differences (F =51.399,P =0.000;F =47.342,P =0.000).The inflammatory symptoms in the eyes began to abate from day 28 onwards.The number of γδ T cells was obviously increased during the inflammation phase of EAU at day 16-20 after modeling,with the number of γδ T cells was (5.67 ±-0.49) % and (5.78 ±±0.55) %,respectively,which was significantly higher than (1.53 ± 0.14) % before modeling,with significant differences between them (both at P<0.05),meanwhile CD69 levels and the integrin lymphocyte function-associated antigen-1 (LFA-1) and secreted IL-17A were elavated.The secretion level of IL-17A was (13.40±0.50)% and (17.80±2.37)% on day 16 and day 20 after modeling,respectively,which was significantly higher than (1.53 ± 0.19) % before modeling,with significant differences between them (P =0.000,0.001).The activated γδ T cells were transferred into EAU model,the inflammation symptom scores were 1.00 (1.00,2.00) after activated γδ T cells were transferred into EAU model,which was significantly higher than 0.75 (0.05,1.00) of the untransferred group (Z =27.00,P =0.03),and the symptoms of EAU were aggravated.Conclusions The proportion of γδ T cells reaches peak in inflammation of EAU,and the cells are activated.The activated γδ T cells in the EAU model play a immune regulation role by secreting IL-17A.
2.The dynamic changes of regulatory T cells in the pathogenesis of experimental autoimmune uveitis
Lian, ZHANG ; Jike, SONG ; Kai, TANG ; Fengming, ZHENG ; Junguo, GUO ; Hongsheng, BI
Chinese Journal of Experimental Ophthalmology 2016;34(8):684-690
Background Studies show that regulatory T cells (Treg) are a kind of T cell subsets to negatively regulate immune response,and play an important role in maintaining immune homeostasis and immune tolerance.Autoimmune uveitis is an autoimmune disease,the regulation of Treg cells in pathogenesis and progression of autoimmune uveitis is not fully unelucidated.Objective This study was to observe the dynamic changes of Treg in experimental autoimmune uveitis (EAU) rats and explore the role of Treg cells in the pathological process of EAUrats.Methods Eighty four 6-8 week-old SPF Lewis rats were randomly divided into model group and control group.The mixed emulsifier of interphotoreceptor retinoid-binding protein (IRBP)1177-1191,tuberculin (TB),complete Freund adjuvant (CFA) and PBS (300 μl) was subcutaneously injected in double rear foot pad,abdominal side and back,and only equal amount of TB,CFA and PBS emulsifier was used in the same way in the control group.Ocular inflammation symptoms was examined at 9,13,18,23,28,35 and 48 days after modeling and scored based on the severity of the inflammatory.Six rats of each group were sacrificed in above time points respectively for the histopathological examination of iris,ciliary body and retinas by haematoxylin-eosin staining.The lymphocytes were isolated and cultured from rat spleens,and the proportion of Foxp3-labelled cells,a specific marker of Treg cells,was assayed by flow cytometry.The relative expression level of Foxp3 mRNA in the lymphocytes detected by using realtime quantitative PCR (RT-PCR).The use and care of the rats complied with the ARVO Statement.Results Eye inflammatory response appeared at 8 days after immunization,showing vasodilation and hyperemia of rat iris in the model group,and the response peaked at 13 days,with exudation and hypopyon in the anterior chamber.The highest inflammatory scores were 3.75±0.42 at day 13,and the ocular inflammation reaction was gradually relieved after that and disappeared at 23 days after immunity.A significant difference in ocular inflammatory scores of model rats was found among different time points (F =81.709,P < 0.001);while no inflammatory symptom was observed in the control group.Histopathology examination showed obvious infiltration of inflammatory cells in the iris,ciliary body and retinas in model rats,including neutrophils,lymphocytes and mononuclear cells.The proportion of Foxp3-labelled cells in spleen lymphocytes was (5.50 ± 0.64)%,(13.36 ± 0.98)%,(10.34 ± 0.79)%,(9.58 ± 1.02)%,(6.73 ±0.81)% and (5.58 ± 0.47) % in the model group on day 13,18,23,28,35,48 respectively,with statistically significant differences in comparison with (2.80 ± 0.38) %,(3.36 ± 0.53) %,(3.65 ± 0.57) %,(3.37 ± 0.43) %,(3.33±0.50)% and (3.13±0.61)% in the control group (t=-6.272,-15.556,-11.910,-9.753,-6.154,-5.491,all at P<0.01).The change trend of Foxp3 mRNA expression was consistent to the dynamic change of the proportion of Foxp3-labelled cells.Conclusions The pathogenesis and development is closely associated with the dynamic changes of CD4+ CD25 + Foxp3+ Treg cells in EAU rats.
3.Analysis of 114 cases of warfarin-related major bleeding in the emergency department
Yuanli LEI ; Yunchao NI ; Shouquan CHEN ; Peisen ZHOU ; Junyan CHENG ; Jike XUE ; Wenxing SONG ; Zhangping LI
Chinese Journal of Emergency Medicine 2021;30(3):336-341
Objective:To investigate the anticoagulant causes, hemorrhagic susceptibility factors and clinical characteristics of patients with warfarin-related major bleeding in the emergency department of a general hospital.Methods:In a registry study from January 2017 to February 2020, 114 cases of warfarin-related major bleeding patients admitted to Department of Emergency Medicine, the First Affiliated Hospital of Wenzhou Medical University were enrolled. The descriptive methods were used to analyze anticoagulant causes, hemorrhagic susceptibility factors and clinical characteristics. Patients were divided into the international normalized ratio (INR) overrange group and the INR non-overrange group according to INR value during bleeding. The Chi-square test, Student’s t test, and Wilcoxon rank sum test were used to compare the differences between the INR overrange group and the INR non-overrange group. The Wilcoxon rank sum test and Student’s t test were used to analyze the characteristics of gastrointestinal tract bleeding and cerebral hemorrhage. Results:Warfarin-related major bleeding accounted for 0.36% (114/32 040) of first aid cases and 9.84% (114/1 158) of warfarin-taking cases, respectively. Seventy-seven cases (67.5%) of anticoagulant causes were related to atrial fibrillation (AF) and 32 cases (28.1%) were related to post-operative cardiac valve replacement. Of the bleeding susceptibility factors, HAS-BLED scored at 4.0 (3.0, 5.0), 84 cases (73.7%) had a history of drug use, 77 cases (67.5%) aged older than 65 years old, 65 cases (57.0%) had irregular INR monitoring, and 29 cases (25.4%) had recent increase in dose. Forty cases (35.1%) were gastrointestinal tract bleeding with the lowest hemoglobin (Hb) value and the highest score of HAS-BLED. Twenty-one cases (18.4%) were cerebral hemorrhage with the shortest prothrombin time (PT), the lowest INR value, the highest Hb, and the lowest score of HAS-BLED. Twelve cases (10.5%) died or gave up treatment in critical condition, including 6 cases of cerebral hemorrhage, 5 cases of gastrointestinal tract bleeding, and 1 case of hemoptysis. There were statistically significant differences in previous history of antiplatelet therapy, recent increase in dose, HAS-BLED score and bleeding site between the INR overrange group and the INR non-overrange group (all P<0.05). Conclusions:Among patients with warfarin-related major bleeding, AF and post-operative cardiac valve replacement are the main causes of warfarin anticoagulation. INR overrange is related to the previous history of antiplatelet therapy, recent increase in dose, and the high score of HAS-BLED. The gastrointestinal tract bleeding is the most common, with the lowest Hb value and the highest score of HAS-BLED. Cerebral hemorrhage is the second common, with the shortest PT, the lowest INR value, and the highest Hb. The incidence and mortality rates of warfarin-related major bleeding are relatively high.
4.Follow-up study of factors influencing prognosis of cardiac arrest patients after acute myocardial infarction
Yuanli LEI ; Wenxing SONG ; Jinmei LU ; Yijin HE ; Zhangping LI ; Jike XUE ; Aiwen HE ; Qianqian ZHANG ; Jincun SHI ; Yingru LU ; Shouquan CHEN
Chinese Journal of Emergency Medicine 2018;27(7):740-746
Objective To assess the factors associated with the restoration of spontaneous circulation (ROSC) and 2-year survival prognosis in patients with cardiac arrest (CA) after acute myocardial infarction (AMI),and after ROSC,the effects of various factors on midian survival time and on 2-year survival.Methods In a registry study from January 2005 to January 2015,all consecutive AMI-induced CA patients treated with cardiopulmonary resuscitation (CPR) admitted to our hospital were enrolled.The survivors were followed-up for 2 years.Univariate analysis was applied to evaluate factors associated with rate of ROSC and 2-year survival.Multivariable logistic regression analysis was applied to evaluate statistically significant factors in the univariate analysis.Medians with inter-quartile ranges were used to describe 2-year survival time affected by various factors after ROSC.Kaplan-Meier survival curve analysis was used to evaluate the effect of factors on 2-year survival.Results A total of 254 cases with CA after AMI were enrolled,including 129 cases of ROSC and 71 cases of 2-year survival.Univariate analysis showed age ≥ 70 years,CA occurred during 22:00-8:00,the duration time ofCPR ≥ 15 min and adrenaline dosage > 5 mg were unfavorable predictors of ROSC;while,left ventricular ejection fraction (LVEF) ≥ 40% before CA,shockable rhythm and percutaneous coronary intervention (PCI) therapy were favorable predictors.Besides,age ≥ 70 years,intubation during CPR,adrenaline dosage > 5 mg and cardiogenic shock were unfavorable predictors of 2-year survival;While,male,normal daily activity before CA and PCI treatment were favorable predictors.Multivariable analysis showed age,the duration of CPR,adrenaline dosage,LVEF before CA,the rhythm during CPR and PCI therapy were independent predictors of ROSC.Age and PCI therapy were independent predictors of 2-year survival.Among patients,the survival time was affected by various factors after ROSC,and the factors with minimum 25% and small median value were associated with cardiac rupture,cancer,adrenaline dosage > 5 mg and cardiogenic shock.The factor with maximum 25% value was PCI treatment (216 days).Kaplan-Meier survival analysis suggested that age ≥ 70 years was an unfavorable factor of 2-years survival (Log-rank test,P=0.007);while,PCI treatment was a favorable factor (Log-rank test,P<0.01).PCI-related prognosis analysis showed that the effectiveness of PCI was related to the timing of PCI,the number of infarctrelated artery and the difference in culprit lesion.Conclusions The age ≥ 70 years was disadvantageous to both ROSC and 2-year survival.PCI treatment was favorable to both ROSC and 2-year survival.
5.Research updates on surgical treatments for portal hvpertension
Jinwei YANG ; Zhen MA ; Jike HU ; Tianliang SONG ; Xiaohong LIU ; Chunyu GENG ; Zhijian HAN ; Yumin LI ; Hao CHEN
Chinese Journal of Hepatobiliary Surgery 2017;23(9):640-645
Portal hypertension is a common clinical syndrome in chronic liver disease,such as schistosomiasis,portal vein occlusion cirrhosis and so on,which can be diagnosed when the hepatic venous pressure gradient is (HVPG) > 5 mmHg (1 mmHg =0.133 kPa).It could lead to gastroesophageal varicose veins rupture,ascites,spontaneous bacterial peritonitis,hepatorenal syndrome,hepatopulmonary syndrome,hepatic encephalopathy and some other serious complications,and is the primary death cause in cirrhosis and liver transplantation.The development of portal hypertension has experienced 4 phases ineluding the research about portal hypertension related theories and animal trial phase,preclinical tests and data accumulation phase,devascularization and shunts rapid development phase,the development phase of new technologies such as interventional and endoscopic surgical treatment,liver transplantation since the middle of the 19th century.The surgical procedures have been modified,which greatly reduce the complication and improve the life quality after operation.But so far none of them can cure portal hypertension thoroughly.This paper not only introduces the pathophysiologic basis of the surgical treatment,but also reviews the history of its development to summarize the recent progress,which may facilitate its surgical treatment.
6.Progress in the application of swept-source optical coherence tomography in corneal imaging
Lili ZHAO ; Jike SONG ; Xiuzhen LU ; Hongsheng BI
International Eye Science 2024;24(7):1058-1063
Advances in imaging technology have revolutionized the field of ophthalmology, changing the understanding, diagnosis, and treatment of ophthalmic diseases. Swept-source optical coherence tomography(SS-OCT)is a non-contact high-resolution imaging technology. It further improves imaging depth and scanning speed, adds new algorithms and features. The application of SS-OCT enables the three-dimensional evaluation of corneal structures, offering curvature and height maps for both the anterior and posterior surfaces of the cornea, as well as precise corneal thickness mapping. These invaluable tools aid ophthalmologists in effectively screening and diagnosing various corneal lesions such as keratoconus, corneal dystrophy, and degeneration. Moreover, the enhanced speed, accuracy, and sensitivity provided by SS-OCT measurements facilitate improved surgical planning and postoperative monitoring for patients undergoing refractive surgery or keratoplasty. This article reviews the development of SS-OCT technology and its potential clinical utility in corneal diseases and surgical application, in order to support more possible future research and clinical treatment.
7. Establishment and practice on collaborative education model of "medicine, teaching, research and prevention" in Optometry
Hongying TANG ; Lian ZHANG ; Wenjun JIANG ; Jike SONG ; Hongsheng BI
Chinese Journal of Medical Education Research 2019;18(9):902-906
In order to satisfy the social and medical demands for optometric talents, the education model combining "medicine, teaching, research and prevention" was established, fully integrated with various resources like talents, facilities, equipment from the affiliated eye hospital, institute of ophthalmology and prevention and treatment center for juvenile myopia. Cultivation of high-quality applicative talents which integrated "knowledge, skills and accomplishment" were taken as the main line, Cultivation goal was established in accordance with requirement of talent. The curriculum system was optimized on the basis of competence. The teaching mode was reformed to "2+2" mode, "double-position teachers" became the requirement for the teaching staff and professional education comprehensive practice platform of "one entity, two wings" was constructed, with good effectiveness being achieved.
8.Comparison of effects of different fixation methods on the posterior pole of guinea pig eyeballs
Xiuyan ZHANG ; Yixue YIN ; Jike SONG ; Hongsheng BI
International Eye Science 2024;24(8):1207-1212
AIM: To avoid the problem of retinal dissection in guinea pig large eyeball tissue sections, different methods were used to optimize the fixation effect of the posterior pole of the eyeball.METHODS: A total of 75 normal guinea pigs(2 weeks old)were randomly divided into 5 large groups. Group A(1-3 small groups), the entire eyeball was fixed with FAS, Davidson fixative 1(D1), and Davidson fixative 2(D2)for 24 h; group B(4-6 small groups), the entire eyeball was fixed with FAS, D1, and D2 for 1 h, then cut the cornea and fix it in their respective fixatives for 2 h; group C(7-9 small groups), the eyeball was fixed in FAS, D1, and D2 for 1 h, divided into left and right halves along the direction of the optic nerve, and then placed them in their respective fixation solutions for 2 h; group D(10-12 small groups), after fixation for 3 h in FAS, D1, and D2, the eyeball was divided into left and right halves along the optic nerve direction; group E(13-15 small groups), the cornea was cut after fixation for 3 h in FAS, D1, and D2. Hematoxylin-eosin(HE)staining was used to compare the fixation effect on posterior eyeball in each group.RESULTS: After fixation, the surface of the eyeballs in groups, 1-6 and 11-15 was smooth and round, with a transparent and bright color. In groups 7-10, the eyeballs were sunken, wrinkled, and deformed. The HE staining showed that the eyeball morphology of groups 1, 5, 6, 14, and 15 was significantly better than the other groups, with a regular internal tissue structure. The eyeballs of the other groups were sunken and wrinkled, and the internal tissue was curled and tangled, with severe retinal detachment. In groups 1, 5, 6, 14, and 15, the retina, choroid, and sclera tissues of group 14 were closely connected, without obvious retinal detachment, rupture, or curling. The tissue structure was clear and visible, and the cells were arranged neatly.CONCLUSION: The fixation effect of cutting the cornea after fixing guinea pig eyeball with D1 fixative for 3 h is the most ideal, and this operation method is simple and suitable for studying the related structures of the posterior pole of the eye.