1.Expression of interleukin-17 and retinoid-related orphan nuclear receptor gamma t in cornea of experimental keratomycosis
Zong-sheng, ZENG ; Xiao-li, HAN ; Jian-zhang, HU
Chinese Journal of Experimental Ophthalmology 2013;(7):653-658
Background In the past few decades,the balance of Th1/Th2 is often used to explain the immune mechanisms of fungal infection and fungal disease.More recently,a novel subset of CD4+ effector Th cells has been found to participate in anti-fungal infection response.However,whether Th17 is involved in the immune response in fungal keratitis is unclear up to now.Objective Present study was to investigate the expression change of Th17 type cytokine and its specific transcription factor,retinoid-related orphan nuclear receptor gamma t (RORγt),in the cornea of Fusarium solani keratitis.Methods Ninety-six clean BALB/c mice were divided into Fusarium solani keratitis model group and control group by randomized digital table.Fusarium solani keratitis models were established by epikeratophakia-assisted corneal epithelial erasion and interlayerly injection of 5 μl (1 × 106 CFU/ml) Fusarium solani solution in the right eyes,and the equal volume of PBS was injected in the same way in the control group.10% KOH wet film was used to examine the fungal hyphea and funga strain was identified by inoculation.The corneas were examined under the slit lamp microscope 1 day,3,5,7 days after modeling and the inflammatory response was scored based on the criteria of Wu and Hu.The histopathological examination of corneas was performed in the time points above.Real time fluorescence quantitative PCR was used to detect the expression levels of interleukin-17 (IL-17) mRNA and RORγt mRNA in the corneas.The expression of IL-17 protein in the corneas was detected by ELISA.The use and raise of the mice followed the Statement of Association for Research in Vision and Ophthalmology.Results The inflammatory scores were 3.2±0.8,6.6± 1.1,9.4± 1.1 and 6.8±0.8 in 1 day,3,5,7 days after modeling,showing a significant difference among them (F =89.786,P =0.010).The inflammatory scores were higher in the third and seventh day than that in the first day (P<0.05),but they were significantly lower than that in the fifth day (P<0.05).The infiltration of inflammatory cells showed a coincident tendency with the score.The expressing levels of IL-17 mRNA (2-ΔΔCt) in the corneas were 4.12±0.73,20.72±1.81 and 14.16±1.88 in 3,5,7 days after modeling,with statistically significant differences in comparison with those in the control group (P<0.01),and the expression level was significantly higher in the fifth day than those in the first,third and seventh day in the model group(P<0.01).The expression levels of IL-17 protein (ng/g) were significantly increased 1 day,3,5,7 days in the model group compared with the control group (P<0.01).A similar change was found in the expression of RORγt mRNA to that of IL-17 mRNA.Conclusions Expressions of IL-17 and its transcription factor RORγt upregulate in the fungal keratitis and has an association with inflammatory degree,which suggests that Th17 subset may play an important role in the immune responses of fungal keratitis.
2.Research and advancement of RNA.
Xiao-han XIAN ; Jin-song HU ; Zhang-min YANG ; Yi-li WANG ; Lü-sheng SI
Chinese Journal of Pathology 2003;32(3):276-278
3.Research of ITK regulation on mouse spleen lymphocytes proliferation and differentiation.
Zong-Hui XIAO ; Feng HE ; Hai-Lan YAO ; Ji-Sheng HAN ; Zhe-Wei LIU
Chinese Journal of Experimental and Clinical Virology 2009;23(4):269-271
OBJECTIVEBy using the RNAi method to inhibit Itk protein expression specificity, to observe lymphocytes proliferation and cytokines production, verify its function as a drug target.
METHODSDesigned siRNA aims at Itk sequence according to its sequence and solid structure, then electrotransfected into mouse spleen lymphocytes, We validated the decrease of Itk protein by Western-Blot, and detected the change of the cell proliferation by MTS and the change of inflammatory cytokines by ELISA.
RESULTSItk protein can be suppressed by Itk-siRNA, there were significantly reduced compared to its control group on cell proliferation as well as cytokine secretion such as IL-2, IL-4, IL-5, IFN-gamma. They all have statistical difference (P < 0.05).
CONCLUSIONItk has an important immunomodulatory effect in mouse spleen lymphocytes proliferation and secretion of inflammatory cytokines.This can supply an experimental basis to regard Itk as drug target for inflammation therapy.
Animals ; Cell Differentiation ; Cell Proliferation ; Cytokines ; genetics ; immunology ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Protein-Tyrosine Kinases ; genetics ; immunology ; Spleen ; cytology ; immunology
4.Application of rapid immunohistochemical staining technique in intraoperative frozen section diagnosis of thyroid neoplasm
Xi-lin LIU ; Ping-ling YANG ; Yong-sheng ZHANG ; Jie HAN ; Xiao-ling ZHU ; Ping GAO
Cancer Research and Clinic 2011;23(12):810-813
ObjectiveTo investigate the application of rapid immunohistochemical staining technique in intraoperative frozen section diagnosis of thyroid neoplasm.Methods MaxVision one-step rapid immunohistochemical staining technique was used to detect the expression of CK19,HBME-1,and Gal-3 in frozen section of papillary thyroid carcinoma(PTC)andthyroid benign lesions.MaxVision conventional immunohistochemistry of frozen remaining tissue was served as control.ResultsMaxVision one-step rapid immunohistochemical staining technique could be completed in 20 minutes.The positive localizations of three markers detected by rapid immunohistochemistry were similar to conventional immunohistochemistry, in general.The expression of CK19 was located in cytoplasm and cellular membrane.Gal-3 and HBME-1 were mainly detected in follicular luminal border and/or surface of papilla. The staining intensity in rapid immunohistochemistry was stronger than that in conventional immunohistochemistry. The positive rates of CK19,HBME-1,and Gal-3 by rapid immunohistochemistry in frozen sections were: 0 (0/28),10.7 % (3/28),0 (0/28),respectively,for benign lesions (nodular goiter,Hashimoto thyroiditis,thyroid adenoma); and 94.9 %(37/39),92.3 % (36/39),92.3 % (36/39),respectively,for PTC.The expression of three markers between thyroid benign lesions and PTC had a significant difference (x2 =59.326,55.861,44.605,all P < 0.001).In benign lesions,the rate of same case with two and more positive markers was 0,while in PTC it was 100 % and significantly different (x2 =67.000,P < 0.05).ConclusionMaxVision one-step rapid immunohistochemical staining technique could be applied in intraoperative frozen section diagnosis.Detecting CK19,HBME-1,and Gal-3 expression in intraoperative frozen section has an auxiliary value for diagnosis of PTC.
5.Comparison of OCTA and ICGA used in polypoid choroidal vasculopathy
Hou-Yin, XU ; Sheng-Kun, LANG ; Guo-Ge, HAN ; Bo, XIAO
International Eye Science 2017;17(7):1362-1364
AIM: To observe of optical coherence tomography angiography (OCTA) and indocyanine green angiography (ICGA) image feature in polypoidal choroidal vasculopathy (PCV).METHODS: Selected 21 patients 21 eyes with PCV in our hospital from January 2016 to December 2016.All the eyes were examined by ICGA, and was examined by OCTA after ICGA examination 1h.We observed the characteristics of OCTA and ICGA images.RESULTS:ICGA examination showed that there were 8 cases of choroidal abnormal branch vascular network (BVN), polypoid lesions 10 eyes, BVN with polypoid lesions 2 eyes, no abnormal performance 1 eyes.OCTA examination showed 8 eyes of BVN, and the location, range and shape of BVN were similar to ICGA in OCTA examination.ICGA examination showed 10 cases of polypoid lesions.OCTA showed strong signal highlights.ICGA examination showed 2 cases of BVN complicated with polypoid lesions, and OCTA examination showed strong signal highlights of BVN and corresponding parts.ICGA examination showed no abnormal performance in 1 eyes, and no abnormal findings in OCTA examination.CONCLUSION: OCTA and ICGA are similar in the location and morphology of PCV lesions, and OCTA may play a role in the diagnosis of PCV restricted ICGA.
6.Isolated peripheral neuropathy as an unusual presentation for an extramedullary relapse of acute leukemia
Xiao-Ying Zhu ; Sheng-Han Kuo ; Li-Ping Wan ; Ye Liu ; Yun-Cheng Wu
Neurology Asia 2014;19(2):203-206
A 23-year-old man in remission from acute myeloblastic leukemia after allogeneic peripheral blood stem cell transplantation developed peripheral neuropathy presenting as sciatic and peroneal nerve deficits. Electrophysiological tests localized the lesions to the left sciatic and common peroneal nerve. Magnetic resonance imaging revealed nerve thickening and enhancement, while a positron emission tomography-computed tomography scan demonstrated increased fluorodeoxyglucose uptake tracking along the nerve, suggesting peripheral nerve infiltration. This report demonstrates an unusual presentation of acute leukemia relapse presenting as focal neuropathy
8.Three-dimensional Hindlimb Gait Data Toolkit for Healthy and Spinal Cord Injured Rhesus Monkey
Wen ZHAO ; Jia-sheng RAO ; Can ZHAO ; Rui-han WEI ; Zhao-yang YANG ; Xiao-guang LI
Chinese Journal of Rehabilitation Theory and Practice 2015;21(7):760-765
Objective To establish a three- dimensional hindlimb gait data toolkit (THGT) for healthy and spinal cord injured (SCI) non-human primate (rhesus monkey) based on Matlab to realize upload of original data, automatic gait division, calculation and drawing of multiple gait parameters, etc. Methods Vicon system was used to collect three-dimensional hindlimb gait data of healthy and SCI (after 6 weeks) rhesus monkey to obtain the kinematics data of both hindlimbs in continuous strides. It was analyzed with THGT to process the gait division, calculation and drawing of multiple gait parameters. Results THGT read the data, distinguished cycles of gait, calculated 140 kinds of gait parameters and drew graphs of the results. Conclusion THGT extends the universality of the Vicon data, realizes automatically gait division and friendly interactive interface, and puts out the visible results.
10.Expressions of MMP-9 and TIMP-1 after focal cerebral ischemia-reperfusion in rats
He-Cheng CHEN ; Jin-Sheng YANG ; Xiao-Han DING ; Xiang-Qun SHI ; Yin-Sheng YANG
Chinese Journal of Neuromedicine 2008;7(7):661-664
Objective To explore the expressions of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of matrix metalloproteinase-1 (TIMP-1) after focal cerebral ischemia-repeffusion injury in rats. Methods Totally 40 Wistar rats were randomly divided into 2 groups: sham-operated group and iscbemia-reperfusion group. A middle cerebral artery occlusion model was constructed in the 25 rats with Longa's method. The expressions of MMP-9 and TIMP-1 were investigated with immunohistochemistry and HE staining at 6, 24, 48, 72 h and 7 d of reperfusion following 2 h ischemia. Results MMP-9 began to be expressed at 6 h reperfusion, and was obviously increased at 24 h and reached the peak level at 48 h, and then, the expression of MMP-9 began to decrease at 72 h to a low level at 7 d. TIMP-1 positive cells began to arise at 6 h reperfusion, peaked at 24 h, decreased at 48 h and remained a low level at 7 d. The expressions of MMP-9 and TIMP-1 were mainly located in vascular endothelial cells, neurons and gitter cells. The expressions of MMP-9 and TIMP-1 were negative in sham-operated group. Conclusion The expressions of MMP-9 and TIMP-1 are induced to increase by focal cerebral ischemia-reperfusion, and vascular endothelial injury may be the main cause to the high expressions of MMP-9 and TLMP-1.