1.Study on the cytokine network in the prevention of experimental autoimmune uveoretinitis by oral tolerance
Rui ZHANG ; Jiang QIAN ; Yifei YUAN
Chinese Journal of Ocular Fundus Diseases 2008;24(5):345-348
Objective To observe the interferon-γ(IFN-γ),interhukin-2(IL-2)levels of Th1 cytokine and IL-4、IL-10 levels of Th2 cytokine in serum and culture supernatants of splenic cells of the rats in the prevention of experimentaI autoimmune uveoretinitis(EAU)by oral tolerance. Methods 72 Lewis rats were randomly divided into EAU group,oral tolerance group(which including 10 μg、100 μg、1 mg、10 mg of S antigen group respectively)and control group,12 rats in each group.The animal model of EAU was induced by immunization with S antigen(50 μg)and Freund's complete adjuvant.Oral tolerance 10 μg、100μg、1 mg and 10 mg group were fed with 1 ml mixture of 10 μg、100μg、l mg、10 mg S antigen and 1 mg trypsin inhibitor respectively by intuhation,once the other day,totally 7 times,and then induced EAU according to above methods;control group was fed with 1 ml mixture of phosphate buffered saline and 1 mg trypsin inhibitor,once the other day,totally 7 times,and then induced EAU.The clinical manifestation of EAU in the eye were recorded,the eyeballs were enucleated at the peak of EAU,followed by pathological grading.Meanwhile the serum was colleced;splentic cells were separated and cultured to collect the supernatant.Cytokine levels of IFN-γ,IL-2,IL-4 and IL-10 in serum.cultured supernatant of splenic cells were determined by enzyme-linked immunosorbent assay(ELISA). Results Compared with EAU and control group,the levels of IFN-γ and IL-2(Th1 eytokine)in the serum in 100 μg and 1 mg group were decreased while the levels of IL-4 and IL-10(Th2 eytokine)were inereased,the diffefences were statistically significant(F=51.9,68.8,35.7,7.5,P<0.01).Compared the levels of Th1 and Th2 cytokines in the serum in 10 μg,10 mg group with EAU and control group.the differences were not statistically significant.In 100 gg、1 mg group,the levels of IFN-γ and IL-2(Th1 cytokine)in the culture supernatant of splenic cells were decreased while the Ievels of IL-4 and IL-10(Th2 cytokine)were increased,compared with EAU and control group,the differences were statistically significant(F=57.1,15.6,33.1,167.7,P<0.01).Compared the levels of Th1 and Th2 eytokine in the culture supernatant of splenic cells in 10 μg、10 mg groups with EAU and control group,the difference are not statistically significant. Conclusions In the process to prevent EAU by oral intake,the levels of IFN-γ and IL-2 (Th1 cytokine)were decrease while the levels of IL-4 and IL-10(Th2 cytokine).Oral administration with tOO high or low dose of the antigen can not prevent EAU as well as the cytokine levels do not change obviously.Cytokines has played an important role in the prevention of EAU.
2.Treatment of experimental autoimmune uveoretinitis with anti-tumor necrosis factor-α monoclonal antibody
Rui ZHANG ; Jiang QIAN ; Yifei YUAN
Chinese Journal of Ocular Fundus Diseases 2008;24(5):336-339
Objective To observe the efficacy of the anti-tumor necrosis factor-α monoclonal antibody(TNF-α MCAb)in the treatment of experimental autoimmune uveoretinitis(EAU). Methods EAU animal models were induced by interphotoreceptor retinoid-binding protein(IRBP)R16 peptide with immunization.The rats were divided into 2 groups according to the injection times.TNF-α MCAh was administered intravenously on day 6 or 4,6 and 8 post-immunization respectively,and then to observe the clinical expression by slit-lamp microscope.Meanwhile,take the rats which did not accept TNF-α MCAb as conttol group.Delayed type hypersensitivity(DTH)responses were measured on day 13 postimmunization of IRBP R16;the rats were killed on day 14 post-immunization of IRBP R16,and then enucleated the eyes for histopathological examination.To detect the cytokine level of IFN-γ,IL-4 in serum and IFN-γ in aqueous humor by enzyme-liked immunosorbent assay(ELISA)on day 14 postinjection.The hyperplasia responses of antigen specific lymphocyte of draining lymph node cells were detected. Results The TNF-αMCAb group had mitigated ocular inflammation and decreased pathological grades compared with the control group;the IFN-γ concentrations in aqueous humor and serum were decreased,IL-4 was increased in serum;DTH responses were decreased;the hyperplasia responses of draining lymphocytes to IRBP R16 peptide were decreased,all the differences were statistically significant(P<0.01).The rats accepted TNF-α MCAb thrice had much better curative effect than the rats injected once(P<0.05). Conclusions Injection of TNF-α MCAb can inhibit ocular inflammation and specific immune cells of EAU remarkably and change the Th1/Th2 balance.Many times injections of TNF-α MCAb were more effective than once.
3.Effects of Aurora A silence by RNA interference on the apoptosis and proliferation of glioma cells
Zhou XU ; Xianhou YUAN ; Pucha JIANG ; Kai FU ; Rui GONG
Acta Anatomica Sinica 2009;40(6):938-942
Objective To investigate the inhibitory effect of RNA interference on the expression of Aurora A in U251 cells, and the influence on proliferation and apoptosis of U251 cells. Methods The siRNA specific for Aurora A was synthesized and transfected into U251 cells in vitro. Aurora A mRNA expression and protein content were detected by RT-PCR and Western blotting respectively. The cell proliferation and apoptosis were observed by methyl thiazolyl tetrazolium(MTT) and flow cytometry(FCM). Transmission electron microscope was used to observe the ultrastructural changes of U251 cells. Results After transfection, the expression level of Aurora A mRNA was significantly decreased(P<0.01), and the protein content of Aurora A was also obviously reduced. The inhibitory rate of cell proliferation reached up to 67.57% 72 hours after transfection, which was significantly higer than that of normal control group(P<0.01). The apoptosis rate of U251 cells was significantly increased from (3.69±0.87)% to (15.34±2.16)% (P<0.01). Under the transmission electron microscope, it was observed that the U251 cells showed typical morphologic changes of apoptosis after transfection, such as karyopyknosis, chromatin condensation and margination, intracytoplasmic vacuoles formed, and apoptotic bodies formed. Conclusion The expression of Aurora A gene can be inhibited by siRNA successfully, and it results in the suppression of cell growth and induce apoptosis of human glioma cells in vitro. Aurora A may become a new target for the gene therapy of gliomas.
4.Relative of Th cytokines in MRL/Ipr mice——a model for idiopathic dacryoadenitis
Jie GUO ; Jiang QIAN ; Rui ZHANG ; Yifei YUAN
Chinese Journal of Microbiology and Immunology 2009;29(9):782-785
Objective To determine the distribution of IL-4, IL-17, IFN-γ in the lacrimal gland lesions and submaxillary lymph nodes of MRL/Ipr mice-a model for idiopathic dacryoadenitis, and investi-gate the possible role of T-belper cells in the disease. Methods Frozen sections of lacrimal glands and sub-maxillary lymph nodes from MRL/lpr and BALB/c mice age 3 months were stained with antibodies to the cy-tokines IL-4, IL-17 and IFN-γ, then calculated the numbers or proportions of positive cells respectively,double staining for CD4 and each cytokine was taken in lymph nodes of MRL/Ipr mice additionally. Results The major cytokine in lacrimal gland lesions is IL-4, which contrasts to little IL-17 or IFN-γ/stained(P <0.05), the numbers of cells staining for each cytokine in submaxillary lymph nodes elevate in MRL/lpr mice, and the expressions of IL-4 and IL-17 are greater than IFN-γ significantly. Conclusion Th2 may play an important role in the development of local dacryoadenitis and systemic abnormality in MRL/lpr mice,the role of IL-17 maybe different in lacrimal gland lesions and lymph nodes which need more research.
5.Evaluation of spinal cord vessels using multi-slice CT angiography
Shuang CHEN ; Rui-Jiang ZHU ; Xiao-Yuan FENG ;
Chinese Journal of Radiology 2001;0(08):-
Objective To evaluate the value of Multi-slice spiral CT angiography for spinal cord vessels.Methods 11 adult subjects with suspected of myelopathy were performed with Multi-slice spiral CT angiography,An iodine contrast agent was injected at 3.5 ml/s,for total 100 ml.The parameters were axial 16 slice mode,0.625 mm slice thickness,0.8 s rotation,delay time depending on smartprep (15—25 s), multi-phase scan.The coronal and sagittal MPR and SSD were generated on a workstation compared with spinal digital subtraction angiography (DSA) to analyze normal or abnormal spinal cord vessels.Results Normal findings at spinal CTA and digital subtraction angiography in six adult normal subjects and spinal cord vascular malformations( 1 intradural extramedullary AVF,4 dural AVFs) in five cases,Recognizable intradural vessels corresponding to anterior median (midline) veins and/or anterior spinal arteries were show in six adult normal subjects.Abnormal intradural vessels were detected in all five spinal cord vascular malformation with CT angiography ,in comparison with digital subtraction angiography these vessels were primarily enlarged veins of the coronal venous plexus on the cord surface,radiculomedullary-dural arteries could not be clearly shown in four dural AVF,only one anterior spinal artery was detected in one patient with intradural medullary AVF,which direct shunt between anterior spinal artery and perimedullary vein with tortuous draining vessel.Conclusion Multi-slice CT angiography is able to visualize the normal or abnormal spinal cord vessels.It could be used as a noninvasive method to screen the spinal cord vascular disease.
6.Influence of long-time illumination of monochromatic light on density of cones and opsin expression in guinea pig
Min, HU ; Yong-song, LIU ; Rui, LIU ; Chun-guang, JIANG ; Ren-yuan, CHU ; Yuan-sheng, YUAN
Chinese Journal of Experimental Ophthalmology 2011;29(3):196-201
Background The visual system of animal have to optimally adjust in various environmental conditions in order to obtain stable and effective visual funetion.However,the color vision system of animals which encounter uncertainty of spectral signals should be plastic.Whether the densities of various cones and expression of opsins change with long-time spectral deprivation is unclear.Objective This study was to investigate the changes of cone density as well as the expression of corresponding opsin and mRNA following the long-term illumination of monochromatic light.Methods Thirty 3-day-old guinea pigs were randomized into 3 groups and exposed tO the 530 nm green light,400 nm purple light and white light for consecutive 8 weeks respectively.The flat-mounted retinal sample was prepared and divided into dorsal zone,ventral zone and mixed zone anatomically according to the distribution of difierent light-sensitive cone.The changes in density of cone cells sensitivited to different colored light were detected by single-1abel or double-label immunocytochemistry.The levels of opsin and its mRNA were determined using Western-blot and real-time PCR respectively.Results The density of green-sensitivity cones was significantly different in the dorsal zone of retina among green light group,purple light group and white light group (F=234.28,P<0.01).Compared with white light group,the density of green-sensitive cones in dorsal retina of green light group was obviously higher but that of purple light group wag evidently lower(q=389.68,P<0.01;q=67.11,P<0.01).No significant difference was found in the density of purple-sensitive cones in the ventral zone of retina among green light group,purple light group and white light group(F=3.14,P>0.05).The density of coexpression of the mixed cone cells was increased in green light group(q=157.55,P<0.01)but decreased in purple light roup (q=254.85,P<0.01)in comparison with white light group.The expression levels of green-opsin and green-opsin mRNA in green light group was significantly elevated(q=184.45,P<0.01;q=4.71,P<0.05),but those of purple light group were evidently declined(q=5.87,P<0.05;q=346.66,P<0.01)in comparison with white light group.There was no statistically significant differences were found in the expression of purple-opsin and its mRNA among all the groups(F=1.24,P>0.05;F=3.27,P>0.05).Conclusion After the exposure of long-time monochromatic light illumination,monochromatic cones density and its opsin in guinea pig occur the corresponding alteration to gain good spatial vision as a compensatory reaction.These outcomes imply that there is some plasticity during the development of color vision.The increase of green-sensitive cones might be from the differentiation of coexpression cones in transition region.
7.Development of a Rapid System for Bacteria Identification in 24 Hours
Xianbin LI ; Rui ZHAO ; Caizhi HUANG ; Yuan LAI ; Yulian JIANG ; Sai LI ; Xunliang JIANG
Chinese Journal of Nosocomiology 2006;0(09):-
OBJECTIVE To develop a rapid system for bacteria identification and susceptibility test in 24 hours,and provide bacteriological evidence for the control of nosocomial infection and the timely diagnosis and treatment.METHODS The simple constant temperature box was replaced by revolving constant temperature box;2,3,5-triphenyltetrazolium chloride(TTC) and succinate were added to the culture bottle and the culture medium of drug susceptibility;the concentration of the reactive substrate in the bacterial biochemical tube and the number of the inoculated bacteria were increased at the same time.RESULTS The time of positive blood culture in the revolving constant temperature box was significantly shorter than that in the simple one(?2=74.92,P
8.Clinical study of tamsulosin in prevention of urinary retention after radical hysterectomy
Xueling QU ; Jiyong JIANG ; Rui CAO ; Lili LIU ; Kun GAO ; Yuan LUO
Chinese Journal of Postgraduates of Medicine 2010;33(9):18-19
Objective To study the clinical value of tamsulosin in prevention of urinary retention after radical hysterectomy.Methods Sixty-six patients after radical hysterectomy were enrolled in this study, and divided into two groups in random.Thirty-three cases in the treatment group were treated with tamsulosin 0.2 mg once every night,3 days before the catheter was drawn.While 33 cases in the control group only with placebo.Catheter keeping days, residual urine volume at first extraction,the rate of urinary retention and urinary tract infection,hospitalization time and drug adverse reaction were compared in two groups.Results Catheter keeping days and hospitalization time were shorter in the treatment group than those in the control group(P< 0.05).The residual urine volume at first extraction was(38.2 ± 5.6) ml in the treatment group and (168.5 ± 11.8) ml in the control group,there was significant difference between two groups (P< 0.05).The rate of urinary retention was 18.2%(6/33) in the treatment group and 36.4%(12/33) in the control group,there was significant difference between two groups (P <0.05).There was no adverse reaction resulted from tamsulosin in the treatment group.Conclusion Tamsulosin is an effective drug which could prevent and treat the urinary retention after radical hysterectomy.
9.The diversity of endophytic fungi in endangered medicinal material Daphnes Cortex as determined by high-throughput sequencing technology
Yuan HE ; Qi-rui MU ; Zhen-fang BAI ; Dan JIANG ; Guang-xi REN ; Chun-sheng LIU
Acta Pharmaceutica Sinica 2021;56(3):872-878
In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%.
10.Effects of 5-Aza-CdR on the proliferation of human breast cancer cell line MCF-7 and on the expression of Apaf-1 gene.
Huihua, XIONG ; Hong, QIU ; Liang, ZHUANG ; Hua, XIONG ; Rui, JIANG ; Yuan, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):498-502
Hypermethylation in the promoter region of tumor suppressor genes is a common mechanism of gene silencing, which tends to occur in cancer. The effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR), a specific DNA methyltransferase inhibitor, on the cell proliferation of human breast cancer cell line MCF-7 and on the expression of Apaf-1 gene were investigated. Human MCF-7 cells were incubated with increasing concentrations of 5-Aza-CdR for 12 to 120 h. The growth inhibition rates of MCF-7 cells were detected by MTT assay. Changes of cell cycle distribution and apoptotic rates of MCF-7 cells were determined by flow cytometry. The expressions of DNA methyltransferase 3b mRNA and Apaf-1 mRNA were measured by reverse transcription polymerase chain reaction (RT-PCR). Meanwhile, the expression of Apaf-1 protein was detected by Western blotting. The results showed that 5-Aza-CdR significantly inhibited the growth of MCF-7 cells and the growth inhibition rate of MCF-7 cells was significantly enhanced with the concentration of 5-Aza-CdR and the action time. Flow cytometry indicated that 5-Aza-CdR could significantly induce G(1)/S cell cycle arrest and increase the apoptosis rate of MCF-7 cells. The mRNA and protein expressions of Apaf-1 were up-regulated in MCF-7 cells treated with 5-Aza-CdR, which was accompanied by down-regulation of DNA methyltransferase 3b mRNA. It is concluded that 5-Aza-CdR might retard the growth of tumor cells and promote the apoptosis of MCF-7 breast cancer cells by inhibiting the expression of DNA methyltransferase 3b and re-activating the Apaf-1 gene expression.