1.Expression of interleukin-17 and interleukin-22 in supernatant of peripheral blood mononuclear cell and tear in dry eye patient
Xu-hua, TAN ; Song, SUN ; Ting-ting, ZHU ; Tian-li, REN ; Zhi-feng, WU ; Ke, WANG
Chinese Journal of Experimental Ophthalmology 2013;(7):668-673
Background T helper cell 17 (Th17),a newly discovered subset of CD4+ T cells,have been found to play an important role in dry eye disease in animal model.Further investigation should be done on the immunopathogenesis of Th17 cells in dry eye patients.Objective This study was to analyze the expression status of interleukin-17 (IL-17) and IL-22 in tear and supernatant of peripheral blood mononuclear cells (PBMCs) of dry eye patients and their correlation with clinical symptom and sign.Methods Twenty Sj(o)gren syndrome (SS)patients,twenty non-Sj(o)gren syndrome (NSS) patients were included in Wuxi Second Hospital from 2010 to 2011,and twenty healthy volunteers were recruited at the same period.All of subjects understood the purpose and procedure of research and written informed consent was obtained form each subject initial of this study.Dry eye symptom questionnairs were self-answered and multiple dry eye disease-related clinical tests,including the breakup time of tear film (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescein staining were performed.The periphery blood of 3 ml and tear were collected in all the subjects,and IL-17 and IL-22 levels in supernatant of PBMCs and tear were detected by enzyme-linked immunosorbent assay (ELISA).The correlations between levels of IL-17 and IL-22 with BUT,S Ⅰ t,corneal fluorescein staining and dry eye scores were analyzed.Results The dry eye scores reduced,BUT prolonged,S Ⅰ t increased and corneal fluorescein dye decreased from SS group,NSS group to normal control group,with significant differences among the three groups (dry eye scores:H =40.81,P<0.01 ; BUT:H =40.15,P<0.01 ;S Ⅰ t..H=50.07,P<0.01 ;corneal dye scores:H=40.52,P<0.01).The concentration of IL-17 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (964.92±124.83)ng/L,(718.85± 115.89)ng/L and (341.95±85.08) ng/L,showing a statistically significant difference among them (F=162.95,P<0.01).The levels of IL-17 in the tear were (440.69±126.09) ng/L,(364.33±126.85) ng/L and (61.16±11.60) ng/L in the SS group,NSS group and normal control group respectively,exhibiting an elevated level in the SS group and NSS group compared with the control group (F=75.27,P<0.01).In addition,the levels of IL-22 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (98.77± 11.27) ng/L,(79.65 ± 11.01) ng/L and (32.78±9.34) ng/L,and those in the tear were (22.22 ± 8.96) ng/L,(14.92 ±4.35) ng/L and (10.47 ± 2.67) ng/L,with significant differences among the three groups (F =206.27,P<0.01 ;F =19.87,P<0.01).The significant correlations were found between the IL-17 and IL-22 concentration in the supernatant of PBMCs and tear with corneal fluorescein staining scores and S Ⅰ t.Conclusions The contents of IL-17 and IL-22 in PBMCs and tear upregulate in the SS and NSS patients,indicating that Th17 plays a key role in the immunity mechanism of dry eye.
2.Clinical analysis of interleukin-6 level in tear and serum of dry eye patient
Song, SUN ; Xu-hua, TAN ; Tian-li, REN ; Zhi-feng, WU ; Ting-ting, ZHU ; Hui-yan, XU
Chinese Journal of Experimental Ophthalmology 2013;(2):186-190
Background Mechanism of dry eye disease has not been established clearly.Some increasing evidences showed that inflammation plays an important role in pathogenesis and development of dry eye disease.However,the association of interleukin-6 (IL-6) with dry eye has not been clarified.Objective This study was to assess the correlation of IL-6 levels in tear and serum of patient with dry eye disease and clinical symptom.Methods Twenty patients with Sj(o)gren syndrome (SS) and 20 cases with non-Sj(o)gren syndrome (NSS) were enrolled from Second Affiliated Hospital of Nanjing Medical University,and 20 healthy volunteers were recruited as the normal controls.Symptom questionnaires were self-answered and multiple dry eye-related clinical tests were performed,including tear film breakup time (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescence staining.IL-6 concentrations in serum and tear were detected by enzyme-linked immunosorbent assay (ELASA).The correlation between IL-6 level and tear film and ocular surface parameters was analyzed by Spearman rank correlation.Written informed consent was obtained from each subject initial of this trial.Results The scores of symptom questionnaires were 18.50 (11.75,23.00),23.00 (19.00,32.00) and 2.00 (0.25,4.00) in the SS group,NSS group and the normal control group,and significantly increasing values were seen in the SS group and NSS group compared with the normal control group (P=0.00,0.00).BUT values in the SS group and NSS group were 2.50 (2.00,3.88) seconds and 3.50 (2.13,4.00) seconds and were lower than 15.00 (13.13,17.00) seconds in the normal control group (P=0.00,0.00).S Ⅰ t values in the SS group,NSS group and the normal control group were 2.00 (1.50,2.88),4.00 (3.00,5.75) and 19.00 (18.00,25.00)mm,showing significant differences between the SS group and the normal control group (P =0.00),the SS group and the NSS group (P =0.00) or the NSS group and the normal control group (P=0.00).The marked elevation of keratoepithelioplasty score was seen between the SS group and the normal control group (5.75 (4.00,7.75) vs.0.00 (0.00,0.75) (P =0.00) or the NSS group and the normal control group (4.50(3.63,6.00) vs.0.00(0.00,0.75)) (P=0.00).Concentrations of IL-6 in serum and tear in the SS group,NSS group were (131.47±21.04) μg/L and (77.81 ± 15.68) μg/L and were significantly higher than (31.62±8.57) μg/L of the normal control group,and those in the tear were (33.44±8.01) μg/L,(18.78 ±5.73) μg/L and (8.77 ± 3.43)μg/L,respectively in the three groups,with a significant reduce in the normal control group (all P=0.00).tL-6 levels in serum and tear were positively correlated with the score of symptom questionnaires or keratoepithelioplasty and negatively correlated with BUT and S Ⅰ t (all P =0.00).Conclusions IL-6 levels in the tear and serum of the dry eye patient are associated with the severity of the disease.IL-6 levels in serum and tear may be objective,sensitive and reliable indicators of dry eye.
3.Characterization of surface EMG signals using improved approximate entropy.
Wei-ting CHEN ; Zhi-zhong WANG ; Xiao-mei REN
Journal of Zhejiang University. Science. B 2006;7(10):844-848
An improved approximate entropy (ApEn) is presented and applied to characterize surface electromyography (sEMG) signals. In most previous experiments using nonlinear dynamic analysis, this certain processing was often confronted with the problem of insufficient data points and noisy circumstances, which led to unsatisfactory results. Compared with fractal dimension as well as the standard ApEn, the improved ApEn can extract information underlying sEMG signals more efficiently and accurately. The method introduced here can also be applied to other medium-sized and noisy physiological signals.
Algorithms
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Cluster Analysis
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Data Interpretation, Statistical
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Electromyography
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methods
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Entropy
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Fractals
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Humans
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Models, Statistical
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Nonlinear Dynamics
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Pattern Recognition, Automated
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Signal Processing, Computer-Assisted
4.Expression of a testis-specific gene 1700001022RIK in mice and its bioinformatic analysis.
Yu-chi LI ; Shou-ren LIN ; Man-ling LUO ; Huan GUO ; Han-wei WU ; Zhi-mao JIANG ; Yao-ting GUI
National Journal of Andrology 2015;21(5):391-395
OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.
METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.
RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.
CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.
Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis
5.Buyang Huanwu decoction promotes neuroblast migration from subventricular zone via inducing angiogenesis after ischemia.
Lin LI ; Zhi-ting LIU ; Li-sheng CHU ; Tian-hong YU ; Tie-bing QU ; Jun WANG ; Cui-cui REN
China Journal of Chinese Materia Medica 2015;40(2):298-302
OBJECTIVETo study the effect of Buyang Huanwu decoction (BYHWD) inducing angiogenesis on the neuroblast migration from the subventricular zone and its mechanisms after focal cerebral ischemia.
METHODThe middle cerebral artery occlusion (MCAO) was performed to mice for 30 minutes to establish the model. The rats were divided into sham group, model group, BYHWD group and endostatin group. BYHWD (20 g x kg(-1), ig) and endostatin (10 μg, sc) were administered 24 h after ischemia once a day for consecutively 14 days. At 14 d after ischemia, the density of micro-vessel and the number of neuroblasts in the ischemia border zone were determined by immunofluorescence staining. The mRNA and protein expression of cell-derived factor-1 (SDF-1) and brain-derived neurotrophic (BDNF) were examined by real-time PCR and Western blot.
RESULTCompared with the model group, BYHWD significantly increased the density of micro-vessel and the number of DCX positive cells in the ischemia border zone (P < 0.01), and significantly increased the SDF-1 and BDNF mRNA and protein expression (P < 0.01). Compared with BYHWD group, endostatin significantly reduced the density of micro-vessel and the number of DCX positive cells in the ischemia border zone (P < 0.01), as well as the SDF-1, BDNF mRNA and protein expression (P < 0.01).
CONCLUSIONBYHWD could promote the neuroblast migration from the subventricular zone via inducing angiogenesis after cerebral ischemia, the mechanism may be correlated with up-regulating the expression of SDF-1 and BDNF.
Angiogenesis Inducing Agents ; pharmacology ; Animals ; Brain Ischemia ; pathology ; physiopathology ; Brain-Derived Neurotrophic Factor ; analysis ; genetics ; Cell Movement ; drug effects ; Cerebral Ventricles ; pathology ; Chemokine CXCL12 ; analysis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Neurons ; drug effects ; physiology
6.Analysis of the GSTP1 gene Ile105Val polymorphism in fluorosis areas in Guizhou Province
Ting, ZHANG ; Ke-ren, SHAN ; Dong, AN ; Shi-qing, XU ; Shu-guang, ZHOU ; Yan, HE ; Chang-xue, WU ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2009;28(3):268-271
Objective To investigate plasma glutathione S-transferase(GSTs) activity and GSTP1 gene Ile105Val polymorphism in Bijie City, Guizhou Province, a coal-burning fluorosis endemic area. Methods One hundred and sixty villagers from Yachi Twon using non-improved cooking stoves were selected as the non-intervened group in Bijie City, Guizhou Province where coal-burning fluorosis was prevailing; 153 villagers as the intervented group were chosen from Changchun Twon, where cooking stoves were improved; 151 villagers were served as the control group from Baiyunshan Twon, Changshun County without endemic fluorosis. The activity of GSTs was tested by colorimetric analysis with spectrophotometer. The genotype of the GSTP1 gene Ile105Val polymorphism, presenting as either homozygous wild-type (AA), or heterozygous mutation type (AG), or homozygous mutation type (GG), was detected through the PCR-RFLP procedure. Results The activity of GSTs in plasma of non-intervened group [(12.44±4.97) kU/L]was significantly lower than that of intervened group (P < 0.05), and that of intervened group[(20.78±6.20)kU/L]was significantly lower than that of control group[(24.30±6.27)kU/L, P< 0.05]. The difference of the enzyme activity of three groups were statistically significant (F = 51.71, P < 0.05), but this enzyme activity did not vary significantly in each sex of each grnup(P > 0.05). Compared intervened group [AA:67.3%(103/153), AG:29.4%(45/153),GG:3.3%(5/153)]and non-intervened group[AA:66.9%(107/160), AG:30%(48/160), GG:3.1%(5/160)]with control group[AA:74.8%(113/151), AG:25.2%(38/151), GG:0 (0/151)], the Ile105Val polymorphism site of GSTP1 gene had significant difference(χ2= 6.04,6.07, both P< 0.05), but not significant between intervened and non-intervened groups(χ2 = 0.02, P>0.05). Conclusions Fluorosis can decrease the activity of GSTs and introduce the GSTP1 gene Ile105Val polymorphism, intervention with the fluorine intake will improve the effect of fluoride on the body.
7.Augmentative plate fixation for the treatment of femoral hypertrophic nonunions subsequent to intramedullary nailing fixation.
Jian-Zheng ZHANG ; Zhi LIU ; Tian-Sheng SUN ; Jing-Sheng LI ; Ji-Xin REN ; Shu-Qing LIU ; Shao-Ting XU
China Journal of Orthopaedics and Traumatology 2010;23(12):932-935
OBJECTIVETo investigate the effect of augmentative plate fixation to increase stability in the treatment of femoral shaft nonunions subsequent to intramedullary fixation.
METHODSNine patients with femoral nonunions after intramedullary nail internal fixation were treated with augmentative plate internal fixation from April 1998 to Jane 2008, included 8 males and 1 female, with an average age of 32 years old ranging from 21 to 54 years. One case was upper 1/3 femoral fractures, 5 cases were middle 1/3 femoral fractures, 3 cases were lower 1/3 femoral fractures. The interspace of bone nonunion was more than 5 mm in 6 cases, of them, iliac bone grafting were applied in 4 cases, artificial bone combined with iliac bone grafting were applied in 2 cases; The interspace of bone nonunion was less than 5 mm in other 3 cases,artificial bone grafting was applied in 1 case, fitting bone callus were applied in 2 cases. All patients got protected weight loading preventing the main screw break.
RESULTSAll patients achieved radiological solid union at an average of 8 months (ranged 6 to 11 months ). The fixation was removed during 6 to 11 months after operation in 5 cases. Donor site pain of iliac occurrenced on 4 cases,3 cases relieved 1 month later and 1 case relieved 3 months later. No infection, fixation loosening or breaking was observed.
CONCLUSIONThe augmentative plate fixation can be applied at the fracture site to prevent the rotational instability. The technique is simple and does not require any special instrument, which facilitates an early weight bearing and gives a quick recovery from nonunion.
Adult ; Bone Plates ; Female ; Femoral Fractures ; surgery ; Femur ; pathology ; Fracture Fixation, Intramedullary ; methods ; Fractures, Ununited ; surgery ; Humans ; Hypertrophy ; Male ; Middle Aged
8.A study of interleukin-10 gene polymorphisms in Miao, Dong and Buyi ethnics of Guizhou.
Chan-juan WANG ; Ke-ren SHAN ; Yan HE ; Ting ZHANG ; Yi LI ; Chang-xue WU ; Chan ZHANG ; Zhi-zhong GUAN
Chinese Journal of Medical Genetics 2013;30(1):116-120
OBJECTIVETo investigate allelic frequencies of interluekin-10 (IL-10) gene promoter in Miao, Dong and Buyi ethnics of Guizhou.
METHODSTaqMan MGB-based real-time PCR was used to determine the genotypes of IL-10 -819 and IL-10 -592 in 589 Miao, Dong and Buyi ethnics of Guizhou.
RESULTSThe allelic frequency of IL-10 -819 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. Allelic frequencies of IL-10 -592 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. In Miao, Dong and Buyi ethnics, the distributions of genotype frequencies of IL-10 -819 and IL-10 -592 were statistically different from Han ethnics from Guizhou and Taiwan of China as well as South Koreans.
CONCLUSIONThere is a heterogeneity in the frequencies of polymorphisms of IL-10 promoter among different ethnic groups.
Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Interleukin-10 ; genetics ; Polymorphism, Single Nucleotide ; Population Groups ; genetics ; Promoter Regions, Genetic
9.Expression characteristics of the Ccdc70 gene in the mouse testis during spermatogenesis.
Jian-bo CHEN ; Wen-zhong ZHENG ; Yu-chi LI ; Shou-ren LIN ; Zeng ZHANG ; Yong WU ; Zhi-mao JIANG ; Yao-ting GUI
National Journal of Andrology 2016;22(1):12-16
OBJECTIVETo investigate the expression characteristics of the gene of coiled-coil domain-containing protein 70 (Ccdc70) in the mouse testis and its potential role in spermatogenesis.
METHODSUsing expression profile microarray, we screened the mouse testis-specific gene Ccdc70, studied its expression characteristics in the mouse testis by RT-PCR, real-time PCR, Western blot and immunohistochemistry, followed by bioinformatic analysis of the Ccdc70 protein.
RESULTSThe Ccdc70 gene was expressed highly in the testis but lowly in the epididymis of the mice. The Ccdc70 protein was expressed mainly in the spermatocytes and round spermatids of the testis and in the epithelial cells of the epididymis. Bioinformatic analysis showed a structural domain in the Ccdc70 protein, which was highly conserved in mammalian evolution.
CONCLUSIONThe Ccdc70 gene is highly expressed in the mouse testis and mainly in the spermatocytes, round spermatids, and epididymal epithelial cells, which indicates that it is involved in the regulation of spermatogenesis and epididymal sperm maturation.
Animals ; Computational Biology ; Gene Expression Regulation, Developmental ; Male ; Mice ; Proteins ; genetics ; Spermatogenesis ; genetics ; Testis ; metabolism
10.Establishment of inflammatory model induced by Pseudorabies virus infection in mice
Chun-Zhi REN ; Wen-Yue HU ; Jin-Wu ZHANG ; Ying-Yi WEI ; Mei-Ling YU ; Ting-Jun HU
Journal of Veterinary Science 2021;22(2):e20-
Background:
Pseudorabies virus (PRV) infection leads to high mortality in swine. Despite extensive efforts, effective treatments against PRV infection are limited. Furthermore, the inflammatory response induced by PRV strain GXLB-2013 is unclear.
Objectives:
Our study aimed to investigate the inflammatory response induced by PRV strain GXLB-2013, establish an inflammation model to elucidate the pathogenesis of PRV infection further, and develop effective drugs against PRV infection.
Methods:
Kunming mice were infected intramuscularly with medium, LPS, and different doses of PRV-GXLB-2013. Viral spread and histopathological damage to brain, spleen, and lung were determined at 7 days post-infection (dpi). Immune organ indices, levels of reactive oxygen species (ROS), nitric oxide (NO), and inflammatory cytokines, as well as levels of activity of COX-2 and iNOS were determined at 4, 7, and 14 dpi.
Results:
At 105 –106 TCID50 PRV produced obviously neurological symptoms and 100% mortality in mice. Viral antigens were detectable in kidney, heart, lung, liver, spleen, and brain. In addition, inflammatory injuries were apparent in brain, spleen, and lung of PRVinfected mice. Moreover, PRV induced increases in immune organ indices, ROS and NO levels, activity of COX-2 and iNOS, and the content of key pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6, tumor necrosis factor-α, interferon-γ and MCP-1. Among the tested doses, 10 2 TCID 50 of PRV produced a significant inflammatory mediator increase.
Conclusions
An inflammatory model induced by PRV infection was established in mice, and 102 TCID50 PRV was considered as the best concentration for the establishment of the model.