ObjectiveTo determine the signal pathway of specifically expressed oncostatin M(OSM) in lens inducing retinal degeneration in transgenic mice.MethodsA sequence-truncated OSM cDNA (661 bp) of mice was linked to αA-crytallin promoter, and was micro-injected into unicellular embryo to set up the model of transgenic mice. Reversal transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of gp130/OSMRβ receptor in the retinae of OSM transgenic and non-transgenic mice. Rabbit anti-phosphorylated STAT-3 antibody was used to detect the protein expression of phosphorylated STAT-3，and mouse anti-cytochrome C antibody was used to detect the distributing of cytochrome C in retinae. ResultsExpression of gp130/OSMRβmRNA was found in retina of non-transgenic mice. At the 17.5th day in the embryonic stage, significant accumulation of the phosphorylated STAT-3 was detected in the retinal nucleolus in OSM transgenic retina. At the first day after birth, intensive staining of cytochrome C in OSM transgenic retina was found. Conclusionsspecifically expressed OSM in lens may act on gp130/OSMRβ receptor in retinae, activate STAT-3, and cause the release of cytochrome C from mitochondria, which eventually induces widespread retinal degeneration.(Chin J Ocul Fundus Dis, 2005,21:167-169)
【摘要】 目的 探讨肝炎后肝硬化自发性细菌性腹膜炎（spontaneous bacterial peritonitis，SBP）的诊疗情况及头孢哌酮舒巴坦联合左旋氧氟沙星对SBP的治疗效果。 方法 对2004年1月-2009年12月收治的54例肝炎后肝硬化SBP患者，应用头孢哌酮舒巴坦联合左旋氧氟沙星给与治疗，并观察分析治疗效果。 结果 肝炎后肝硬化SBP的临床表现以发热，腹痛为主，具有典型腹膜刺激征的不足半数。外周血白细胞升高者不多见，腹腔积液中白细胞计数、PMN计数和细菌培养是自发性细菌性腹膜炎的重要诊断指标。 结论 肝炎后肝硬化合并SBP的临床表现不典型。致病菌以G-杆菌为主。在早期诊断、综合治疗的基础上，头孢哌酮舒巴坦联合左旋氧氟沙星对自发性细菌性腹膜炎的治疗效果显著。【Abstract】 Objective To evaluate the diagnosis and treatment of post-hepatitis cirrhosis complicated with spontaneous bacterial peritonitis (SBP) cases and the efficacy of cefoperazone and sulbactam combined with levofloxacin in the treatment of cirrhotic patients with SBP. Methods From January 2004 to December 2009, the clinical data from 54 cases of SBP after cirrhosis were analyzed. The patients underwent the treatment of cefoperazone sulbactam combined with levofloxacin. The therapeutic effect was observed. Results The main clinical manifestations were fever and abdominal pain, and about half of the patients had the typical peritoneal irritation. Only a few patients had elevated peripheral white blood cells (WBC). The WBC count, abdominal effusion polymorphonuclearcyte count and bacteria cultivation were the indexes of diagnosis of SBP. Conclusion The clinical features of post-hepatitis cirrhosis complicated with SBP are not typical. The main pathogenic bacteria is G- bacilli. In the early diagnosis and treatment, cefoperazone sulbactam combined with levofloxacin is effective.
Objective To explore the feasibility of allogeneic marrow stromal stem cells(MSCs) as seed cells to construct tissue engineered bone bydetecting the expressions of interleukin 2(IL-2) and IL-2 receptor in rhesus monkeys after implanting these tissue engineered bones.Methods Engineered bones were constructed with osteoblasts which derived from allogeneic MSCs and bio-derived materials in vitro, and then were implanted to bridge 2.5 cm segmental bone defects of left radius in 15 rhesus monkeys as experimental group, bioderived materials only were implanted to bridge same size defects of right radius as control group. Every 3 monkeys were sacrificed in the 1st, the 2nd, the 3rd, the 6th andthe 12th weeks postoperatively and the expressions of IL-2 and IL-2 receptor in blood and graft samples were detected quantitatively by enzymelinked immuneosorbent assay (ELISA).Results There was no significant difference in the contents of IL-2 and its receptor between 2 groups（P＞0.05）. The contents ofIL-2 and its receptor increased from the 2nd week and maintained high level from the 2nd to the 6th week, but decreased after 6 weeks.ConclusionTissue engineered bones constructed with allogeneic MSCs and bio-derived materials show low immunogenicity. Allogeneic MSCs may be used as seed cells to construct tissue engineered bone.
ObjectiveTo investigate the role of interleukin-25 (IL-25) and its receptor during allergen challenge test in allergic asthmatics as well as its underlining mechanism.MethodsFifteen allergic asthmatic patients with dual response in allergen challenge test were enrolled and blood samples were collected before and after challenge test. The expression levels of IL-25 receptor on the surface of eosinophils, plasma and intracellular IL-25 levels were measured by flow cytometry and enzyme-linked immunosorbent assay. Besides, the function of eosinophils from these patients was evaluated through the expression of type 2 cytokines, degranulation and chemotaxis after stimulation with IL-25.ResultsUpon allergen challenge, the expression of IL-17RB on the surface of eosinophils were increased from (7 426±2 824)/106 white blood cells to (19 446±5 593)/106 white blood cells (P<0.001). The expression of IL-17RA/RB on eosinophils were significantly increased from (4 508±1 360)/106 white blood cells to (9 025±3 166)/106 white blood cells (P<0.001). The plasma level of IL-25 increased from (650±45) pg/ml to (851±43) pg/ml (7 hours after allergen challenge) and (813±56) pg/ml (24 hours after allergen challenge) (P<0.001). The intracellular IL-25 expression of eosinophils was also upregulated from (10 398±1 909)/106 white blood cells to (147 684±46 222)/106 white blood cells (P<0.05). In vitro study, IL-25 (1 ng/ml) stimulated eosinophils for 2 hours promoted its expression of peroxidase [(12.5±4.2) ng/ml compared to control (1.26±0.4) ng/ml, P<0.05). The intracellular expression of IL-5 and IL-13 in eosinophils were also increased after stimulated by IL-25. IL-25 (1 pg/ml) stimulation compared to control could increase eosinophil migration in eotaxin [(36±3) vs. (69±5), P<0.05).ConclusionIL-25 and its receptor play a critical role in eosinophilic aggregation, activation and mobilization during allergic inflammation in allergic asthmatics.
Purpose To determine the effect of exogenous interleukin-1alpha; (IL-1alpha;) on the retina and its vasculature and VEGF expression in SD rats． Methods IL-1alpha;2．0 ng (20 mu;l) were injected into the vitreous of 8 left eyes of 8 SD rats while steriled PBS were injected into 8 right contralateral eyes of the same rats as control． All eyes were assessed by direct ophthalmoscopy every day and enucleated on the 7 thpostoperative day． Histological examination (hemato xylineosin staining) and immunohistochemical staining with antibody against VEGF antigen were performed， and sections were observed and photographed under light microscopy． Results ①All 8 IL-1alpha; inject ed eyes developed epiretinal membranes and extraretinal neovascularization on the 3 rd postoperative days while none of the 8 control eyes exhibited any a bnormal retinal vascular changes and they were confirmed by HE staining;②Immuno staining identified VEGF express mainly in the inner layer of vessel walls， the epiretinal membranes， the neuroganglional layer and the photoreceptor layer of retina， while the control eyes showed only weak positive staining in the photo receptor layer． Conclusions IL-1alpha; is capable of inducing vitreo retinal neovascularization，and increasing the expression of VEGF in the retina and epiretinal membranes． (Chin J Ocul Fundus Dis, 2001,17:135-137)
Objective To construct a regulatable plasmid containing single chain fusion gene of murine interleukin-12 (mIL-12) which was regulated with mifepristone (RU486) and explore its expression in vitro. Methods The p40 and p35 subunit sequence of mIL-20 were respectively obtained from the plasmid GCp35Ep40PN by polymerase chain reaction (PCR) and they were cloned into pCA14 plasmid after introducing a linker by overlap PCR. The single chain mIL-12 gene was comfirmed by sequencing and subcloned into pRS-17 vector which contains RU486 regulator cassette. The positive clone named pRS-RUmIL-12 was identified by restriction endonuclease digestion and PCR. Lipofectamine 2000 was used to transfect the pRS-RUmIL-12 to HEK293 cells followed by manufacturer’s recommendations. The protein concentration of mIL-12 induced with RU486 in supernatant of the transfected HEK293 cells was measured by ELISA. Results The sequence of single chain mIL-12 what we obtained was the same as the expected result. The results of restriction endonuclease digestion and PCR showed that the RU486-inducible regulatory vector (pRS-RUmIL-12) was successfully constructed. No significant mIL-12 protein concentration in supernatant of HEK293 cells activation was measured without the inducer RU486, whereas higher concentration of the mIL-12 protein was observed in the presence of RU486. The relationship of concentration of the mIL-12 protein and RU486 was positive correlated under definite range. Conclusion A regulatable eukaryotic expression plasmid of mIL-12 single chain fusion gene was constructed, which could be used in the further research of gene regulation and gene therapy.
ObjectiveTo analyze the expression of VEGF, IL-33 and NO concentration after laser photocoagulation and subthreshold micropulse laser photocoagulation conventional in proliferative diabetic retinopathy (PDR) patients.MethodsA case control study. The clinical data of 39 patients of PDR and 11 patients of idiopathic macular pucker (IMP) from Department of Ophthalmology, Central Theater General Hospital during November 2015 were collected in this study. PDR patients were assigned randomly into three groups. Fifteen PDR patients with 15 eyes were treated with conventional laser as group A. Thirteen PDR patients with 13 eyes were treated with subthreshold micropulse laser as group B. Eleven PDR patients with 11 eyes without any laser therapy were grouped as C. Eleven IMP patients were grouped as D. There was no difference of age (F=0.53, P=0.23), gender ratio (χ2=0.55, P=0.91), body mass index (F=2.62, P=0.07), duration diabetes (F=0.29, P=0.75), glycoslated hemglobin (F=1.72, P=0.19) in four groups. All PDR patients were examined with FFA. Total protein was quantified by a bicinchoninic acid assay kit. Levels of VEGF, IL-33, NO were determined using enzyme-linked immunosorbent assay kits.ResultsThere was no difference of total protein in four groups (F=1.78, P=0.17). Group C had a higher VEGF level than group A and B (F=7.84, P=0.002). Group A had a higher IL-33 level than group C (t=4.15, P=0.02). There was no difference of IL-33 level in group B and C (t=1.34, P=0.20). Group D had a lower NO level than group A, B, C (F=38.42, P＜0.001). There was no difference of NO level in group A, B and C (F=3.29, P=0.06).ConclusionsBoth conventional laser photocoagulation and subthreshold micropulse laser photocoagulation can decrease vitreous VEGF level and subthreshold micropulse laser photocoagulation can induce less IL-33 level.
Objective To evaluate the role of interleukin-10 (IL-10) in acute pancreatitis. Methods Thirty mongrel dogs were divided into three groups based on the severity: acute edematous pancreatitis (AEP) group (n＝11), acute hemorrhagic necrotizing pancreatitis (AHNP) group (n＝12), and control group (n＝7). Serum level of IL-10 was determined with enzyme-linked immuno-sorbent assay (ELISA). Results Within 24 hours, AEP group had serum level of IL-10 significantly higher than that of AHNP group. Control group had no detectable serum IL-10. No significant difference was observed between AEP group and AHNP group at 48 hours. Conclusion The finding of low values of serum IL-10 suggests that there may be more consumption in AHNP group than in AEP group and it may be beneficial to decrease the severity of experimental acute pancreatitis.
Objective To investigate the expression and significance of Fork head /winged helix protein 3 (Foxp3) , retinoic acid-related orphan receptorγt (RORγt) , and interleukin-17 (IL-17) in Guinea pigs with emphysema. Methods Smoking and active immunization with elastin were separately used in guinea pigs to establish emphysema model. Then the destruction of lung tissue was assayed by measurement of the average radius of alveolar. The expressions of Foxp3 , RORγt, and IL-17 in lung tissue of the guinea pigs were detected by immunohistochemical technique. The results were compared with the normal control group by the analysis of variance or kruskal-Wallis test. Spearman rank correlation was used to analyze the correlation between the ratio of Foxp3/RORγt and IL-17, also the correlation between Foxp3/RORγt and the average radius of alveolar. Results In the smoking group and the active immunization group, the average radius of alveolar were significantly longer than the control group (Plt;0.05) . And the expression of Foxp3/RORγt was significantly unbalanced, with the number of Foxp3-positive cells decreased and RORγt-positive cells increased (Plt;0.05) . Meanwhile the level of IL-17 was significantly increased compared with the control group ( Plt;0.05) . The difference between the smoking group and the active immunization group was not significant (Pgt;0.05) . The ratio of Foxp3/RORγt was negatively correlated with the level of IL-17 and the average radius of alveolar. Conclusions Active immunization with elastin can induce emphysema in guinea pigs. The Foxp3/RORγt expression was unbalanced in lung tissue of guinea pigs with emphysema.This imbalance may be an important mechanism attributed to the disordered expression of CD4+ Treg cells and Th17 cells, which may be involved in autoimmune regulation and development of chronic obstructive pulmonary disease.