1.Effect of high mobility group box-1 protein on Foxp3 expression in spleen regulatory T cells in mice
Ying ZHANG ; Yongming YAO ; Ning DONG ; Feng LIU ; Zhiyong SHENG
Journal of Third Military Medical University 2003;0(21):-
Objective Intranuclear forkhead/winged helix transcription factor p3(Foxp3) plays a key role in T cell-mediated immunosuppression.The present study was performed to investigate the effects of high mobility group box-1 protein(HMGB1) on Foxp3 gene as well as protein expressions in splenic regulatory T cells(Tregs) and their potential regulating mechanisms in mice.Methods CD4+CD25+Tregs isolated from the spleens of male BABL/c mice by magnetic beads were seeded on 96-well(1?105 cells/well) cell culture plates coated with anti-CD3(1 ?g/ml) and soluble anti-CD28(1 ?g/ml),and the cells were stimulated with HMGB1 at various intervals or at different concentrations.After being stimulated,the Foxp3 mRNA/protein expressions in the Tregs were determined.The time-dependent and dose-dependent responses between HMGB1 and intranuclear Foxp3 expression were analyzed by flow cytometry,and the expressions of Foxp3 mRNA of Tregs were analyzed by quantitative PCR of SYBR GREEN.Results After stimulation with HMGB1,the intranuclear Foxp3 protein and mRNA expressions of splenic Tregs in mice were markedly down-regulated in 24 h to 72 h(P
2.Four-tube strategy for the treatment of thoracic cavity fistula following esophagus carcinoma resection
Sheng YAO ; Canhui LIU ; Kang WANG ; Demin LI ; Guohua DONG
Journal of Medical Postgraduates 2016;29(8):845-848
Objective Thoracic cavity fistula following esophagus carcinoma resection is a serious complication with a high mortality.This study aims at a better therapy for thoracic cavity fistula following esophagus carcinoma resection by summarizing the ex-perience with the four-tube strategy ( jejunal fistula tube, stomach tube, chest drainage tube, and nasal fistula tube) in the treatment of the complication. Methods We retrospectively analyzed the clinical data about 62 cases of thoracic cavity fistula following esopha-gus carcinoma resection, 35 treated with the four-tube strategy ( treatment group) and the other 27 with the three-tube ( stomach tube, chest drainage tube, and nasal fistula tube) method ( control group) .We compared the hospital days, wound healing time, mortality, and incidence of anastomotic stenosis at 6 months after operation between the two groups of patients. Results Compared with the controls, the treatment group showed remarkable decreases in the hospital days (P<0.05), wound healing time (P<0.05), and mortality (P<0.05), but no statistically significant difference was observed in the incidence rate of anastomotic stenosis at 6 months after operation between the two groups of patients ( P >0.05 ) . Conclusion Compared with the three-tube method, the four-tube strategy has the advantages of shorter healing time and lower mortali-ty, and therefore is preferable for the treatment of thoracic cavity fis-tula following esophagus carcinoma resection.
3.The Assembly of a Novel Enzyme Biosensor for Aflatoxin B_1 Detection
Da-Ling LIU ; Yi SHEN ; Jing ZHANG ; Dong-Sheng YAO ;
China Biotechnology 2006;0(03):-
A novel biosensor for aflatoxin B1 detecting has been reported. The biosensor electrode for AFB1 detecting was assembled by immobilized aflatoxin-oxidoreductase using open-ended multi-walled carbon nanotubes as matrix. Its linear range was between 0.16?M and 3.2?M. And if the specific anti-aflatoxin B1 antibody and aflatoxin oxidoreductase were both immobilized on the electrode with Multi-Walled carbon nanotubes, the detection limit of the modified electrode could be 16 nM with a 10 times improved sensitivity. The aflatoxin enzyme biosensor assembled this way strode one step forward its practical application.
4.Changes and significance of cytokine signaling suppressor mRNA in burn rats with Staphylococcus aureus sepsis
Hongyun LI ; Yongming YAO ; Fenghua YAO ; Ning DONG ; Yan YU ; Zhiguo SHI ; Zhiyong SHENG
Chinese Journal of Trauma 2003;0(12):-
Objective To observe the expression law of cytokine signaling suppressors (SOCSs) mRNA in burn rats with Staphylococcus aureus sepsis and investigate their potential role in the pathogenesis of postburn sepsis. Methods Wistar rats were inflicted with 20% TBSA Ⅲ? scald, followed by Staphylococcus aureus challenge. Then, the expressions of SOCS1, SOCS2 and SOCS3 mRNA and interferon-? (IFN-?) levels in the liver and lungs were determined. Results With Staphylococcus aureus challenge after burn, IFN-? levels in the liver and lungs were significantly elevated and reached peak at the 0.5th and 6th hours, respectively (P
5.TNF-α and plasma D(-)-lactate levels in rats after intestinal ischemia and reperfusion
Yongming YAO ; Ailan REN ; Shengli DONG ; Ning DONG ; Yan YU ; Zhiyong SHENG
Journal of Geriatric Cardiology 2004;1(2):119-124
Objective To study the potential role of tumor necrosis factor-α (TNF-α) induction in the development of mucosal barrier dysfunction in rats caused by acute intestinal ischemia-reperfusion injury, and to examine whether pretreatment with monoclonal antibody against TNF-α (TNF-α MoAb) would affect the release of D(-)-lactate after local gut ischemia followed by reperfusion. Methods Anesthetized Sprague-Dawley rats underwent superior mesenteric artery occlusion for 75 min followed by reperfusion for 6 hr. The rats were treated intravenously with either TNF-α MoAb (20 mg/kg) or albumin (20 mg/kg) 30 min prior to the onset of ischemia. Plasma D(-)-lactate levels were measured in both the portal and systemic blood by an enzymatic spectrophotometric assay. Intestinal TNF-αmRNA expression as well as protein levels were also measured at various intervals. In addition, a postmortem examination was performed together with a macropathological evaluation based on a four-grade scoring system.Results Intestinal ischemia resulted in a significant elevation in D(-)-lactate levels in the portal vein blood in both the control and treatment groups ( P <0.05). However, animals pretreated with TNF-α MoAb at 6 hr after reperfusion showed significant attenuation of an increase in both portal and systemic D(-)-lactate levels when compared with those only receiving albumin (P < 0.05). In the control animals, a remarkable rise in intestinal TNF-α level was measured at 0.5 hr after clamp release ( P < 0.01); however, prophylactic treatment with TNF-α MoAb completely annulled the increase of local TNF-α levels seen in the control animals. Similarly, after anti-TNF-α MoAb administration, intestinal TNF-α mRNA expression was markedly inhibited, which showed significant differences when compared with the control group at 0.5 hr, 2 hr and 6 hr after the release of occlusion ( P < 0.05-0.01 ). In addition, the pathological examination showed marked intestinal lesions that formed during ischemia, which were much worse upon reperfusion,particularly at the 6 hr time point. These acute injuries were obviously attenuated in animals receiving TNF-α MoAb.Conclusions It appeared that acute intestinal ischemia was associated with failure of the mucosal barrier, resulting in increased plasma D(-)-lactate levels in both portal and systemic blood. These results suggest that TNF-α appears to be involved in the development of local damage associated with intestinal ischemic injury. Moreover, prophylactic treatment with TNF-α MoAb exerts preventive effects on ischemia/ reperfusion-induced circulating D (-)-lactate elevation and gut injury. ( J Geriatr Cardiol 2004;1(2):119-124. )
6.Immunological activity change of regulatory T cells and its relation with outcome of patients with severe burn
Lifeng HUANG ; Yongming YAO ; Ning DONG ; Yan YU ; Lixin HE ; Zhiyong SHENG
Chinese Journal of Trauma 2010;26(9):785-789
Objective To investigate the immunological activity change of regulatory T cells (Treg) and discuss its significance in the outcomes of patients with multiple organ dysfunction syndrome (MODS) and severe burn. Methods A total of 106 patients with total burn surface area (TBSA) larger than 30% were included in the study and randomly divided into three groups according to the burn area: Group Ⅰ (TBSA of 30%-49%, n = 41), Group Ⅱ (TBSA of 50% -69%, n = 34) and Group Ⅲ (TBSA of 70%-99%, n = 31). According to the development of MODS, patients were divided into MODS group (n =21) and non-MODS group (n =85). The patients with MODS were further divided into non-survival group (n = 16) and survival group (n = 5) based on their outcomes. Healthy volunteers were served as normal control (n = 25). Peripheral blood samples were collected at days 1,3,7, 14 and 21 postburn. The immunomagnetic separation technique was applied to separate and purify CD4+ CD25+Tregs in peripheral blood, and phenotypes (CTLA-4) were analyzed by flow cytometry and the contents of interleukin-10 released in the supernatants were determined by ELISA. Results Expression of CTLA-4 and level of IL-10 were significantly increased in burn patients compared with normal control group, with statistical differences. The expression of CTLA-4 and level of IL-10 were significantly increased in patients with severe burns at all time points. The expression of CTLA-4 and level of IL-10 in MODS group were much higher than those in non-MODS group at days 3-21 postburn (P < 0.01). Among the MODS patients, the expression of CTLA-4 and level of IL-10 in the survival group were obviously lower than those in the non-survival group at days 3-21 postburn (P < 0.05 or P < 0.01). Conclusions After severe burn injury, expressions of the markers on CD4 + CD25 + Treg surface and secretion of cytokines produced by CD4 + CD25 + Tregs show significant difference in patients with different born areas, MODS development and survival state. CD4 + CD25 + Treg may play an important role in the pathogenesis of immunoregulation, MODS and mortality of burn patients through secretion of inhibitory cytokines.
7.Application of three-dimensional technology in assessment of burn area and treatment strategy of burns.
Wen-bo SHENG ; Guosheng DONG ; Yan WAN ; Li YAO ; Hongtai TANG ; Zhaofan XIA
Chinese Journal of Burns 2014;30(4):353-355
Accurate area assessment of a burn injury and its treatment according to its depth of injury are the foundation of burn treatment due to its complexity, and various techniques and methods have been employed to solve these problems for many years. As the demand of modern medicine calls for individualized and precise therapeutic measures, it is clear that the traditional diagnostic and treatment measures are insufficient. The flourishing development of three-dimensional (3D) technology seems to provide new research approaches and technical opporturities for burn surgery. A series of techniques such as 3D model, 3D scanning, and 3D printing may be promising in advancing burn surgery through basic research to achieve rational clinical applications in the future. In this paper, the applications and achievements of 3D technology in burn surgery in recent years are summarized.
Body Surface Area
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Burns
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diagnosis
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pathology
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therapy
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Humans
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Image Processing, Computer-Assisted
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methods
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Therapy, Computer-Assisted
8.The potential role of staphylococcal enterotoxin B in the early intestinal injury in postburn Staphylococcus aureus sepsis
Hongyun LI ; Yongming YAO ; Zhiguo SHI ; Ning DONG ; Yan YU ; Lianrong LU ; Zhiyong SHENG
Chinese Journal of Pathophysiology 1986;0(04):-
AIM: To investigate the role of staphylococcal enterotoxin B (SEB) in early intestinal injury in scald rats with Staphylococcus aureus sepsis. METHODS: 86 male Wistar rats were randomly divided into four groups as follows: normal controls ( n= 10), scald control group( n= 10), postburn sepsis group( n= 50) and SEB monoclonal antibody (MAb)treatment group ( n= 16). Plasma samples were collected to determine SEB, endotoxin, tumor necrosis factor-? (TNF-?) and interferon-? (IFN-?). RESULTS: After scald injury followed by Staphylococcus aureus challenge, the levels of SEB, TNF-? and IFN-? in plasma were significantly higher than those of normal controls, peaking at 2-6 h ( P
9.Prokaryotic expression of surface membrane antigen SAG1 gene from Toxoplasma Gondii and the diagnostic value of the recombinant protein
Chao-lan, WANG ; Dong-sheng, TANG ; Yong, YAO ; Xue-long, WANG ; Ye-mei, WANG
Chinese Journal of Endemiology 2011;30(4):376-378
Objective To investigate the diagnostic value of the recombinant surface antigen 1 (rSAG1) in immunodiagnosis of toxoplasmosis. Methods Isopropyl β-D- 1 -thio-galaetopyranoside (IPTG) was used to induce the expression of recombinant plasmid pET28a-SAG1 of Escherich coli(pET28a-SAG1/BL21 ). The expression products (rSAG1) of pET28a-SAG1/BL21 were identified by Western blotting. The serum of mice infected with Toxoplasma gondii tachyzoites, normal mouse serum and the serum from 10 toxoplasma gondii patients were used as primary anti-Toxoplasma gondii antibodies, and the rSAG1 gene products were identified by Western blotting, by which the diagnostic value of rSAG1 in Toxoplasmosis was compared. Results After induction and purification, rSAG1 protein was obtained and its relative molecular mass was 38.5 × 103. The fusion protein could be recognized by the serum of mouse infected with Toxoplasma gondii tachyzoites, rSAG1 of expression products of surface membrane antigen SAG1 gene from Toxoplasma Gondii could be detected in 4 cases from 10 patients by Westem blotting.Conclusion The rSAG1 has a potential value in the immunodiagnosis of Toxoplasmosis.
10.Study on the rehabilitation in post-stroke patients with shoulder-hand syndrome by manipulation treatment
Fu-sheng DONG ; Yu-qing DING ; Wen-hui ZHANG ; Shizhu YAO ; Minsheng WEN ; Weijin ZHOU
Chinese Journal of Rehabilitation Theory and Practice 2002;8(3):162-163
ObjectiveTo observe the rehabilitative effect of patients with shoulder-hand syndrome after stroke by manipulation treatment. MethodsThe patients with shoulder-hand syndrome were randomly divided into two groups, manipulation group (180cases) and control group (128 cases). Patients in the manipulation group were regularly given a passive quantitative movement on shoulder, elbow and hand joints,while patients in the control group were irregularly given a passive movement or ordered to perform an autonomic movement. The signs and symptoms of patients in these two groups were not much different. The rehabilitative effects were compared 3 months later. ResultsSigns and symptoms in the manipulation groups improved much better than that of the control group. Conclusions The manipulation treatment for the post-stroke patients with shoulder-hand syndrome is the method that is simple, effective and easy to perform.