1.Immunotolerance following intermingled skin transplantation
Chinese Journal of Tissue Engineering Research 2007;0(18):-
PUBMED Database and Vip Database were undertaken to identify relevant articles on intermingled skin transplantation. Skin cuticular layer consists of keratinocytes and dendritic cells (DCs), so the researches on intermingled skin transplantation includes the chimerism of allo-keratinocyte and the chimerism of allo-DCs. There is no study about the chimerism of allo-DCs at present. The investigations on skin microchimerism are limited to the immunotolerance induced by exogenous stem cell transplantation, and the microchimerism phenomenon has not been found in allo-skin transplantation and intermingled skin transplantation. Regulating T cells, Th3 cells and NKT cells play important roles in the induction of the immunotolerance after intermingled skin transplantation.
2.Repair and reconstruction mechanism of stromal cell derived factor-1 on rat abdominal aorta grafts
Chengjin GAO ; Wei XIONG ; Shishu HUANG
Chinese Journal of Organ Transplantation 2003;0(06):-
Objective To investigate the repair and reconstruction mechanism of stromal cell derived factor-1(SDF-1) on rat abdominal aorta grafts.Methods Male Sprague-Darley rats received abdominal aorta grafts from male Wistar abdominal arteries.Immunohistochemical staining(IHC) was performed to detect the SDF-1 expression.The expression of CXCR4,the receptor of SDF-1,was demonstrated by RT-PCR.(Results) SDF-1 was expressed on the endothelium all the time during the course of the abdominal aorta grafts in the bodies of recipients.Furthermore,it was found that there was a significant relationship between the expression of SDF-1 on the endothelia and the thickness of the neointima of the rat abdominal aorta grafts.On the other hand,CXCR4 was detected within the abdominal aorta grafts.Conclusions Stem cells could be mobilized by SDF-1 to the abdominal aorta grafts and differentiated into smooth-muscle like cells.Furthermore,SDF-1 is predictive of grafts arteriosclerosis and the subsequent development of chronic graft dysfunction(CGD).
3.The mechanism of SW480 cell line death caused by transfection of G1CEACDNa gene
Xiaojun WANG ; Qingjiu MA ; Danian LAI ; Chengjin LI ; Jianguo LU ; Jinmao LI ; Yan DENG ; Deming GAO
Chinese Journal of General Surgery 2000;0(11):-
Objective To study the mechanism of SW480 cell line death caused by transfection of retroviral vector-G1CEACDNa.Methods Plasmid G1CEACDNa was transferred into the SW480 cell line using liposomes method. RT-PCR was performed to examin the expression of CD gene indrectly.The cell growth curve was measured by means of cell counting. When the CD+SW480 cells were exposed to 5-FC (1mmol/L), the growth inhibition rate and the "bystander effect" were detected by MTT method.The ultrastructure was observed by electron microscope.Apoptosis was verified by flow cytometer .Results The product of RT-PCR showed a band at 1.5kb on the photo of electrophoresis. The growth of CD+ SW480 cells was inhibited 24h after administrating 5-FC,and the inhibition rates at 72h,96h,120h were 30.0%,50.0% and 80.0%,respectively.Apoptosis cells in different phases and apoptotic bodies in the field of electron microscope were observed. Meanwhile ,a few cells showed necrosis.Flow cytometer verified that a few cells appearred apoptosis 48h after exposed to 5-FC (1mmol/L), the apoptosis rate and the necrosis rate at 72h,96h were 20.2%,30.7% and 19.6%,21.1% respectively.Conclusions The death mechanism of SW480 cells transfected with G1CEACDNa followed by 5-FC treatment includes both necrosis and apoptosis.Apoptosis is possibly the bystander effect.
4.Detection of the effect of bortezomib with arsenic trioxide different concentration on the cell cycle and apoptosis of Raji cells by flow cytometry
Xiujuan ZHAO ; Yuliang SHI ; Ling XIAO ; Chengjin YAN ; Lihua MENG ; Zhilin GAO ; Hongyan JIN ; Xinlei GUO
Journal of Leukemia & Lymphoma 2010;19(12):721-723
Objective To investigate the effect of bortezomib with arsenic trioxide different concentration on the cell cycle and apoptosis of Raji cells. Methods Flow cytometry analysis showed that the relative number of cells in different phases and the percentages of cells calculated in G1 and S phase of the cell cycle and apoptosis were assessed after treatment with As2O3 and BOR or in combination with BOR in different concentration at indicated time (24 h, 48 h, 72 h). Results Flow cytometric analysis showed that the cell cycle was arrested at G1 phase, the number of cells G1 period increased significantly, and S phase decreased on Raji cells after As2O3 treatment. The relationship between the cellular DNA contents and the concentration of As2O3 showed a dose-and time-dependent manner (P <0.0001). But it was found that BOR had no effect on Raji cell cycle, but, in two drugs combination, cell apoptosis rate significantly increased from 16.98 % to 45.84 %. Conclusion The results show that As2O3 exerted variable and definite effects on lymphoma Raji cells, which indicated that As2O3 might induce apoptosis and arrest cell cycle. The combination of two drugs had a effective and synergistic effect on apoptosis.
5.Effect of thermochemotherapy on human colon cancer cell line SW480 transfected with cytosine deaminase gene
Jinmao LI ; Qingjiu MA ; Danian LAI ; Chengjin LI ; Jiangguo LU ; Xiaojun WANG ; Deming GAO
Chinese Journal of General Surgery 1997;0(06):-
Objective To investigate the effect of thermochemotherapy with 5 fluorocytosine(5 FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue specific cytosine deaminase (CD) in vitro.Methods Recombinant retroviral vector G1CEACDNa, in which CD gene was controlled under the CEA promoter, was introduced through liposome technique to the human colorectal carcinoma cell line SW480, and then the cells were selectively cultured in G418. The proliferated colonies were treated with the combined therapy of 5 FC and hyperthermia at a 43℃ over 30 min for 3 times. RT PCR was performed to analyze the expression of CD gene in target cells after thermochemotherapy. Results The CD gene expressed steadily in the target cells after 3 times of hyperthermic treatment. After the transfection of CD gene, SW480 CEACD cells were more sensitive to 5 FC than their parental cells (P
6.Changes in circulating microparticles in mice with ventilator-induced lung injury
Yingjie YIN ; Shouqin ZHANG ; Sanqiang WANG ; Hu LONG ; Chengjin GAO ; Xiangyu ZHANG
Chinese Critical Care Medicine 2018;30(1):8-12
Objective To investigate the changes in serum level of microparticles (EMPs) in mice with ventilator-induced lung injury (VILI), and explore its significance in VILI. Methods Forty-eight grade SPF male C57BL/6J mice were randomly divided into two groups, with 24 mice in each group: the mice in mechanical ventilation (MV) group were given high tidal volume (VT 30 mL/kg) MV for 4 hours after tracheal intubation, and those in spontaneous breathing group were spontaneously breathed for 4 hours. The apical blood of 12 mice in each group were collected, and serum levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA), and serum EMPs levels were determined by flow cytometer. The correlations between EMPs and IL-1β, IL-6, and TNF-α were analyzed by linear regression analysis. The lung tissues of other 12 mice in each group were harvested, and wet/dry weight (W/D) ratio was assessed. After hematoxylin-eosin (HE) staining, the morphological changes in lung tissue were observed under light microscope. After double staining of uranium acetate and lead citrate, the ultrastructural changes in lung tissue were observed with electron microscope. Results Compared with spontaneous breathing group, the levels of lung W/D ratio in MV group was significantly increased (5.47±0.14 vs. 4.34±0.11), the levels of IL-1β, IL-6, TNF-α and EMPs were also significantly increased [IL-1β (ng/L): 42.4±4.4 vs. 7.7±3.6, IL-6 (ng/L): 1 239.5±66.3 vs. 21.7±4.6, TNF-α (ng/L):237.6±25.8 vs. 37.1±19.1, EMPs (cells/μL): 28.6±1.8 vs. 5.9±1.8, all P < 0.01]. It was shown by correlation analysis that EMPs were positively related with IL-1β, IL-6, and TNF-α (r value was 0.968, 0.932, 0.945, respectively, all P = 0.000). It was shown by fitting linear regression analysis that when EMPs increased by 1 cell/μL, IL-1β increased by 2.4 ng/L [95% confidence interval (95%CI) = 1.9-2.8, P < 0.001], IL-6 increased by 34.5 ng/L (95%CI = 25.1-44.0, P < 0.001), and TNF-α increased by 13.6 ng/L (95%CI = 10.3-16.9,P < 0.001). It was shown by light microscope that the structure of lung tissue and alveolar of mice in spontaneous breathing group appeared normal, while the shrinks of alveolar and disappearance of alveolar architecture were found in MV group. It was shown by electron microscopy that alveolar wall edema and thickening and broken alveolar septa were found in MV group, by contrast, the structure of alveolar was normal in spontaneous breathing group. Conclusion 30 mL/kg VT ventilation for 4 hours could induce VILI with increase in EMPs, suggesting EMPs closely related to VILI, and EMPs level may be putative biomarker of VILI.
7.PAI-1 genetic polymorphisms influence septic patients' outcomes by regulating neutrophil activity.
Shaowei JIANG ; Yang WANG ; Liang CHEN ; Honghua MU ; Connor MEANEY ; Yiwen FAN ; Janesh PILLAY ; Hairong WANG ; Jincheng ZHANG ; Shuming PAN ; Chengjin GAO
Chinese Medical Journal 2023;136(16):1959-1966
BACKGROUND:
Plasminogen activator inhibitor-1 (PAI-1) plays an important role in the pathophysiology of sepsis, but the exact mechanism remains debatable. In this study, we investigated the associations among the serum levels of PAI-1, the incidence of 4G/5G promoter PAI-1 gene polymorphisms, immunological indicators, and clinical outcomes in septic patients.
METHODS:
A total of 181 patients aged 18-80 years with sepsis between November 2016 and August 2018 in the intensive care unit in the Xinhua Hospital were recruited in this retrospective study, with 28-day mortality as the primary outcome. The initial serum level of PAI-1 and the presence of rs1799768 single nucleotide polymorphisms (SNPs) were examined. Univariate logistic regression and multivariate analyses were performed to determine the factors associated with different genotypes of PAI-1, serum level of PAI-1, and 28-day mortality.
RESULTS:
The logistic analysis suggested that a high serum level of PAI-1 was associated with the rs1799768 SNP of PAI-1 (4G/4G and 4G/5G) (Odds ratio [OR]: 2.49; 95% confidence interval [CI]: 1.09, 5.68). Furthermore, a high serum level of PAI-1 strongly influenced 28-day mortality (OR 3.36; 95% CI 1.51, 7.49). The expression and activation of neutrophils (OR 0.96; 95% CI 0.93, 0.99), as well as the changes in the expression patterns of cytokines and chemokine-associated neutrophils (OR: 1.00; 95% CI: 1.00, 1.00), were both regulated by the genotype of PAI-1.
CONCLUSIONS
Genetic polymorphisms of PAI-1 can influence the serum levels of PAI-1, which might contribute to mortality by affecting neutrophil activity. Thus, patients with severe sepsis might clinically benefit from enhanced neutrophil clearance and the resolution of inflammation via the regulation of PAI-1 expression and activity.
Adolescent
;
Adult
;
Aged
;
Aged, 80 and over
;
Humans
;
Middle Aged
;
Young Adult
;
Genotype
;
Neutrophils
;
Plasminogen Activator Inhibitor 1/genetics*
;
Polymorphism, Single Nucleotide/genetics*
;
Retrospective Studies
;
Sepsis/genetics*