Objective To detect gene mutations of Fas gene death domain （exons 7-9） in 2 Chinese keloid pedigrees and to investigatethe significance of Fas gene mutations in the keloid formation.Methods The samples were selected from keloid pedigrees A and B in 2005. The polymerase chainreaction and DNA sequencing analysis technique were used to detect the sequenceof exons 7-9 of Fas gene from keloid tissues of 2 male patients in pedigree A,their peripheral vein blood and their surrounding normal skin served as their own contrast, their spouses’ peripheral vein blood served as normal contrast, the peripheral vein blood of 2 patients in pedigree B served as a contrast between different keloid pedigrees.Results No gene mutations and single nucleotidepolymorphism in Fas gene exons 7, 8 were found in all samples from pedigrees A and B. But point mutations and single nucleotide polymorphism in Fas gene exon 9were identified in 11 bp and 53 bpin 2 keloid tissue samples from Chinese keloid pedigree A.Conclusion Fas gene point mutations maybe indicate some relations in Fas protein function and keloid formation.
Objective To research the effect of γ-radiation released from 103Pd radioactive stent on the expression of Fas gene and its relation with apoptosis of bile duct cancer cells lines. Methods The cancer cells of bile duct were dissociated into suspension in culture flasks, and the number of cells was counted by hemacytometry. The suspension was then stored in 2 ml freezing tubes in the density of 1×105/ml. They were set in two stents: general stent （general stent group） and 103Pd radioactive stent (103Pd stent group). The expression of Fas gene and apoptosis of bile duct cancer cells in general stent group and 103Pd stent group were analysed with immunohistochemistry technique and TUNEL method, respectively. Results The expression level of Fas gene in 103Pd stent group was significantly higher than that in general stent group （P＜0.05）, and the number of apoptotic cancer cells in 103Pd stent group was also significantly more than that in general stent group （P＜0.01）. Conclusion There is a correlation between the expression of Fas gene and the apoptosis of bile duct cancer cells, which means that 103Pd radioactive stent may increase the expression of Fas gene and promote the apoptosis of cancer cells. It may be helpful for the further study of treatment for bile duct cancer using 103Pd radiative stent.
Objective To investigate the rationale of immune privilege of testicular sertoli cell. Methods Testicular sertoli cell was prepared by digested collagenase, trypsin, and Dnase. In vitro, the sertoli cells were culture together with active lymphocytes to observe the effect on killing lymphocytes. SABC was used for labeling the Fas ligand on testicular sertoli cell.Results In vitro, sertoli cell can kill the active lymphocytes, and testicular sertoli cell expresses the Fas ligand. Conclusion Fas ligand expressing on the testicular sertoli cell may be the cause of immune privilege of testicular.
【Abstract】ObjectiveTo measure the expressions of Fas/FasL mRNA in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma, and to explore the relationship between the expressions of Fas/FasL mRNA in those tissues and the hepatocellular carcinogenesis. MethodsSemi-quantity reverse transcript-ploymerase chain reaction(QRTPCR) were performed to measure the relative quantity of the Fas and FasL mRNA expressions in normal liver (n=25), adjacent noncancerous liver parenchyma(n=40) and hepatocarcinoma(n=40). ResultsThe relative quantity of Fas and FasL mRNA expressed in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma were 0.792±0.039 vs 0.245±0.043，0.857±0.031 vs 0.429±0.035 and 0.473±0.047 vs 0.185±0.041, respectively. The relative quantity of Fas mRNA expression in hepatocarcinoma was lower than that of normal liver tissue and adjacent non-cancerous liver parenchyrna (P＜0.05). The relative quantity of FasL mRNA expression in hepatocarcinoma was also lower than that of normal liver tissue (P＜0.05) and adjacent non-cancerous liver parenchyma (P＜0.01), but its expression in adjacent non-cancerous liver parenchyma was higher than that of normal liver tissue (P＜0.05).ConclusionHepatorcarcinoma may escape the immune surveillance of the host, not only by means of reducing Fas expression, but also through adjacent non-cancerous liver parenchyma’s increasing expression of FasL to induce apoptosis of contact lymphocyte which highly expresses Fas.
ObjectiveTo evaluate the effect of pre-infusion of allogeneic lymphoyctes treated with 5-FU on the rat liver graft. MethodsRat liver transplant models from Wistar to SD were established. Four groups were designed as following: control group: only liver transplantation without any other intervention; lymphocytes group: 1 ml of untreated lymphocytes (5×106/ml) from Wistar rats were preinfused into SD rats on day 7 and 4 separately before transplantation; lymphocytes with low concentration of 5-FU group: low concentration 5-FU (7.5 μg) treated lymphocytes were preinfused as above; lymphocytes with high concentration of 5-FU group: high concentration 5-FU (15 μg) treated lymphocytes were preinfused as above. Fas-L and CD8 expression were detected by immunohistochemistry method on day 7 after transplantation. ResultsThe integral opticaldensity (IOD) of Fas-L positive lymphocytes in the lobules of liver and portal areas were higher in lymphocytes with low concentration of 5-FU group than in the other groups （Plt;0.05）. There was no difference between lymphocyte group and lymphocytes with high concentration of 5-FU group （Pgt;0.05）. The IOD of CD8+ expression in lobules of liver was not different among all the three lymphocytes treated groups （Pgt;0.05）. But in portal areas, CD8+ expression was lower in the lymphocytes with low concentration of 5-FU group than in the other groups （Plt;0.05）. ConclusionPreinfusion of lymphocytes treated with low concentration 5-FU can induce graft immune tolerance, the probable mecanism of which is the increasing Fas-L expression in graft.
Objective To explore the expressions of Galectin-3, Fascin-1, and β-catenin protein in colorectal adenocarcinoma and the relations to clinicopathologic characteristics. Methods The expressions of Galectin-3, Fascin-1, and β-catenin protein were detected in 60 cases of colorectal adenocarcinoma, 30 cases of adenoma, and 30 cases of normal mucosa by microwave-EliVisionTM immunohistochemistry method, and analyzed the expressions of them and the relations to clinicopathologic characteristics. Results The expression rate of Galectin-3, Fascin-1, and β-catenin protein in CRC was 68.3% （41/60）, 53.3% （32/60）, and 81.7% （49/60） respectively, which was 46.7% （14/30）, 30.0% （9/30）, and 43.3% （13/30） respectively in adenoma, and 20.0% （6/30）, 3.3% （1/30）, and 13.3% （4/30） respectively in normal mucosa, the differences had statistical significance （P＜0.05）. The expressions of Galectin-3, Fascin-1, and β-catenin protein had statistically significant correlation with the TNM stage, invasive degree, and lymph node metastasis of colorectal adenocarcinoma （P＜0.05, P＜0.01）. The expressions of Galectin-3 and β-catenin protein had statistically significant correlation with the different differentiation degree of colorectal adenocarcinoma （P＜0.05）, but the expression of Fascin-1 protein was not related to differentiation degree of colorectal adenocarcinoma （P＞0.05）.The expressions of Galectin-3, Fascin-1, and β-catenin protein had not statistically significant correlation with the patient’s age and gender, and tumour size （P＞0.05）.There were positive correlations between the Galectin-3 and Fascin-1 or β-catenin （r=0.728，P＜0.01；r=0.696，P＜0.01）, and there was positive correlation between β-catenin and Fascin-1 （r=0.507，P＜0.01）. Conclusions The high expressions of Galectin-3, Fascin-1, and β-catenin protein in colorectal adenocarcinoma tissues are some extent correlated to the high invasive ability and lymph node metastasis, which could be used for the indexes to predict the invasion and metastasis in colorectal carcinoma potentially.
Objective To investigate the plasma levels of soluble Fas receptor ( sFas) , soluble Fas ligand ( sFas-L) and matrix metalloproteinase-7 ( MMP-7) and their correlation with disease severity as well as the prognosis of septic patients.Methods The plasma levels of sFas, sFas-L, sFas / sFas-L ratio and MMP-7 were measured by enzyme-linked immunosorbent assay and compared between32 patients with sepsis and 24 age and sex matched healthy controls. Based on the 28-day outcome, the patients were divided into a survival group and a death group. The difference in sFas, sFas-L, sFas/ sFas-L ratio and MMP-7 between the survival group and the death group were compared.Results Compared with the healthy control group, the concentration of plasma sFas, sFas-L and MMP-7 were significantly increased in the septic patients ( P lt; 0. 01) . Elevated plasma sFas and sFas-L were both positive correlated with the APACHEⅡ score and SOFA score. Although a modest negative correlation was found between plasma MMP-7 and APACHEⅡ score and SOFA score, but this correlation did not reach statistical significance ( P gt;0. 05) . The septic patients who died had significantly higher sFas-L level and lower sFas / sFas-L ratio as compared with those who survived ( P lt;0. 05) . Conclusion Plasma sFas, sFas-L and MMP-7 are associated with the disease severity and can serve as potential markers for predicting the outcome in septic patients.
Objective To investigate the protective effects of recombinant human insulin-like growth factor-1 ( rhIGF-1) on apoptosis of diaphragm in rats with COPD and its impact on pulmonary function. Methods Forty-five male Wistar rats were randomly divided into three groups, ie. a normal control group, a model group, and an IGF-1 intervention group, with 15 rats in each group. The rats in the model group and IGF-1 group were exposed to 5% smoke ( 30 min perday, lasting 28 days) in a sealed box, and 200 μg lipopolysaccharide was injected intratracheally on the 1st and 14th day. The rats in the IGF-1 group were given rhIGF-1 ( 60 μg /100 g) additionally by subcutaneous injection once a day, lasting 28 days. On the 1st, 14th, 28th day, 5 rats from each group were sacrificed. The weight, rate of apoptosis, Fas gene and Fas protein expression of isolated diaphragms were detected. The pulmonary function was measured on the 28th day before sacrificed. Results The mass of diaphragms, minute ventilation ( VE) , peak expiratory flow ( PEF) , inspiratory capacity ( IC) , forced expiratory volume in 0. 3 second ( FEV0. 3) of themodel groupand IGF-1 group were all decreased compared with the control group ( P lt; 0. 05) . The mass of diaphragms, VE, IC of the IGF-1 group were higher than those of the model group ( P lt;0. 05) , and the differences of PEF and FEV0. 3 were not significant ( P gt; 0. 05) . On the 14th, 28th day, rate of apoptosis, Fas gene and protein expressions in the IGF-1 group were lower than those in the model group, and still higher than those in the control group ( P lt; 0. 05) . Conclusions Fas/FasL mediated apoptosis way is involved in the diaphragm apoptosis. rhIGF-1 may reduce the apoptosis of the diaphragmand improve the VE and IC of rats with COPD by intervening Fas/FasL pathway.
Objective To understand the molecular mechanism of HBx in the carcinogenesis of hepatitis B virus (HBV) related hepatocellular carcinoma (HCC).Methods The literatures published in the past 5 years which are mainly about HBx and hepatocellular carcinoma were reviewed. Results HBx had many functions, such as cell malignant transformation, inhibiting DNA repair, trans-activation, inhibiting p53 and apoptosis. These functions together with its Fas/Fas-L interfering and caspase-3 inhibiting could contribute to the carcinogenesis and development of HBV relatde HCC. Conclusion HBx has broad spectrum of biological functions, which contribute to the carcinogenesis and development of HBV related HCC.
Objective To investigate the effects of FasL gene-modified dendritic cell (DC) on the airway inflammation in mice sensitized/challenged by house dust mite (HDM) allergen.Methods FasL gene-modified DC (FasL-DC) and control DC (nontransfection DC) were administrated into HDM sensitized and challenged mice by intratracheal injection respectively，then HDM sensitized and challenged mice were sacreificed two days later.Total and differentiation cell counts and levels of interleukin-4（IL-4），IL-5 and interferon-γ(IFN-?) in bronchoalveolar lavage fluid (BALF) were detected and lung histological features were observed.Results After administration of FasL-DC，lung allergic inflammation was ameliorated while total cell counts，the percentage of eosinophil ,the levels of IL-4 and IL-5 in BALF decreased and the level of IFN-? in BALF increased.Conclusion Administrating FasL-DC into HDM sensitized/challenged mice can inhibit Th2 cells activation and ameliorate airway allergic inflammation.