MiR-125b inhibits growth of gastric cancer cells by targeting Fas/FasL signaling pathway / 中国医师杂志

Objective:To investigate the regulation effect of miR-125b in the gastric cancer cell growth mediated by apoptosis related protein (Fas)/apoptosis related protein ligand (FasL) signal.Methods:Gastric cancer SGC-7901 cells were cultured in vitro. MiR-125b inhibitor sequence, NC sequence and transfection reagent were transfected into SGC-7901 cells and divided into three groups: miR-125b inhibited group, NC group and control group. The expression of miR-125b in transfected cells was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), and cell proliferation was detected by cell counting kit-8 (CCK-8) method. The colony formation was detected by plate cell clone formation assay. Cell apoptosis and cycle were detected by flow cytometry. The protein expression of Fas and FasL was detected by Western blot. The targeted regulation of Fas by miR-125b was detected by luciferase activity assay. Results:The expression level of miR-125 and the number of cell colony in miR-125b inhibited group was significantly lower than those in control group and NC group, and the inhibition rate of cell proliferation and apoptosis rate were significantly higher than that in control group and NC group (all P<0.05). The DNA content in G 1 phase in miR-125b inhibited group was significantly higher than that in control group and NC group, and the DNA content in S phase in miR-125b inhibited group was significantly lower than that in control group and NC group (all P<0.05). The expression of Fas and FasL protein in miR-125b inhibited group was significantly higher than that in control group and NC group (all P<0.05). The target site of miR-125b was found in 3′-UTR of Fas mRNA, and compared with the NC+ Fas 3′UTR-Wt group, the activity of luciferase in the miR-125b inhibited group+ Fas 3′-UTR-Wt group decreased significantly ( P<0.05). Conclusions:Inhibition of miR-125b expression can activate Fas/FasL signal and inhibit SGC-7901 cell proliferation, induce G 1 phase arrest of cell cycle and promote apoptosis.

Anti-tumor effect and its related mechanisms of cinobufotalin combined with cisplatin on H22 liver cancer mice / 中国中药杂志

In order to observe the anti-tumor effect of cinobufotalin on H22 liver cancer mice and to explore its regulatory mechanism, 50 Kunming mice were subcutaneously inoculated with H22 intraperitoneal passage cells under the armpit to establish H22 hepatocellular carcinoma model. They were then randomly divided into model group, cinobufotalin low dose group, cinobufotalin high dose group, cisplatin group and cisplatin+cinobufotalin group, which received 0.01% ethanol solution, 1 mg·kg~(-1) cinobufotalin, 5 mg·kg~(-1) cinobufotalin, 5 mg·kg~(-1) cisplatin, 5 mg·kg~(-1)cisplatin + 5 mg·kg~(-1) cinobufotalin respectively for 10 days. The general condition of mice during the intervention was observed, and the inhibition rate, tumor mass, thymus index, histopathological changes of the tumors, apoptotic rate of the tumors, the expressions of phosphatidylinositol 3-kinase(PI3 K), protein kinase B(Akt), apoptosis related gene(Fas), Fas ligand(FasL) mRNA and protein phosphorylated Akt(pAkt) protein in the tumors of each group were compared. The results showed that during the modeling period, the mice showed a decline in food intake, dark fur, poor mental status, and gradually worsened over time. The mental status of mice in each intervention group was improved gradually, especially in the cisplatin+cinobufotalin group. As compared with the model group, the tumor mass of each intervention group was lower(P<0.05). As compared with the cinobufotalin low dose group, the tumor mass was lower and inhibition rate was higher in the cinobufotalin high dose group, cisplatin group and cisplatin+cinobufotalin group(P<0.05). As compared with the cinobufotalin high dose group and the cisplatin group, the tumor mass was lower and the inhibition rate was higher in cisplatin+cinobufotalin group(P<0.05). As compared with the model group, the thymus index was higher in cinobufotalin high dose group and cisplatin + cinobufotalin group, while was lower in cisplatin group(P<0.05). As compared with the cinobufotalin low dose group, the thymus index was higher in the cinobufotalin high dose group and lower in the cisplatin group(P<0.05). As compared with the cinobufotalin high dose group, the thymus index was lower in cisplatin group(P<0.05). As compared with cisplatin group, the thymus index was higher in cisplatin+cinobufotalin group(P<0.05). Pathological staining showed that a large number of heterogeneous cells and mitotic phenomena were observed in the model group. Cell fragments and neutrophils were observed in the tumor tissues of the intervention groups, showing diffuse necrosis, and the diffuse necrosis was more obvious in the cisplatin+cinobufotalin group. As compared with the model group, the apoptotic rate of the tumors and the relative expressions of Fas mRNA and protein were higher in the intervention groups, while the relative expressions of PI3 K, FasL mRNA and protein and the relative expression of pAkt protein were lower in the intervention groups(P<0.05). As compared with the cinobufotalin low dose group, the apoptotic rate of the tumors and relative expression of Fas and protein were higher in the cinobufotalin high dose group, cisplatin group and cisplatin+cinobufotalin group, while the relative expressions of PI3 K, FasL mRNA and protein and pAkt protein were lower(P<0.05). As compared with the cinobufotalin high dose group and the cisplatin group, apoptotic rate of the tumors and the relative expression of Fas mRNA and protein were higher in the cisplatin+cinobufotalin group, while the relative expressions of PI3 K, FasL mRNA and protein and pAkt protein were lower in the cisplatin+cinobufotalin group(P<0.05). In summary, cinobufotalin has significant anti-tumor effect on H22 liver cancer mice, and can enhance the immune function of mice and synergistically enhance the effect of chemotherapy. Its mechanism may be associated with regulating PI3 K/Akt/Fas/FasL signaling pathway related genes and protein expression.

Survival of isolated human preantral follicles after vitrification: Analyses of morphology and Fas ligand and caspase-3 mRNA expression / 대한생식의학회지

OBJECTIVE: This study aimed to examine the effect of vitrification on apoptosis and survival in human preantral follicles after thawing.METHODS: This experimental study was conducted at an acute tertiary care hospital from March 2012 to April 2013. Ovaries were sliced into 5×5×1-mm pieces and divided into the following three groups: preantral follicle isolation, ovarian tissue vitrification-warming followed by follicle isolation, and immunohistochemistry of fresh ovarian tissue. For statistical analyses, the Student t-test, chi-square test, Kruskal-Wallis test, and Kaplan-Meier survival analysis were used.RESULTS: A total of 161 preantral follicles (70% secondary) were collected from ovarian cortex tissue of six women between 30 and 37 years of age who underwent oophorectomy due to cervical cancer or breast cancer. There were no significant differences in the follicular morphology of fresh preantral follicles and vitrified follicles after thawing. The mean Fas ligand (FasL) mRNA expression level was 0.43±0.20 (relative to β-actin) in fresh preantral follicles versus 0.51±0.20 in vitrified follicles (p=0.22). The mean caspase-3 mRNA expression level in fresh preantral follicles was 0.56±0.49 vs. 0.27±0.21 in vitrified follicles (p=0.233). One vitrified-thawed secondary follicle grew and developed to an antral follicle within 6 days of culture.CONCLUSION: Vitrification did not affect preantral follicle morphology or mRNA expression of the apoptosis markers FasL and caspase-3. Further studies are required to establish whether vitrification affects the outcomes of in vitro culture and the maturation of preantral follicles.

Soluble Fas Ligand as a Potential Marker of Severity of Dengue Infection

Background: The apoptosis of microvascular endothelial cells causes plasma leakage in dengue haemorrhagic fever patients. The soluble Fas ligand is a protein with molecular weight of 40 kDa that acts as a mediator of apoptosis. This study aimed to prove whether soluble Fas ligand can be used as a potential marker to predict the severity of dengue infection by comparing the soluble Fas ligand levels in dengue fever (DF) and dengue haemorrhagic fever (DHF) patients early in the course of illness. Method: This was a prospective study. It included 42 dengue patients (22 DF patients and 20 DHF patients) and 20 healthy people as a control group. The soluble Fas ligand was measured by the enzyme-linked immunosorbent assay (ELISA). Result: Soluble Fas ligand was increased significantly (P < 0.001) in DHF patients (median = 130.19, IQR = 36.26) compared to DF patients (median = 104.73, IQR = 53.94) and the control group (median = 87.16, IQR = 24.91). Conclusion: Soluble Fas ligand can be used as a potential marker to predict the severity of dengue infection in the early course of the illness. However, a larger sample size and further objective studies are needed to confirm these findings.

Dynamic changes of FasL protein expression in retinal tissue with aging in retinal degeneration mice / 中华实验眼科杂志

Background Retinal transplantation is a new approach to the treatment of retinal degeneration diseases,and how to avoid or reduce the immune rejection after transplantation is a problem.In experimental studies on retinal transplantation,C57BL/6 mice are often used as donors and rd mice serve as recipients.Studies showed that Fas ligand (FasL) protein induces the apoptosis of Fas+ inflammatory cells by FasL/Fas signal pathway,speculating that FasL protein is associated with immune rejection after transplantation.Objective The aim of this study was to investigate FasL protein expression change in retinas of C57BL/6 mice and rd mice with aging and reveal the immune characteristics of the mouse retinas.Methods The frozen sections of eyeball from C57BL/6 mice and rd mice at the age of postnatal (PN)-0 week,PN-1 week,PN-2 week,PN-3 week and PN-4 week were prepared.The expression of FasL protein in the mouse retinas was examined by immnofluorescense technique.Images were acquired by fluorescence microscope and analyzed semi-quantitively by software from laser scanning confocal microscope as the fluorescence intensity (FI).The results were compared among different strains of mice.Results The retina developed imperfectly in PN-1 week C57BL/6 mice and FasL protein was positively expressed in the whole retina.In PN-2,3 and 4 week C57BL/6 mice,retinas finished the development with 10 layers,and retinal pigment epithelium (RPE),inner segment (IS),outer limiting membrane (OLM),outer plexiform layer (OPL),inner nuclear layer (INL),inner plexiform layer (IPL) and ganglion cell layer (GCL).Retinal structure and expression of FasL protein in the whole retina of rd mice in the PN-1 week were similar with C57BL/6 mice,however,ONL cells of rd mice were evidently decreased with aging.The RPE,OPL,INL,IPL and GCL expressed the FasL protein in PN-2,PN-3 and PN-4 week rd mice.The mean FI of FasL protein in the RPE layer was 184.199±16.747,186.797±7.904 and 184.319± 18.795 in rd mice of PN-2 week,PN-3 week and PN-4 week,which were significantly higher than 160.402±22.851,160.995 ±22.799 and 105.787 ± 17.676 in C57BL/6 mice (t =-3.360,P =0.002;t'=-4.277,P =0.000;t =-12.175,P=0.000).There were not significant differences in the mean FI of FasL protein in IPL between C57BL/6 mice and rd mice at ages of PN-2 week,PN-3 week and PN-4 week (all at P＞0.05).Conclusions The cells of retinal ONL are gradually decreased with the development of rd mice,and FasL protein expression intensity in RPE is evidently enhanced in rd mice compared with C57BL/6 mice.

Pentoxifylline and prostaglandin E1 action on ischemia and reperfusion of small intestine tissue in rats. An immunohistochemical study

PURPOSE: To investigate the action of pentoxifylline (PTX) and prostaglandin E1 (PGE1) on ischemia and reperfusion of small intestine tissue in rats, using immunohistochemical analysis. METHODS: Thirty-five Wistar rats were distributed as follows: group A (n=10): subjected to intestinal ischemia and reperfusion for 60 min, with no drugs; group B (n=10): PTX given during tissue ischemia and reperfusion; group C (n=10): PGE1 given during tissue ischemia and reperfusion; group D (n=5): sham. A segment of the small intestine was excised from each euthanized animal and subjected to immunohistochemical examination. RESULTS: Mean number of cells expressing anti-FAS ligand in the crypts was highest in Group A (78.9 ± 17.3), followed by groups B (16.7 ± 2.8), C (11.3 ± 1.8), and D (2.5 ± 0.9), with very significant differences between groups (p<0.0001). CONCLUSIONS: The use of pentoxifylline or prostaglandin E1 proved beneficial during tissue reperfusion. The immunohistochemical results demonstrated a decrease in apoptotic cells, while protecting other intestinal epithelium cells against death after reperfusion, allowing these cells to renew the epithelial tissue. .

Autoimmune lymphoproliferative syndrome:a case report and literature review / 北京大学学报(医学版)

SUMMARY We described 1 case of autoimmune lymphoproliferative syndrome ( ALPS) , first diagnosed in our hospital, and reviewed the recent literature. The 11-month old male patient presented with a histo-ry of splenomegaly and hepatomegaly since 1 month after birth. He suffered recurrent infectious diseases including cytomegalovirus infection, parvovirus B19 infection and chronic diarrhea disease. Besides, his symptoms included hemolytic anemia and thrombocytopenia. The laboratory abnormality indicated an ex-panded population of alpha/beta double-negative T cells (DNTs) (27. 18% of lymphocytes, 35. 16% of CD3 + T lymphocytes) in peripheral blood, and autoantibodies including antinuclear antibody, double-stranded DNA and rheumatic factor were positive. Hyper gamma globulinemia and positive direct Coombs tests were seen in the patient. His parents were both healthy and denied autoimmune diseases. We iden-tified a heterozygous point mutation in exon 3 of the FAS gene carrying c. 309 A>C, resulting in a single base pair substitution in exon 3 of FAS gene which changed the codon of Arg103 to Ser103 . Unfortunate-ly, we were unable to obtain the gene results of the child' s parents. The patient was treated with glu-cocorticoids in our hospital and with mycophenolatemofetil in other hospital. And we were informed that his anemia condition relieved through the telephone follow-up, but he still suffered recurrent infections, hepatomegaly and splenomegaly still existed. As we all know ALPS is characterized by defective lympho-cyte apoptosis, and thus cause lymphoproliferative disease and autoimmune disease, and increase the risk of lymphoma. It is more likely to be misdiagnosed as other diseases. ALPS should be suspected in the case of chronic lymphadenopathy, splenomegaly and autoimmune features. Flow cytometry approach is helpful for the diagnosis. Immunosuppressive drugs are the necessary treatment.

Effects of penehyclidine hydrochloride on Fas/FasL expression during acute lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation in rats / 中华麻醉学杂志

Objective To investigate the effects of penehyclidine hydrochloride on Fas/FasL expression during acute lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation (HSR) in rats.Methods Thirty male SPF Sprague-Dawley rats, aged 8 weeks, weighing 245-275 g, were randomly assigned into 3 equal groups using a random number table: sham operation group (group Sham) , blunt chest trauma-HSR group (group THSR) and penehyclidine hydrochloric group (group PHCD).The model of acute lung injury induced by blunt chest trauma-HSR was induced by dropping a 300 g weight onto a precordium in anesthetized rats.Blood was withdrawn via the femoral artery 5 min later until mean arterial pressure was decreased to 35-45 mmHg within 15 min, and maintained at this level for 60 min, followed by resuscitation.In PHCD group, PHCD 2 mg/kg was injected intravenously at 60 min after hemorrhagic shock.At 6 h after the model was established, the rats were sacrificed, the lungs were then removed for microscopic examination of pathologic changes and for determination of Fas, FasL and caspase-8 expression, and interleukin-6 (IL-6) and IL-1β contents in lung tissues.Apoptotic index was calculated.Results Compared with group Sham, the expression of Fas, FasL and caspase-8 was significantly up-regulated, and AI and contents of IL-6 and IL-1β were increased in THSR and PHCD groups (P＜O.05).Compared with group THSR, the expression of Fas, FasL and caspase-8 was significantly down-regulated,and AI and contents of IL-6 and IL-1β were decreased in group PHCD (P＜0.05).The pathologic changes of lungs were significantly reduced in group PHCD compared with group THSR.Conclusion The mechanism by which penehyclidine hydrochloride inhibits lung cell apoptosis induced by blunt chest trauma-HSR is associated with inhibition of Fas/FasL expression in rats.

Expression Profile of Fas-Fas Ligand in Spiral Ganglion Cells During Apoptosis

OBJECTIVES: To examine the expression profile of Fas-Fas ligand (FasL) during glutamate (Glu)-induced spiral ganglion cell (SGC) apoptosis. METHODS: Cultured SGCs were treated with 10-mM, 25-mM, and 50-mM concentrations of Glu and incubated for 24 or 48 hours. The expression intensity of FasL, Fas, caspase 3, and morphology of single SGC were evaluated using immunofluorescence staining. RESULTS: In semiquantitative analysis of the Glu-treated SGC, FasL, and caspase 3 expression intensity were increased with concentration- and time-dependent manner. Fas expression intensity did not change with different concentration at 48 hours. In morphologic analysis of the Glu-treated SGC, number of apoptotic cells were increased with concentration- and time-dependent manner. CONCLUSION: FasL was expressed in apoptotic SGCs, suggesting that the Fas-FasL signaling pathway may be involved in the Glu-induced apoptosis of dissociated SGCs.

FasL-Fas system and cerebral ischemia / 国际脑血管病杂志

Fas ligand (FasL) is critical for maintaining immune privilege status of central nervous system in physiological conditions.FasL binds Fas via extrinsic pathway mediated ischemic penumbra neuronal apoptosis after the onset of cerebral ischemia.In addition,FasL is involved in the regulation of T lymphocyte subsets by inducing the expression of inflammatory cytokines and chemokines,promoting intracerebral microglia activation and peripheral neutrophil infiltration,and thus aggravating immune inflammatory response after stroke.FasL-Fas system has a certain role for promoting the recovery of neural function during the convalescence of ischemic stroke.Taking FasL-Fas system as a target,it may become a new way for brain protection treatment of ischemic stroke.

Role of Fas/FasL signaling pathway in ulinastatin postconditioning-induced attenuation of apoptosis in myocardial cells of patients undergoing cardiac valve replacement with cardiopulmonary bypass / 中华麻醉学杂志

Objective To evaluate the role of Fas/FasL signaling pathway in ulinastatin postconditioning-induced attenuation of apoptosis in the myocardial cells of patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Forty patients of both sexes,aged 21-59 yr,of ASA physical status Ⅱ or Ⅲ (NYHA class Ⅱ or Ⅲ),scheduled for elective cardiac valve replacement with CPB,were randomly divided into 2 groups (n =20 each):control group (group C),and ulinastatin postconditioning group (group U).In group U,ulinastatin 10 000 U/kg was perfused via the aortic root at 4 000-5 000 U·kg-1 ·min-1 starting from 5 min before aortic unclamping.In group C,the equal volume of normal saline was infused instead of ulinastatin.Myocardial specimens were taken from the right auricle at 45 min after aortic unclamping for determination of Fas,Fas ligand (FasL),caspase-8,Bcl-2 and Bax expression and cell apoptosis.The ratio of Bcl-2 expression to/Bax expression (Bcl-2/Bax) and apoptotic index were calculated.Results Fas,FasL,caspase-8 and Bax expression and apoptotic index were significantly lower,and Bcl-2 expression and Bcl-2/Bax were higher in group U than in group C.Conclusion Ulinastatin postconditioning attenuates apoptosis in the myocardial cells through inhibiting Fas/FasL signaling pathway in the patients undergoing cardiac valve replacement with CPB.

Expression of FAS ligand in the ipsilateral and contralateral testicles of rats subjected to the torsion of the unilateral testicular cord

PURPOSE: To investigate the expression of FAS ligand (FASL) in ipsilateral and contralateral testicles of rats submitted to ischemia/reperfusion. METHODS: Wistar rats (n=21) distributed into groups control (GC), n=5, testicular exposure; ischemia (GI), (n=8), Torsion in the left testicular Cord (TCT) for three hours followed by orchiectomy without distortion and orchietomy of the contralateral testicle after 24 hours; and reperfusion (GR), (n=8), left TCT for 3 hours and distortion and repositioning on the scrotum and bilateral orchiectomy after 24 hours. Quantification of the FASL expression by immune-histochemistry. RESULTS: Statistical analysis showed similarity between GC and GI (p>0.05), differences detected are concentrated on the GR (p<0.05), increase in immunoexpression of FASL in the subgroups Right GR (406.8+-61.5) and Left GR (135.3 +-28.9) with significant predominance in the GR subgroup. CONCLUSION: Ischemia/reperfusion increased the FASL expression significantly in contralateral testicles in GR, in rats.

Effects of Monosialoganglioside on the Expression of Fas in Hippocampus of Cerebral Ischemia-Reperfusion Rats / 天津医药

0bjective To study the expression of Fas in hippocampus of cerebral ischemia-reperfusion rats after treatment with monosialoganglioside (GM1). Methods Seventy-two SD rats were randomly divided into sham group (n=8), GM1 treatment group (n=32) and ischemia-reperfusion (I/R) group (n=32). According to the different time points (6 h, 12 h, 24 h and 3 d) of I/R, there were four subgroups in GM1 and I/R groups respectively. The expression of Fas in hippocampus was examined by immunochemistry and Western blot methods in all groups. Results Results of immunochemistry method showed that there was a little expression of Fas (the mean optical density was 0.17±0.02) in hippocampus of sham group, and the positive expression of Fas at different time points were increased significantly after reperfusion. The mean values of opti-cal density at different time points were 0.42±0.11, 0.51±0.13, 0.55±0.13 and 0.62±0.15 in I/R group, which reached to the peak at 3 d . The positive expression of Fas at different time points were significantly decreased in GM1 group (0.29 ± 0.09, 0.34±0.11, 0.37±0.12 and 0.43±0.12) than that of I/R group (P＜0.05). Conclusion Monosialoganglioside participated in the pathogenesis of cerebral ischemia-reperfusion injury by down-regulating the expression of Fas.

Treatment of toxic epidermal necrolysis with intravenous immunoglobulin: a series of three cases / Tratamento da necrólise epidérmica tóxica com imunoglobulina endovenosa: uma série de três casos

Stevens-Johnson's syndrome (SJS) and toxic epidermal necrolysis (TEN) are life-threatening dermatoses, that lead to keratinocyte apoptosis induced by interactions between Fas (cell death receptor) and soluble Fas-ligand, present in serum of Stevens-Johnson's syndrome / toxic epidermal necrolysis patients. Anti-Fas antibodies in intravenous immunoglobulin (IVIG) would block the apoptosis cascade. Three cases of toxic epidermal necrolysis occurred in one male and two female patients, after use of allopurinol, leprosy multidrug therapy concomitant with dipyrone, and diclofenac. The cases were treated with intravenous immunoglobulin 2-3 mg/kg and prednisone 20-50 mg/day. The interruption of new lesions outbreak and reepithelization were extremely fast after the use of intravenous immunoglobulin, without adverse effects. Controlled studies are needed to confirm the efficacy of intravenous immunoglobulin in Stevens-Johnson's syndrome / toxic epidermal necrolysis, but the results seem promising.

A Síndrome de Stevens-Johnson e a Necrólise Epidérmica Tóxica são dermatoses graves, que levam à apoptose dos queratinócitos induzida pela interação entre Fas (receptor de morte celular) e Fasligante solúvel, presente no soro de pacientes com Síndrome de Stevens-Johnson e Necrólise Epidérmica Tóxica. Anticorpos anti-Fas contidos na imunoglobulina endovenosa podem bloquear esta cascata apoptótica. Três casos de Necrólise Epidérmica Tóxica são descritos, ocorrendo após uso de alopurinol, diclofenaco e poliquimioterapia para hanseníase concomitante com dipirona. Os três casos foram tratados com imunoglobulina endovenosa 2-3 mg/kg, divididos em 4 ou 5 dias e prednisona 20-50 mg/dia. A interrupção no surgimento de novas lesões e a repitelização foram extremamente rápidas, sem ocorrência de efeitos adversos. Estudos controlados são necessários para confirmar a eficácia da imunoglobulina endovenosa na Síndrome de Stevens-Johnson e Necrólise Epidérmica Tóxica, porém, seus resultados parecem ser promissores.

Quantification of Fas protein in CSF of patients with neurocysticercosis / Quantificação da proteína Fas no LCR de pacientes com neurocisticercose

Neurocysticercosis is a parasitic disease that affects the central nervous system. The objective of this study was to investigate the correlation between neuronal death evaluated by the quantification of Fas apoptotic factor and the different evolutive forms of neurocysticercosis accompanied or not by epileptic seizures. METHODS: Cerebrospinal fluid samples from 36 patients with a diagnosis of neurocysticercosis divided into the following groups: active cystic form (n=15), 9 patients with and 6 without seizures, and calcified form (=21), 9 with and 12 without seizures. Fourteen patients comprised the control group. Fas protein concentrations were determined by ELISA. RESULTS: Only the group of patients with calcified cysts without seizures presented cerebrospinal fluid levels of Fas similar to those of the control group. Higher levels were observed for the other groups. CONCLUSIONS: The present finding suggests high cerebrospinal fluid levels of soluble Fas protein, except for patients with calcified cysts without seizures. Significant differences were observed for the group with calcified cysts and seizures, suggesting greater neuronal damage in these patients. Replacement of the term inactive cyst with reactive inactive cyst is suggested.

Neurocisticercose é uma doença parasitária que afeta o sistema nervoso central. O objetivo deste estudo foi investigar a correlação entre morte neuronal por meio da quantificação do fator apoptótico Fas e a presença de neurocisticercose nas suas diferentes fases evolutivas, acompanhadas ou não de crises epilépticas. MÉTODOS: Foram analisadas amostras de líquido cefalorraquidiano em 36 pacientes com diagnóstico de neurocisticercose, determinando-se as concentrações da proteína Fas pelo método ELISA. Foram considerados os seguintes grupos: forma cística ativa n=15 (9 com crises, 6 sem crises), forma calcificada n=21 (9 com crises, 12 sem crises) e 14 pacientes (grupo controle). RESULTADOS: Apenas o grupo com calcificações sem crises apresentou níveis de Fas semelhantes ao controle. Maiores níveis foram observados nos outros grupos. CONCLUSÕES: As formas ativa e calcificada apresentam níveis elevados da proteína Fas, exceto para as formas calcificadas sem crises. No grupo de calcificações com crise, observamos diferenças mais expressivas, sugerindo maior dano neuronal. Sugerimos a substituição da denominação "cisto inativo" por "cisto inativo reagente".

The effects of zinc on the expression of liver Fas/FasL in arsenic poisoning mice / 中国地方病学杂志

Objective To investigate the effects of arsenic on liver Fas/FasL expression in mice and to observe the antagonize effect of zinc. Methods Sixty health Kunming mice were divided into five groups according to their body mass: negative control group(no arsenic and no zinc), arsenic group(55 mg/L NaAsO2 solution), low dose zinc intervention group(55 mg/L NaAsO2 solution and 20 mg/L ZnSO4 solution), middle dose zinc intervention group (55 mg/L NaAsO2 solution and 40 mg/L ZnSO4 solution), and high dose zinc intervention group (55 mg/LNaAsO2 solution and 80 mg/L ZnSO4 solution). Everyday the solution was given by oral gavage at a dose of 0.02 ml/g body weight for six weeks. The expression of Fas/FasL in mice liver was examined by immunohistochemistry.Results With the amount of zinc increasing, the expression of both Fas and FasL in mice liver decreased gradually. The expression rates of Fas[83.33%(10/12), 50%(6/12)] and FasL[66.67%(8/12), 41.67%(5/12)]in low dose zinc intervention group and middle dose zinc intervention group, respectively, were different from the expression rate of Fas[8.33%(1/12)] and FasL[0.00%(0/12)] in the negative control group(all P < 0.05). The Fas expression rate of middle dose zinc intervention group[50%(6/12)] and the high dose zinc intervention group [25%(3/12)] was compared with the arsenic group[8.33%(1/12)], and the difference was statistically significant (all P < 0.01 ). The FasL expression rate of the middle dose zinc intervention group [41.67% (5/12 )] and the high dose zinc intervention group[16.67%(2/12)] was compared with positive control group[91.67%(11/12)], and the difference was statistically significant (all P < 0.05). Conclusions Arsenic can increase the expression of Fas and FasL in mice liver, and promote apoptosis in liver, zinc may antagonize the effect of arsenic.

Inhibition of allogeneic T-cell responses by Kupffer cells expressing indoleamine 2,3-dioxygenase in vitro / 中华肝胆外科杂志

Objective To investigate kupffer cells (KCs) expressing indoleamine 2,3-dioxygenase(IDO)in the inhibition of allogeneic T-cell proliferation in vitro. Methods Real-time PCR was used to investigate the expression of IDO mRNA and FasL mRNA in KCs pretreated with or without IFNγ. High performance liquid chromatography was used to analyze the catabolism of tryptophan by IDO from KCs. Allogeneic T-cell response was used to confirm the inhibition of KCs in vitro. The proliferation of lymphocytes was detected using [3 H] thymidine incorporation. Cell cycle and lymphocyte apoptosis were evaluated by flow cytometric assay. Results Real-time PCR revealed IDO mRNA and FasL mRNA expression in KCs pretreated with IFN-γ. IDO catabolic effect was confirmed by a decrease in tryptophan and increase in kynurenine concentration. KCs expressing IDO and FasL from BABL/c mice acquire the ability to suppress the proliferation of T-cells from C57BL/6, which could be blocked by the addition of 1-methyl-tryptophan and anti-FasL antibody. The co-cultured T-cells with KCs expressing IDO and FasL could induce allogeneic T-cell apoptosis and exhibited cell-cycle arrest in G1. Conclusion In addition to the Fas/FasL pathway, IDO may also play an important role in KCs to inhibit allogeneic T-cell proliferation in vitro.

The role of Fas ligand protein in the oxidative stress induced by azoxymethane on crypt colon of rats / O papel da proteína ligante Fas no estresse oxidativo induzido pelo azoximetano em criptas do colo de ratos

PURPOSE: To study the protein Fas ligand (FasL) on the expression of apoptosis, using a model of oxidative stress induced by azoxymethane (AOM), in the crypt of colon in rats. METHODS: Wistar rats (n=14) were assigned into two groups: control (n=7) and AOM (n=7). A single subcutaneous administration of AOM (5mg/kg) or saline solution was performed at the beginning of third week and after three hours samples of proximal colon were collected. The expression of FasL was quantified (Software ImageLab) in percentage of areas in the top, base and all crypt. Results were expressed as mean ± sd (Shapiro-Wilks test and t Student test) (p < 0.05). RESULTS: In the animals of CG there was no significant difference between the FasL expression of the top (10.75±3.33) and basal (11.14±3.53) colon crypt (p=0.34293740). In the animals of AOM there was no significant difference between the FasL expression of the top (8.86±4.19) and basal (8.99±4.08) colon crypt (p=0.78486003). In the animals of CG (10.95±3.43) and AOM (8.92±4.13) there was a significant difference of the FasL expression (p=0.026466821). A significantly decrease on the FasL expression was observed in the animals of CG (10.75±3.33) and AOM (8.86±4.19) in the top crypt (p=0.00003755*). A significant decrease was also observed in the animals of CG (11.14±3.53) and AOM (8.99±4.08) in the basal colon crypt (p=0.00000381**). CONCLUSION: Azoxymethane induce the oxidative stress and the significantly decrease of FasL expression, although there is no significant difference between basal and top of colon crypt linked to consumption-activation of Fas ligand.

OBJETIVO: Avaliar o marcador de apoptose Fas ligante (FasL) em um modelo de estresse oxidativo induzido pelo azoximetano (AOM) na cripta de cólon em ratos. MÉTODOS: 14 ratos Wistar foram distribuídos em dois grupos: controle (n=7) e AOM (n=7). A AOM (5mg/kg) ou solução salina foi aplicada via subcutânea e a coleta de amostras de colo proximal ocorreu 3 horas após. A FasL foi quantificada pelo percentual de áreas no topo, base e toda a cripta. Os resultados foram submetidos aos testes de Shapiro-Wilks e t de Student (p < 0,05). RESULTADOS: No grupo GC, não houve diferença significativa entre a expressão da FasL no topo (10,75 ± 3,33) e base (11,14 ± 3,53) da cripta (p=0,34293740). No grupo AOM não houve diferença significativa entre a expressão da FasL no topo (8,86 ± 4,19) e base (8,99 ± 4,08) e cripta (p=0,78486003). No grupo GC (10,95 ± 3,43) e AOM (8,92 ± 4,13), houve uma diferença significativa da expressão da FasL (p=0,026466821). Redução significativa na expressão da FasL ocorreu nos em GC (10,75 ± 3,33) e AOM (8,86 ± 4,19) no topo da cripta (p = 0,00003755*). Foi observada diminuição significativa em GC (11,14 ± 3,53) e AOM (8,99 ± 4,08) na base da cripta (p=0,00000381**). CONCLUSÃO: Azoximetano induziu o estresse oxidativo identificado pela diminuição significativa da expressão da FasL, embora não haja diferença significativa entre a base e parte superior da cripta associada à ativação de consumo do FasL.

Involvement of the Fas and Fas ligand in testicular germ cell apoptosis by zearalenone in rat

Zearalenone (ZEA), a nonsteroidal estrogenic mycotoxin, is known to cause testicular toxicity in animals. In the present study, the effects of ZEA on spermatogenesis and possible mechanisms involved in germ cell injury were examined in rats. Ten-week-old Sprague-Dawley rats were treated with 5 mg/kg i.p. of ZEA and euthanized 3, 6, 12, 24 or 48 h after treatment. Histopathologically, spermatogonia and spermatocytes were found to be affected selectively. They were TUNEL-positive and found to be primarily in spermatogenic stages I-VI tubules from 6 h after dosing, increasing gradually until 12 h and then gradually decreasing. Western blot analysis revealed an increase in Fas and Fas ligand (Fas-L) protein levels in the ZEA-treated rats. However, the estrogen receptor (ER)alpha expression was not changed during the study. Collectively, our data suggest that acute exposure of ZEA induces apoptosis in germ cells of male rats and that this toxicity of ZEA is partially mediated through modulation of Fas and Fas-L systems, though ERalpha may not play a significant role.

The Expression of Cytochrome c, TNF-R1 and Fas Ligand in Patient's Serum after Successful Cardiopulmonary Resuscitation

PURPOSE: Numerous reports have suggested that apoptosis may play an important role in postresuscitation syndrome. The aim of this study is to assess the levels of molecules that are associated with apoptosis in the serum of patients who underwent successful resuscitation after cardiac arrest. METHODS: The serum levels of cytochrome c, tumor necrosis factor type 1 receptor (TNF-R1) and Fas ligand in 11 patients were measured at 0, 4, 12, 24, 48 and 72 hours after successful resuscitation. The primary endpoint consisted of survival to hospital discharge. Ten healthy volunteers were also evaluated as a control group. RESULTS: Patients with successful resuscitation had increased levels of cytochrome c and TNF-R1 at 0, 4, 12, 24 and 48 hours after return of spontaneous circulation (ROSC), as compared with those levels of the healthy volunteers (p<0.05). Higher levels of TNF-R1 at 12, 24 and 48 hours after ROSC were found in the non-survivors as compared to those levels of the survivors (p=0.01, 0.03, 0.02). The Fas ligand level at ROSC was also higher in the patients with successful resuscitation (p=0.00). However, the Fas ligand levels at 24, 48 and 72 hours after ROSC were lower in the patients with successful resuscitation than those levels in the healthy volunteers. CONCLUSION: These results suggest that apoptosis belonging to the TNF-R1 and cytochrome c pathways may be involved in the pathogenesis of postresuscitation syndrome. The serum levels of the death-receptor TNF-R1 could serve to quantitate the severity of injury and to prognosticate the survival outcomes.