ObjectiveTo investigate the clinical values of serum histidine decarboxylase (HDC), D-lactate, and alpha-glutathione S-transferase (α-GST) for diagnosing intestinal mucosal injury of patients with intestinal obstruction. MethodsThe expression levels of serum HDC, D-lactate, and α-GST in 28 patients with strangulated intestinal obstruction, 19 patients with simple intestinal obstruction, 17 patients with acute simple appendicitis, and 20 healthy volunteers were determined by enzyme linked immunosorbent assay (ELISA) before the treatment, and then the area under receiver operating characteristic curve (AUC) of these diagnostic indices were compared. In addition, the occurrence rates of systemic inflammatory response syndrome (SIRS) and infectious complications (abdominal cavity infection and pulmonary infection) were closely observed. The relevances of SIRS and infectious complications and the expression levels of these three diagnostic indices were analyzed. ResultsThe expression levels of serum HDC, D-lactate, and α-GST of the patients with strangulated intestinal obstruction were the highest among all the patients (Plt;0.01), and the expression levels of these three indices in the patients with simple intestinal obstruction were higher than those of the patients with acute simple appendicitis (Plt;0.05). The AUC of HDC (0.913) was larger than that of D-lactate (0.872) and α-GST (0.836) (P=0.000, P=0.000, respectively). When the cut off value of HDC was 31.00 μg/L, the sensitivity, specificity, false negative rate, and false positive rate of HDC were 74.5%, 94.6%, 25.5%, and 5.4%, respectively, which were all better than those of D-lactate and αGST. The occurrence rates of SIRS and abdominal cavity infection of the patients with strangulated intestinal obstruction were significantly higher than those of patients with simple intestinal obstruction (P=0.046) and acute simple appendicitis (P=0.027); while there was not significantly different of pulmonary infection among all the patients (P=0.728). The expression level of serum HDC in patients with strangulated intestinal obstruction suffered from SIRS (P=0.000) or abdominal cavity infection (P=0.002) was significantly higher than that of not-suffered from SIRS or uninfected patients. Meanwhile, the expression levels of serum D-lactate and α-GST in the patients with strangulated intestinal obstruction suffered from SIRS were higher than those of notsuffered from SIRS patients (P=0.032, P=0.021, respectively). The expression levels of HDC, D-lactate, and α-GST were significantly correlated with SIRS and abdominal cavity infection (Plt;0.05), among which the level of HDC and the incidence of SIRS had the highest correlation (r=0.608, P=0.001). ConclusionHDC may be a more effective index for diagnosing intestinal mucosal injury of patients with intestinal obstruction.
Objective To investigate the relationship between the expressions of P-gp, GST-π and C-erbB-2 and clinicopathologic characteristics as well as prognosis in breast cancer. Methods The expressions of P-gp， GST-π and C-erbB-2 were detected by immunohistochemistry in 48 cases of breast cancer, and histopathologic characteristics as well as 5-year survival rate of these cases were analyzed. Results There was no significant difference in the expressions of P-gp and GST-π with age, histologic grade, number of lymph node metastasis and TNM stage of breast cancer ( P ＞ 0.05). There was significant difference in expression of C-erbB-2 with histologic grade, number of lymph node metastasis and TNM stage of breast cancer （ P ＜ 0.05）. Positive rate of P-gp expression in breast cancer with positive C-erbB-2 expression was remarkably higher than that in breast cancer with negative C-erbB-2 expression ( P ＜ 0.05) . Positive rate of GST-π and C-erbB-2 expression in survivals within 5 years was remarkably lower than that in deaths within 5 years ( P ＜ 0.01). Conclusion P-gp participates primary drug-resistance mechanism of breast cancer. The possibility of primary drug-resistance is higher in breast cancer with positive C-erbB-2 expression. The expression of C-erbB-2 helps to evaluate prognosis and the result of treatment in breast cancer.
Objective To explore changes of 3’-glutamylcysteine synthetase( γ-GCS)and reduced glutathione(GSH)in patients with bronchial asthma．Methods Ten patients with acute asthma were enrolled and treated for six weeks according to guideline recommendations．Levels of -GCS，GSH and malondialdehyde(MDA)in total cells in induced sputum and GSH，MDA，reactive oxygen(ROS)in selum were measured and compared before and after therapy．Ten healthy volunteers were as normal contro1．Meanwhile，the pulmonary function(FEVl％pred)was measured and asthmatic symptoms were quantified using Hogg’s way．Results A．In serum and sputum of the asthma patients，GSH were lower and MDA were higher before treatment than those of the control(Plt;0．01)．And -GCS in induced sputumwere higher before treatment than those of contro1．B．After treated for six weeks．levels of GSH in serum and sputum of the asthma patients increased copmpared to baseline(all Plt;0．01)，but were still lower than that of control(Plt;0．05)．Activities of MDA in serum and sputum and -GCS in sputum were elevated compared to baseline(Plt;0．01)，but still higher than that of control(all Plt;0．05)．C．Levels of GSH in serum of all patients were correlated negatively witll asthmatic symptom scores and levels of MDA and ROS(r=－0.701，－0.901，－0.878；Plt;0.05，lt;0.01，lt;0.01)．There was a positive relationship between levels ofGSH in serum and FEV1％pred(r=0．854，Plt;0．01)．In induced sputum，activities of 3’-GCS in all patients was correlated positively with their asthmatic symptom scores and level of MDA f r=0．804，0.926；Plt;0.05，lt;0.叭)．Conclusion γ-GCS and GSH may participate the reaction of
The aim of this research is to investigate the influence of microencapsulation on the expression of the oxidative stress genes and exogenous regulation of HepG2 cells. We compared the expression of hemeoxygenase-1 (HO-1) and glutathione S-transferases-A1 (GST-A1) in HepG2 cells under different culture conditions through real-time PCR. The effects of exogenous antioxidants on cell viability and albumin levels were also evaluated through MTT assay and ELISA assay. The results showed that after culturing for 6 and 16 days, the expression levels of HO-1 in encapsulated cells were approximately 4.9 and 3.1 times higher than that of monolayer cells at the same culture period; As for the expression levels of GST-A1, they were elevated to 11.2 and 33 times of monolayer cells (P<0.05). Accordingly, we found that NAC at 5-10 mmol/L significantly increased the viability by 40%-70% and the biosynthetic function by 20%-30% in microencapsulated HepG2 cells (P<0.05). GSH increased the viability of the encapsulated cells by 20%-55% and the biosynthetic function by 15% (P<0.05). In conclusion, oxidative stress exists in the microcapsules and affects genes expression. Exogenous antioxidants can prevent the inhibition effects of oxidative stress on cellular growth.
ObjectiveTo explore the antioxidant effects of reduced glutathione on rat pulmonary fibrosis compared with traditional corticosteroid. MethodsOne-hundred and eight healthy SD rats were randomly divided into 6 groups,ie. a control group,a model group,a dexamethasone group,a low-dose glutathione group,a middle-dose glutathione group,and a high-dose glutathione group,with 18 rats in each group. The pulmonary fibrosis model was established by intratrachially instillation of bleomycin in all rats except the control group. The severity of lung fibrosis was evaluated by HE staining and Masson staining of collagen,and measurement of glutathione,hydroxyproline,superoxide dismutase (SOD),glutathion peroxidase (GSH-Px)in lung tissue homogenate by photocolorimetric method. ResultsOn 7th day and 14th day after bleomycin instillation, the severity of alveolitis in the model group,the dexamethasone group,and three glutathione intervention groups was significantly reduced compared with the control group (P<0.05). On 28 day, the severity of lung fibrosis was significantly reduced in the dexamethasone group and three glutathione intervention groups compared with the model group (P<0.05). On 7th day,lung glutathione content was significantly lower in the model group compared with the control group (P<0.05), significantly higher in the dexamethasone group and three glutathione intervention groups compared with the model group (P<0.05), significantly lower in the dexamethasone group and the low-dose glutathione group compared with the control group (P<0.05), and significantly higher in the high-dose glutathione group compared with the dexamethasone group (P<0.05). On 7th,14th,and 28th day,the hydroxyproline content in the dexamethasone group and three glutathione intervention groups decreased significantly compared with the model group (P<0.05). On 14th day,the hydroxyproline content in the middle-dose and high-dose glutathione groups was significantly lower than that in the dexamethasone group (P<0.05). SOD and GSH-Px were significantly reduced in the model group compared with the control group on all time points (P<0.05),but significantly increased after intervention by different doses of glutathione (P<0.05). ConclusionReduced glutathione can significantly reduce the degree of pulmonary fibrosis in rats,but has no obvious advantage over dexamethasone.
Objective To investigate the protective effect of the antioxidant glutathione (GSH) on the steroid-induced imbalance between osteogenesis and adipogenesis in human bone marrow mesenchymal stem cells (BMSCs). Methods The BMSCs were isolated from the proximal femur bone marrow from 3 patients of femoral neck fracture and were separated, cultured, and purificated by density gradient centrifugation and adherent wall methodin vitro. The third generation BMSCs were divided into 5 groups: group A, BMSCs (1×105 cells/mL); group B, BMSCs (1×105 cells/mL)+10 μmol/L dexamethasone; group C, BMSCs (1×105 cells/mL)+10 μmol/L dexamethasone+5 μmol/L GSH; group D, BMSCs (1×105 cells/mL)+10 μmol/L dexamethasone+10 μmol/L GSH; group E, BMSCs (1×105 cells/mL)+10 μmol/L dexamethasone+50 μmol/L GSH. After cultured for 7 days, the reactive oxygen species expression was detected by flow cytometry; the superoxide dismutase (SOD) and Catalase mRNA expressions were determined by RT-PCR; the peroxisome proliferator-activated receptors γ (PPAR-γ), CCAAT/enhancer-binding family of proteins (C/EBP), Runx2, and alkaline phosphatase (ALP) mRNA expressions were evaluated by real-time fluorescence quantitative PCR. After cultured for 21 days, Oil red O staining was used to observe the adipogenesis differentiation of cells, and the expressions of related proteins were detected by Western blot. Results The reactive oxygen species expression in group B was obviously higher than in the other groups, in group C than in groups A, D, and E, and in groups D, E than in group A, all showing significant differences between groups (P<0.05); but there was no significant difference between groups D and E (P>0.05). The oil red O staining positive cells in group B were obviously more than the other groups, and groups C, D, E, and A decreased sequentially, the absorbance (A) values had significant differences between groups (P<0.05). RT-PCR detection showed that the relative expressions of SOD and Catalase mRNA in group B were significantly lower than those in the other groups, while in group C than in groups A, D, and E (P<0.05), but there was no significant difference among groups A, D, and E (P>0.05). Real-time fluorescence quantitative PCR detection showed that the relative expressions of PPAR-γ and C/EBP mRNA in group B were significantly higher than those in the other groups, while in group C than in groups A, D, and E, and in groups D, E than in group A (P<0.05); but there was no significant difference between groups D and E (P>0.05). The relative expressions of Runx2 and ALP mRNA in group B were significantly lower than those in the other groups, while in group C than in groups A, D, and E, and in groups D, E than in group A (P<0.05); but there was no significant difference between groups D and E (P>0.05). Western blot detection showed that the relative expression of PPAR-γ and C/EBP protein in group B was significantly higher than those in the other groups, and groups C, D, E, and A decreased sequentially, all showing significant differences between groups (P<0.05). The relative expression of Runx2 and ALP protein in group B was significantly lower than those in the other groups, and groups C, D, E, and A increased sequentially, all showing significant differences between groups (P<0.05). Conclusions GSH can inhibit the adipogenesis differentiation and enhance the osteogenic differentiation of human BMSCs by reducing the intracellular reactive oxygen species level; and in a certain range, the higher the concentration of GSH, the more obvious the effect is.