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Objective To investigate the effects of artesunate (AS) on septic lung injury in mice and to study the modulation of heme oxygenase-1 (HO-1) in lung in order to clarify the mechanism of AS action.Methods Sixty male Kunming mice were randomly (random number) divided into four groups: Sham group (n =15), CLP group (n =15), AS + CLP group (n =15) and AS + ZnPP + CLP group (n =15).Cecal ligation and puncture (CLP) method was employed to induce septic lung injury.AS (15 mg/ kg) was injected into the abdomen of mice 2 hours before the CLP procedures, and ZnPP Ⅸ, an inhibitor of HO-1, was intraperitoneally injected in dose of 40 μmol/kg 1 hour after the AS injection.The equivalent volume of normal saline was intraperitoneally injected instead in mice of Sham group and CLP group.The mice were sacrificed 24 hours after the CLP procedures.The TNF-α, IL-6 in serum were assayed by ELISA method.The lung injury score and wet/dry ratio were measured.The western blotting and immunohistochemistry methods were used to determine HO-1 protein expression in lung tissue.The protein level of nuclear factor-E2-related factor-2 (Nrf-2), an important transcriptional factor of HO-1 in lung tissue was also analyzed by western blotting.One-way analysis of variance (ANOVA) was used for comparisons among the groups, and SNK-q (Student-Newman-Keuls) test was performed for further comparison, and difference was statistically significant at P ＜ 0.05.Results The TNF-α (pg/mL) (54.37 ± 15.59 vs.627.45 ± 117.03, P ＜ 0.05), IL-6 (pg/mL) (81.53 ± 26.89 vs.898.52 ± 222.78, P ＜ 0.05) in serum was increased, and the lung protein exudation, pulmonary edema (wet/dry weight ratio: 4.27 ± 0.22 vs.6.78 ±0.73, P ＜ 0.05), pulmonary pathology injury (lung injury score: 2.20 ± 0.2 vs.13.25 ± 2.67, P ＜ 0.05) were aggravated by CLP.The HO-1 and Nrf-2 were up-regulated in lung tissue in CLP group compared with the sham group (P ＜ 0.05).After the intervention of AS, the HO-1 and Nrf-2 were further increased (P＜0.05), theTNF-α (pg/mL) (627.45 ±117.03 vs.307.88 ±72.33, P＜0.05), IL-6 (pg/mL) (898.52 ± 222.78 vs.413.47 ± 115.14, P ＜ 0.05) in serum, lung protein exudation, pulmonary edema (wet/dry weight ratio: 6.78 ± 0.73 vs.5.05 ± 0.61, P ＜ 0.05), pulmonary pathology injury (lung injury score: 13.25 ± 2.67 vs.4.95 ± 1.46, P ＜ 0.05) were attenuated compared with the CLP group.However, the protective role of AS in the septic lung injury in mice was partly reversed by ZnPP, and no significant difference was detected between the AS + CLP + ZnPP and CLP group (lung injury score: 12.15 ± 2.95 vs.13.25 ± 2.67, P ＞ 0.05;wet/dry weight ratio: 6.78 ± 0.73 vs.6.29 ± 0.82, P ＞ 0.05).Conclusions AS plays protective roles in septic lung injury, and it is attributed to limiting lung inflammation via up-regulation of HO-1.
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Objective To observe the apoptosis or oncosis of pancreatic acinar cells of different severity of acute pancreatitis (AP) and the release level of enzymes in vitro, and to investigate the relationship between them. Methods Two-step enzymatic digestion method was used to separate pancreatic acinar cells into 4 groups. 0. 1 μg/ml of the caerulein was added in the AP group. Caerulein and LPS (bacterial lipopolysaccharide, 10 mg/L) were added in LPS group. Caerulein and OCT (octreotide, 100 ng/ml) were added in OCT group. Medium was added in the control group. AO (acridine orange) and EB (ethidium bromide) double staining method was used to detect the incidence of apoptosis or oncosis of acinar cell. The release of intracellular enzyme was detected by measuring the concentrations of amylase and LDH in cell culture media by colorimetry method. Results The apoptosis index was 2.2 + 0.4, 6.4 ± 0.6, 4.6 + 0.4, 11.2 +1.2 in the control group, AP group, LPS group, OCT group; while the oncosis index was 3.0 +0.4, 17.2 ±1.6, 23.0 ± 2.2, 12.8 ± 1.4 in the control group, AP group, LPS group, OCT group; the release of LDH was (2180 ±240), (8060 ±930), (9460 +920), (6860 ±740) U/dl, the level of amylase was (1750 ± 190),(3820 ±460), (4420 ±480), (2260 ±260)U/L. All the values in the experiment groups were significantly higher than that in control group ( P ＜ 0.05 ). The oncosis index, LDH, amylase in LPS group was significantly higher than that in AP group ( P ＜ 0.05 ), but the apoptosis index in LPS group was significantly lower than that in AP group ( P ＜ 0.05 ). The apoptosis index in OCT group was significantly higher than that in AP group ( P ＜ 0. 05 ), but the oncosis index, LDH, amylase was significantly lower than that in AP group ( P ＜ 0. 05 ).Conclusions Induction of apoptosis and reduction of oncosis in AP pancreatic acinar cells can reduce the release of enzyme in acinar cells.
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Nonspecific inflammatory response is the major cause for failure of islet grafts at the early phase of intraportal islet transplantation (IPIT). Bilirubin, a natural product of heme catabolism, has displayed anti-oxidative and anti-inflammatory activities. The present study has demonstrated that bilirubin protected islet grafts by inhibiting nonspecific inflammatory response in a syngeneic rat model of IPIT. The inflammation-induced cell injury was mimicked by exposing cultured rat insulinoma INS-1 cells to cytokines (IL-1beta, TNF-alpha and IFN-gamma) in in vitro assays. At appropriate lower concentrations, bilirubin significantly attenuated the reduced cell viability and enhanced cell apoptosis induced by cytokines, and protected the insulin secretory function of INS-1 cells. Diabetic inbred male Lewis rats induced by streptozotocin underwent IPIT at different islet equivalents (IEQs) (optimal dose of 1000, and suboptimal doses of 750 or 500), and bilirubin was administered to the recipients every 12 h, starting from one day before transplantation until 5 days after transplantation. Administration of bilirubin improved glucose control and enhanced glucose tolerance in diabetic recipients, and reduced the serum levels of inflammatory mediators including IL-1beta, TNF-alpha, soluble intercellular adhesion molecule 1, monocyte chemoattractant protein-1 and NO, and inhibited the infiltration of Kupffer cells into the islet grafts, and restored insulin-producing ability of transplanted islets.
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Animals , Male , Rats , Apoptosis/drug effects , Bilirubin/administration & dosage , Cell Line, Tumor , Cytokines/immunology , Diabetes Mellitus, Experimental/drug therapy , Inflammation , Inflammation Mediators/immunology , Islets of Langerhans/drug effects , Islets of Langerhans Transplantation , Oxidative Stress/drug effects , Rats, Inbred LewABSTRACT
uld remarkably inhibit the systemic inflammatory reaction and attenuate the severity of pancreatic injury,and the mechanism may include inhibit the release of TNF-α and IL-1 as well as up-regulate the production of IL-10.
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objectlve To investigate the efficacy of continuous low-dose fluorouracil with cetuximab for antiangiogenic effect on colon carcinoma xenograft,and test its antitumor effect and toxicity.Methods Balb/c mice bearing CT-26 colon carcinoma xenograft were randomly divided into five groups,receiving low-dose metronomic(LDM)fluorouracil,maximum tolerated dose(MTD)fluorouracil,cetuximab,LDM fluorouracil with cetuximab therapy and saline respectively.Tumor growth,weight loss,peripheral white blood cell counts and survival of mice were monitoted.At the end of experiment,tumors were resected for tumor microvascular density(MVD)by immunofluorescence staining. Results Tumor growth inhibition was found in mice receiving LDM fluorouracil therapy and combined therapy,without significant body weight loss or leukopenia,and the survival of mice was remarkably prolonged,compared with mice receiving MTD fluorouracil or cetuximab therapy,and the antitumor effects of the combined therapy was stronger than that of the fluorouracil LDM therapy.LDM treatment and combine treatment led to statistically significant(P＜0.05)55%and 71%reduction in tumor growth,as well as 73%and 77% reduction in tumor microvessel density compared with the control respectively.Additonally,tunnel staining shows no significant difference between these treatment groups. Conclusion Continuous low-dose regimen of fluorouracil with cetuximab can significantly increase the therapeutic activity with decreased toxicity and prolonged animal survival bearing implanted colon cancer.
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Objective:The study aims to investigate the expression of integrin ?V?3 and its relationship in gastric cancers.Methods:The tumoral and normal tissues were collected from 65 cases of gastric cancer patients and 40 controls.The clinical phasing and pathological staging of the patients were recorded,and immunohistochemical analysis was used to detect the expression of integrin ?V?3 and microvessel density(MVD)in gastric cancers.Results:The positive rate of integrin ?V?3 expression was significantly higher in gastric cancer tissues than normal mucosal tissues(72.38% vs 22.86%,?2=8.35,P=0.032).There was a significant difference in integrin ?V?3 expression or MVD between tumors of infiltrating and expanding growth patterns(?2=14.97,P=0.002;t=10.150,P=0.001,respectively),cancers in T3～T4 and T1～T2 stages(?2=15.21,P=0.015;t=5.961,P=0.001,respectively).The positive rate of integrin ?V?3 expression and MVD also showed corelationship with lymph node invasion and metastases.Conclusion:Tumoral expression of integrin?V?3 was closely related with biological behavior of gastric cancers,suggesting that detection of integrin ?V?3 expression may present a potential tool to evaluate the clinical and pathological staging,lymph node status and metastases.
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<p><b>OBJECTIVE</b>The -455 G/A (HaeIII) polymorphism of beta-fibrinogen gene influences levels of plasma fibrinogen. We further investigated whether it influences the risk of ischemic cerebrovascular disease.</p><p><b>METHODS</b>We accumulated 134 acute ischemic cerebrovascular disease (ICVD) cases and compared their -455 G/A status with a control group (n = 166). The beta-fibrinogen gene -455 G/A polymorphism was analyzed for all subjects by PCR-RFLP with the restrictive enzyme HaeIII.</p><p><b>RESULTS</b>Plasma fibrinogen was higher in AA homozygous participants (341 mg/dL) than in participants carrying the G allele: GA (290 mg/dL), GG (298 mg/dL) in the control group. Plasma fibrinogen was also higher in AA homozygous patients (353 mg/dL) than in cases carrying the G allele: GA (287 mg/dL), GG (302 mg/dL) in the ICVD group. However, there was no significant association between beta-fibrinogen gene -455 G/A polymorphism and ICVD group.</p><p><b>CONCLUSIONS</b>Although a small effect cannot be excluded, beta-fibrinogen gene -455 G/A polymorphism is an independent predictor of plasma fibrinogen, but not of ischemic cerebrovascular disease.</p>
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Female , Humans , Male , Middle Aged , Alleles , Cerebrovascular Disorders , Blood , Genetics , Fibrinogen , Genetics , Metabolism , Genetic Predisposition to Disease , Homozygote , Polymorphism, Restriction Fragment Length , Risk FactorsABSTRACT
Objective To study the apoptosis-inducing effect of recombinant adeno-associated virus serotype 8 with soluble TRAIL gene on human colon cancer cell line HCT116. Method Recombinant adeno-associated virus serotype 8 with TRAIL gene was generated by triple plasmid transfection of HEK293 cells with phosphate calcium precipitation. After infected by recombinant virus, cell growth assay was carried out by counting alive cells after trypan blue exclusion. Alternations of cell morphology was examined by microscopy. Cell apoptosis was detected by DNA laddering. The protein expressions of apoptosis-3 and apoptosis-8 were detected by Western blotting. Result The growth rate of the AAV8-sTRAIL infected HCT116 cells was significantly inhibited. Recombinant virus induced apoptosis in colon tumor cells by triggering caspase cascade, but not in normal human hepatocytes. Conclusion AAV8-sTRAIL efficiently suppresses the growth of human colon cancer cell line HCT116 by inducing apoptosis, with no evident toxicity to normal cell.
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Objective To study the inhibitory effect of arsenic trioxide(As_2O_3) on the tumor growth of breast cancer cell line MCF-7 implanted subcutaneously in nude mice and its mechanism.Methods BALB/C-nu/nu nude mice were subcutaneously injected with MCF-7 breast cancer cell line,and treated with intraperitoneal injection of As_2O_3 and 5-FU in different concentrations.The implanted tumor was weighed,and the tumor inhibition rates were calculated.The apoptosis of the implanted tumor was detected by flow cytometry.The expressions of bcl-2 and Fas induced by As_2O_3 were examined by immunohistochemical method.Routine blood test and bone marrow test were used to observe the function of hematopoietic system after As_2O_3 treatment.Results The growth of implanted tumor was markedly inhibited with 5-FU,low dose and high dose As_2O_3,the inhibitory rates being 38.33%、51.42% and 62.43%,respectively.The inhibitory effect of As_2O_3 was significantly stronger than that of 5-FU(P
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Objective To explore the clinical significance of micro-metastasis ( mM ) in the nipple-areola complex (NAC) and the regional skin of breast cancer. Methods Samples from the skin projection of the lump and the midline-transection of the nipple-areola complex were collected from 60 breast cancer patients for both routine pathological examination ( RP) and cytokeratin-19 (CK-19) monoclonal antibody immuneohistochemical examination (IHC). Results NAC invasion was identified by RP in 3 cases (5. 0% ) , and by IHC in 7 cases (11.7%) ( x2 = 2. 25, P