Glyoxal damages human aortic endothelial cells by perturbing the glutathione, mitochondrial membrane potential, and mitogen-activated protein kinase pathways.

BACKGROUND: Exposure to glyoxal, the smallest dialdehyde, is associated with several diseases; humans are routinely exposed to glyoxal because of its ubiquitous presence in foods and the environment. The aim of this study was to examine the damage caused by glyoxal in human aortic endothelial cells. METHODS: Cell survival assays and quantitative fluorescence assays were performed to measure DNA damage; oxidative stress was detected by colorimetric assays and quantitative fluorescence, and the mitogen-activated protein kinase pathways were assessed using western blotting. RESULTS: Exposure to glyoxal was found to be linked to abnormal glutathione activity, the collapse of mitochondrial membrane potential, and the activation of mitogen-activated protein kinase pathways. However, DNA damage and thioredoxin oxidation were not induced by dialdehydes. CONCLUSIONS: Intracellular glutathione, members of the mitogen-activated protein kinase pathways, and the mitochondrial membrane potential are all critical targets of glyoxal. These findings provide novel insights into the molecular mechanisms perturbed by glyoxal, and may facilitate the development of new therapeutics and diagnostic markers for cardiovascular diseases.

An infectious molecular clone in early infection with HIV-1 subtype CRF01_AE strains: construction and biological properties.

Our aim was to construct infectious molecular clones of the CRF01_AE subtype in the primary infection phase of an acute HIV-1 infections in people screened from MSM populations, as well as continue preliminary research on this virus and its biological properties pertaining to deriving viruses. Walking sequencing was performed on a half-molecular clone with target fragment inserted. Western Blot was used to detect protein expression in HIV-1 infected 293T cells. Sequence analysis of HIV-1 genomic clones showed full-length HIV-1 genomic clones without frame shift mutation or termination codon. HIV-1 p24 antigens generated from 08-IMC were slightly greater than those from infectious molecular clones pNL4-3 3 and 93JP-NH1, but without statistical difference (all P > 0.05). The relative light units of 08-ISO was higher than those of 08-IMC, but no significant difference was observed (all P > 0.05). 08-IMC-driven virus was linked to lower replication kinetics. The replication levels of pNL4-3 and 08-ISO were significantly higher than the 08-IMC replication level but close to NH1 replication level (all P < 0.05). 08-IMC could infect the cells expressing CCR5 and be replicated in the CCR5-expressing cells with a positive percentage of 24.3 %, 08-ISO may use CCR5-using macrophage-tropic isolates as coreceptor, while pNL4-3 viruses with T cell tropisms utilize the CXCR4 co-receptor. Our study showed that the infectious molecular clones of viruses in the primary infection phase have a close relationship with the major prevalent CRF01_AE strains and have high homology with the viral RNA in plasma.

[Urinary neutrophil gelatinase-associated lipocalin (uNGAL) and early diagnosis of paraquat poisoning patients with acute kidney injury].

OBJECTIVE: To explore the use of the urinary neutrophil gelatinase associated lipocalin (uNGAL) in the early diagnosis of paraquat poisoning patients with acute kidney injury (AKI). METHODS: Eighty five patients were from the emergency department in our hospital. Five ml blood and urine were collected from each patient at 15 min, 2, 4, 6, 8, 10, 12, 18, 24, 36, 48 and 72 h, 5 and 7d after admission. The uNGAL levels of urine were detected with ELISA test and the SCr levels were measured with creatine oxidase assay. RESULTS: Sixty two cases of paraquat intoxication suffered from AKI, the incidence was 72.94% (62/85). The SCr levels of 62 cases with AKI at 18, 24, 36, 48, 72 h and 5, 7 d after admission increased significantly, as compared with the baseline value and control group (P < 0.01). At 24, 36, 48, 72 h and 5, 7 d after admission, there was significant difference of the SCr levels between AKI group and non-AKI group (P < 0.01). At 2 h after admission, the uNGAL level of urine in paraquat intoxication AKI group was (96.21 +/- 45.32) microg/L which was significantly higher than the baseline value. At 10, 12, 18, 24, 36, 48, 72 h and 5, 7 d after admission, the uNGAL levels of urine in AKI group and non-AKI group obviously enhanced, as compared with the baseline value and control group (P < 0.01 or P < 0.05). At all time points, there was significant difference of the uNGAL level between AKI group and non-AKI group (P < 0.01). CONCLUSION: The uNGAL level of urine in paraquat intoxication patients at 2 h after admission significantly enhanced, which is earlier than enhanced SCr. So the uNGAL level of urine may serve as early diagnostic biomarker for AKI induced by paraquat intoxication.

[Clinical study of dynamic change of inflammatory factors in serum of acute paraquat rats].

OBJECTIVE: To investigate the change of inflammatory factor in lung tissue of acute paraquat (PQ) poisoned rats. METHODS: hundred SD rats were randomly divided into two groups: the normal control group (NC group, n = 10) and the PQ group (n = 80). The 1 ml saline was administered once in normal control group. The PQ group was administered with 25 mg/kg 1% PQ by intraperitoneal injection to establish the model of PQ induced renal injury. At six hours, at the first, the third and the seventh day the PQ group were sacrificed, while at the first day the normal control group was sacrificed. The level of normal tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), IL-6 in serum of rats were detected. Meanwhile, pathological changes of the renal were examined under optical microscope. RESULTS: Histopathological findings of an earlier, a large number of patients edema clearly inflammatory cell infiltration. Compared with the control group, PQ exposure of serum TNF-α, IL-2, IL-6, the level at each time point were elevated. PQ treated group 6 h and 1, 3, 7 d when the IL-2 levels were (2.16 ± 0.65), (2.95 ± 1.02), (3.05 ± 1.12), (2.21 ± 0.62) µg/L, IL-6 were (62.5 ± 8.6), (85.6 ± 13.5), (90.3 ± 15.6), (65.3 ± 9.1) ng/ml, TNF-α were (1.95 ± 0.53), (2.86 ± 0.92), (3.15 ± 1.02), (2.06 ± 0.71) µg/L, compared with the control group, are significantly higher, the differences were statistically significant (P < 0.01). CONCLUSION: acute PQ poisoning serum TNF-α, IL-2, IL-6 levels were significantly increased both early and late inflammatory factors involved in PQ poisoning the pathogenesis of renal injury.

miRNA-221 promotes proliferation, migration and invasion by targeting TIMP2 in renal cell carcinoma.

INTRODUCTION: MicroRNAs (miRNAs) play important roles in tumorigenesis. In this study, we investigated the role of miR-221 in the development and progression of clear cell renal cell carcinoma (ccRCC). METHODS: Quantitative real-time PCR (qRT-PCR) was used to measure the expression level of miR-221 in ccRCC tissues and cell lines. Then, we investigated the role of miR-221 to determine its potential roles on renal cancer cell proliferation, migration and invasion in vitro. A luciferase reporter assay was conducted to confirm the target gene of miR-221 and the results were validated in renal cancer cells. RESULTS: In the present study, we found that miR-221 was significantly increased in ccRCC tissues and cell lines. Knocked-down expression of miR-221 remarkably inhibited cell proliferation, migration and invasion of renal cancer cells. Moreover, at the molecular level, our results suggested that TIMP2 as a direct target of miR-221 through which miR-221 promoted tumor cell proliferation, migration and invasion. CONCLUSIONS: These findings suggested that miR-221 play an oncogenic role in the renal cancer cell proliferation, migration and invasion by directly inhibiting the tumor suppressor TIMP2, indicating miR-221 act as a potential new therapeutic target for the treatment of ccRCC.

[Clinical study on role of CD34 in patients with Biphenotypic acute leukemia].

This study was aimed to explore the expression of CD34 in patients with biphenotypic acute leukemia (BAL) and its relation with the prognosis of BAL. The flow cytometry was used to detect leukemia-associated antigen. The used monoclonal antibodys (McAb) included CD10, CD19 and CD34 for B lymphocyte lineage, CD2, CD3 and CD5 for T lymohocyte lineage, MPO, CD13 and CD33 for myeloid lineage. The finally results were respectively analyzed. The results indicated that 9 out of 216 cases of leukemia was diagnosed as BAL (4.2%). Among 9 cases of BAL, 6 cases showed the common expression of myeloid and T lymohocyte lineages (66.7%), 3 cases showed the common expression of myeloid and B lymohocyte lineages (33.3%). 4 cases of BAL displayed CD34 positive expression (44.4%). As compared with acute myelogenous leukemia (AML) and acute lymphoblastic leukemia (ALL), the BAL patients showed higher CD34 positive expression (P < 0.05). It is concluded that the BAL patients show a poor prognosis, as compared with AML or ALL patients. The therapeutic effect of BAL may negatively correlate with the CD34 positive expression.

Down-regulation of long non-coding RNA LET is associated with poor prognosis in gastric cancer.

INTRODUCTION: Long non-coding RNAs (lncRNAs) have emerged recently as major players in tumor biology and may be used for cancer diagnosis, prognosis, and potential therapeutic targets. Although down-regulation of lncRNA LET in several cancers has been studied, its role in gastric cancer remains unknown. The aim of our study was to investigate the expression, and clinical significance of lncRNA LET in gastric cancer. METHODS: The expression of lncRNA LET was detected by quantitative real-time PCR (qRT-PCR) in pairs of tumor tissues and adjacent non-tumor tissues of 93 gastric cancer patients. Then, we analyzed the potential relationship between lncRNA LET expression levels in tumor tissues and clinicopathological features of gastric cancer, and clinical outcome. RESULTS: We found that lncRNA LET expression was markedly down-regulated in tumor tissues compared with adjacent non-tumor tissues, and associated with depth of invasion, lymph node metastasis, distant metastasis, and TNM stage. Kaplan-Meier analysis showed that patients with low lncRNA LET expression had a poor overall survival than those with high lncRNA LET expression. Moreover, univariate and multivariate analyses showed that low lncRNA LET expression was an independent poor prognostic factor for gastric cancer patients. CONCLUSIONS: Our data provided the first evidence that lncRNA LET might be a novel prognostic indicator in gastric cancer and might be a potential target for diagnosis and gene therapy.