[Changes of the expression for genes related with senescence and the telomerase activity during cellular replicative and premature senescence in human embryonic lung fibroblasts].

Objective: To detect the alterations of telomerase activity and the expression for oxidative stress responsive genes related with senescence during cellular replicative senescence and hydrogen peroxide-induced premature senescence in human embryonic lung fibroblasts (HELFs) in vitro. Methods: The HELFs were divided into young cells (22 population doubling levels, 22PDL) , mid-aged cells (35PDL) and replicative senes-cent cells (49PDL) and premature senescent cells induced by H(2)O(2)(premature senescence, PS). The telomerase activity was detected by ELISA assay during cellular replicative and premature senescence. The mRNA level of oxidative stress responsive genes related with senescence for Foxo1, Foxo3, Pdx1, apoA-I and MMP1 was per-formed by RT-Q-PCR separately. Results: The mRNA level for Foxo1, Foxo3, apoA-I and Pdx1 was decreased separately during cellular replicative senescence compared to that in the young-stage cells with statistical signifi-cance (P<0.05). The expression of MMP1 was up-regulated 5.1-fold obviously (P<0.05). In premature senes-cence, the mRNA level was only decreased for Foxo1, Foxo3 and apoA-I, but up-regulated 2.3-fold and 6.2-fold for Pdx1 and MMP1 respcetively vs 22PDL significantly (P<0.05). The telomerase activity in young cells was not detected, and it increased in mid-aged cells and replicative senescence stages during cellular replicative se-nescence as compared to 22PDL with statistical significance (P<0.05). The telomerase activity in premature se-nescence was highly active. Conclusion: The expression for genes related with senescence has differences be-tween replicative and premature senescence and hydrogen peroxide modifies their expression levels. The telomer-ase activity has been going up with increased PDLs.

Effect of Monochromic Light-emitting Diode Light with Different Color on the Growth and Reproductive Performances of Breeder Geese.

The purpose of this study was to investigate the effect of monochromic light-emitting diode (LED) light with different color on the growth and reproductive performances of white Roman breeder geese. A randomized complete batch design was utilized for the trial, and the replicate was regarded as one batch. Twenty ganders and fifty-five dames were used in batch 1 (started on 2011/6/17 and ended on 2012/1/31), thirty ganders and eighty-four dames were used in batch 2 (started on 2012/3/23 and ended on 2012/10/26), and thirty ganders and seventy-two dames were used in batch 3 (started on 2013/3/12 and ended on 2013/12/20). Two hundred and ninety-one geese were randomly assigned to 6 rooms in an environmentally controlled house. They were randomly allotted into one of three monochromatic light treatments: Blue, red, or white. The results showed that there was no significant difference in body weight among the three lighting groups at any point throughout the experimental period. However, compared to the blue light group, significantly more eggs were produced by the red and white light groups (p<0.05). Furthermore, the laying period of the red light group was significantly longer than that of other two groups (p<0.05). In conclusion, our results suggested that red LED-light has the best effect on reproductive performance (i.e. longer laying period and higher total eggs number) at 30 lux light intensity, and is therefore a better choice for the management of breeding geese than blue or white LED-light.

A single nucleotide polymorphism in the promoter region of let-7 family is associated with lung cancer risk in Chinese.

Lung cancer is a complex polygenic disease and many genetic factors are involved in the development of the disease. As one of the most important and widely studied families of microRNA, let-7 appears to play an important role in initiation and progression of lung cancer. Any small changes in miRNA level or its target point can cause significant changes in gene function. In this study, we examined whether a single-nucleotide polymorphism in the promoter region of the let-7 family (rs10877887) is associated with the susceptibility to and prognosis of lung adenocarcinoma cancer. A hospital-based case-control research model was used in our study. The single-nucleotide polymorphism was genotyped in 69 lung cancer patients and 75 healthy controls by direct sequencing. The correlation between rs10877887 genotypes and the susceptibility to lung cancer was evaluated using an unconditional logistic regression model. Populations with the CT+CC genotype had a significantly increased AC risk compared to those with the TT genotype (CT+CC vs TT: P = 0.043, OR = 2.032, 95%CI = 1.018-4.054). Furthermore, the risk effect was greater in subgroups of females over 60 years old (CT+CC vs TT: OR = 6.857, 95%CI = 1.425-33.008, P = 0.012), and the C allele were confirmed to be a risk factor related to lung cancer in these females (P = 0.012). The single-nucleotide polymorphism rs10877887 in the promoter region of the let-7 family was found to be responsible for the susceptibility to lung adenocarcinoma cancer in Chinese individuals. This association was significantly stronger in females who were more than 60 years old.

A study on the relationship between BOD(5) and COD in a coastal seawater environment with a rapid BOD measurement system.

The dual objectives of this study are to: (1) examine the relationship between COD and BOD in seawater environment with a rapid but reliable method for the measurement of BOD in seawater, and (2) establish the relationship model between BOD(5) and COD in the firth of Dongbao River to predict the values of BOD(5). The first objective is met by the successful development of a technique utilizing bacteria-immobilized membrane flow cell for biodegradation process, coupled with fibre optic fluorescence detection for oxygen depletion quantitation. The technique has been applied to coastal seawater samples collected in the coastal area of Shenzhen, China. The BOD(5) and COD values for the samples are acquired and the results show that there is no apparent linear relationship existing between BOD(5) and COD in relatively clean seawater samples away from the shore. However, in estuary water samples containing relatively high concentration of sewage contamination, a linear correlation does exist between BOD(5) and COD. The linear relationship between the two parameters allows for the calculation of BOD(5) values based on COD data which can be measured more readily and precisely.

A study on the relationship between BOD5 and COD in coastal seawater environment with a rapid BOD measurement system.

The dual objectives of this study are to: (1) examine the relationship between COD and BOD in seawater environment with a rapid but reliable method for the measurement of BOD in seawater, and (2) establish the relationship model between BOD(5) and COD in the firth of Dongbao River to predict the values of BOD(5). The first objective is met by the successful development of a technique utilizing bacteria-immobilized membrane flow cell for biodegradation process, coupled with fibre optic fluorescence detection for oxygen depletion quantitation. The technique has been applied to coastal seawater samples collected in the coastal area of Shenzhen, China. The BOD(5) and COD values for the samples are acquired and the results show that there is no apparent linear relationship existing between BOD(5) and COD in relatively clean seawater samples away from the shore. However, in estuary water samples containing relatively high concentration of sewage contamination, a linear correlation does exist between BOD(5) and COD. The linear relationship between the two parameters allows for the calculation of BOD(5) values based on COD data which can be measured more readily and precisely.

Cadmium-induced oxidative cellular damage in human fetal lung fibroblasts (MRC-5 cells).

Epidemiological evidence suggests that cadmium (Cd) exposure causes pulmonary damage such as emphysema and lung cancer. However, relatively little is known about the mechanisms involved in Cd pulmonary toxicity. In the present study, the effects of Cd exposure on human fetal lung fibroblasts (MRC-5 cells) were evaluated by determination of lipid peroxidation, intra-cellular production of reactive oxygen species (ROS), and changes of mitochondrial membrane potential. A time- and dose-dependent increase of both lactate dehydrogenase leakage and malondialdehyde formation was observed in Cd-treated cells. A close correlation between these two events suggests that lipid peroxidation may be one of the main pathways causing its cytotoxicity. It was also noted that Cd-induced cell injury and lipid peroxidation were inhibited by catalase and superoxide dismutase, two antioxidant enzymes. By using the fluorescent probe 2',7'-dichlorofluorescin diacetate, a significant increase of ROS production in Cd-treated MRC-5 cells was detected. The inhibition of dichlorofluorescein fluorescence by catalase, not superoxide dismutase, suggests that hydrogen peroxide is the main ROS involved. Moreover, the significant dose-dependent changes of mitochondrial membrane potential in Cd-treated MRC-5 cells, demonstrated by increased fluorescence of rhodamine 123 examined using a laser-scanning confocal microscope, also indicate the involvement of mitochondrial damage in Cd cytotoxicity. These findings provide in vitro evidence that Cd causes oxidative cellular damage in human fetal lung fibroblasts, which may be closely associated with the pulmonary toxicity of Cd.

Biological monitoring of kidney function among workers occupationally exposed to trichloroethylene.

AIMS: To investigate the nephrotoxic potential of trichloroethylene in a currently exposed population using sensitive urinary markers of kidney toxicity. METHODS: Renal dysfunction was monitored in a cross-sectional study of 70 workers currently exposed to trichloroethylene. An age and sex matched control population of 54 individuals was drawn from hospital and administrative staff. RESULTS: The mean exposure to trichloroethylene, estimated from urinary trichloroacetic acid concentrations, was 32 ppm (range 0.5-252 ppm) with an average duration of exposure of 4.1 years (range 1-20 years). Significant differences between the exposed and control populations were found for nephrotoxicity markers N-acetylglucosaminidase (NAG) and albumin, and for the mode of action marker, formic acid. However, neither NAG nor albumin showed a significant correlation with either the magnitude or duration of exposure to trichloroethylene. There was a significant correlation between urinary formic acid and trichloroacetic acid concentrations. Within the exposed population there were dose dependent increases in urinary methylmalonic acid concentrations and urinary glutathione S-transferase alpha activity. Although still within the control range, these changes were clearly dose dependent and consistent with one of the proposed mechanisms of trichloroethylene induced kidney toxicity. CONCLUSION: Although there was no evidence of kidney toxicity within the population studied, the results suggest that kidney damage could occur at exposure concentrations higher (>250 ppm) than those encountered in this study.

Study on DNA-protein crosslinks induced by chromate and nickel compounds in vivo with 125I-postlabelling assay.

In an attempt to develop biomarkers of chromate and nickel exposure, we have used a rapid, simple and sensitive 125I-postlabelling assay to detect the formation of DNA-protein crosslinks (DPCs) in different tissues from male Sprague-Dawley rats exposed i.p. to potassium chromate (K2CrO4) and nickel chloride (NiCl2). The results demonstrated that 20 h after rats were injected i.p. with these agents, DPCs were observed in WBC, liver and kidney of rats treated with K2CrO4 in doses ranging from 10 to 40 mg/kg body wt. There was a dose-dependent relationship between chromate exposure and DPCs in WBC and liver, but no DPC increase was shown in lung. In the same way, DPCs were found in WBC and lung of rats treated with NiCl2 in doses ranging from 10 to 30 mg/kg in a dose-dependent manner. The formation of DPCs in different tissues was also observed following repeated exposure of rats to K2CrO4 and NiCl2 (10 mg/kg, i.p.) for 3 weeks. These results were similar with the single dose. It is indicated that chromate and nickel compounds possibly cause DNA or protein damage to form DPCs, suggesting DPCs might be useful as a biomarker for quantitative K2CrO4 and NiCl2 exposure and genotoxic lesions. In addition, WBC were shown to be more sensitive to chromate(VI) and nickel(II) induced DPCs than other targets. There were significant correlations between DPCs induced by K2CrO4 in WBC and liver, and by NiCl2 generated DPCs in WBC and lung, indicating that DPCs in WBC may be a good surrogate for some internal organs of humans exposed to chromate(VI) and nickel(II) compounds.

Chromium (VI)-induced production of reactive oxygen species, change of plasma membrane potential and dissipation of mitochondria membrane potential in Chinese hamster lung cell cultures.

OBJECTIVE: To examine whether Reactive Oxygen Species (ROS) is generated, and whether plasma membrane potential and mitochondrial membrane potential are depolarized in Chinese Hamster Lung (CHL) cell lines exposed to Cr (VI). METHODS: CHL cells were incubated with Cr(VI) at 10 mumol/L, 2.5 mumol/L, 0.65 mumol/L for 3 and 6 hours, respectively. The production of ROS was performed by using 2,7-dichlorofluorescin diacetate; The changes in plasma membrane potential were estimated using fluorescent cationic dye DiBAC4; And the changes in mitochondria membrane potential were estimated using fluorescent dye Rhodamine 123. RESULTS: The ROS levels in CHL cells increased in all treated groups compared with the control group (P < 0.01); The plasma membrane potential and mitochondrial membrane potential in CHL cells dissipated after incubated with Cr(VI) at 10 mumol/L for 3 hours and 6 hours (P < 0.01), at 2.5 mumol/L for 6 hours (P < 0.01 or 0.05). CONCLUSION: Cr(VI) causes the dissipation of plasma membrane potential and mitochondrial membrane potential in CHL cell cultures, and Cr(VI)-induced ROS may play a role in the injuries.

DNA strand breaks and poly (ADP-ribose) polymerase activation induced by crystalline nickel subsulfide in MRC-5 lung fibroblast cells.

Nickel compounds are potent carcinogens. Their carcinogenicity is believed to be associated with their solubility and cellular uptake. In the present study, we assessed the in vitro genotoxic effect of a water-insoluble nickel compound, crystalline nickel subsulfide (alpha-Ni3S2) on human embryo lung fibroblast cell line (MRC-5 cells). DNA strand breaks was evaluated using single cell gel electrophoresis, or comet assay. The alpha-Ni3S2 induction of poly (ADP-ribose) polymerase (PADPRP), a nuclear enzyme associated with DNA damage and repair was also studied. Hydrogen peroxide (H2O2) was used as a reference compound. A dose-response relationship was found between alpha-Ni2S2 concentrations (2.5 micrograms/cm2 to 20 micrograms/cm2) and the comet tail length. The increase of PADPRP activity of alpha-Ni2S2 treated MRC-5 cells was also significant and dose-dependent within the concentration range of 2.5 micrograms/ cm2 to 10 micrograms/cm2. Close associations have been found between the comet length and PADPRP level for H2O2 (r = 0.98) and alpha-Ni3S2 (r = 0.97). These results clearly suggest that alpha-Ni3S2 is a potent agent in inducing DNA strand breaks, which may be closely related to its carcinogenic effects. Data from the present study also suggest that both comet assay and PADPRP determination are sensitive techniques for quantitative evaluation of DNA damage induced by nickel compounds.

Upregulation of miR-27a contributes to the malignant transformation of human bronchial epithelial cells induced by SV40 small T antigen.

The introduction of the Simian virus 40 (SV40) early region, the telomerase catalytic subunit (hTERT) and an oncogenic allele of H-Ras directly transforms primary human cells. SV40 small T antigen (ST), which forms a complex with protein phosphatase 2A (PP2A) and inhibits PP2A activity, is believed to have a critical role in the malignant transformation of human cells. Recent evidence has shown that aberrant microRNA (miRNA) expression patterns are correlated with cancer development. Here, we identified miR-27a as a differentially expressed miRNA in SV40 ST-expressing cells. miR-27a is upregulated in SV40 ST-transformed human bronchial epithelial cells (HBERST). Suppression of miR-27a expression in HBERST cells or lung cancer cell lines (NCI-H226 and SK-MES-1) that exhibited high levels of miR-27a expression lead to cell growth arrested in the G(0)-G(1) phase. In addition, suppression of miR-27a in HBERST cells attenuated the capacity of such cells to grow in an anchorage-independent manner. We also found that suppression of the PP2A B56γ expression resulted in upregulation of miR-27a similar to that achieved by the introduction of ST, indicating that dysregulation of miR-27a expression in ST-expressing cells was mediated by the ST-PP2A interaction. Moreover, we discovered that Fbxw7 gene encoding F-box/WD repeat-containing protein 7 was a potential miR-27a target validated by dual-luciferase reporter system analysis. The inverse correlation between miR-27a expression levels and Fbxw7 protein expression was further confirmed in both cell models and human tumor samples. Fbxw7 regulates cell-cycle progression through the ubiquitin-dependent proteolysis of a set of substrates, including c-Myc, c-Jun, cyclin E1 and Notch 1. Thus, promotion of cell growth arising from the suppression of Fbxw7 by miR-27a overexpression might be responsible for the viral oncoprotein ST-induced malignant transformation. These observations demonstrate that miR-27a functions as an oncogene in human tumorigenesis.

α4 is highly expressed in carcinogen-transformed human cells and primary human cancers.

A regulator of the protein phosphatase 2A (PP2A), α4, has been implicated in a variety of functions that regulate many cellular processes. To explore the role of α4 in human cell transformation and tumorigenesis, we show that α4 is highly expressed in human cells transformed by chemical carcinogens including benzo(a)pyrene, aflatoxin B(1), N-methyl-N'-nitro-N-nitrosoguanidine, nickel sulfate and in several hepatic and lung cancer cell lines. In addition, overexpression of α4 was detected in 87.5% (74/80) of primary hepatocellular carcinomas, 84.0% (21/25) of primary lung cancers and 81.8% (9/11) of primary breast cancers, indicating that α4 is ubiquitously highly expressed in human cancer. Functional studies revealed that elevated α4 expression results in an increase in cell proliferation, promotion of cell survival and decreased PP2A-attributable activity. Importantly, ectopic expression of α4 permits non-transformed human embryonic kidney cells (HEKTER) and L02R cells to form tumors in immunodeficient mice. Furthermore, we show that the highly expressed α4 in transformed cells or human tumors is not regulated by DNA hypomethylation. A microRNA, miR-34b, that suppresses the expression of α4 through specific binding to the 3'-untranslated region of α4 is downregulated in transformed or human lung tumors. Taken together, these observations identify that α4 possesses an oncogenic function. Reduction of PP2A activity due to an enhanced α4-PP2A interaction contributes directly to chemical carcinogen-induced tumorigenesis.

Isolation and characterization of viruses related to the SARS coronavirus from animals in southern China.

A novel coronavirus (SCoV) is the etiological agent of severe acute respiratory syndrome (SARS). SCoV-like viruses were isolated from Himalayan palm civets found in a live-animal market in Guangdong, China. Evidence of virus infection was also detected in other animals (including a raccoon dog, Nyctereutes procyonoides) and in humans working at the same market. All the animal isolates retain a 29-nucleotide sequence that is not found in most human isolates. The detection of SCoV-like viruses in small, live wild mammals in a retail market indicates a route of interspecies transmission, although the natural reservoir is not known.