Association of interleukin 22 gene polymorphisms and serum IL-22 level with risk of systemic lupus erythematosus in a Chinese population.

The aim of this study was to investigate the association between the single-nucleotide polymorphisms (SNPs) of the interleukin 22 (IL-22) gene and systemic lupus erythematosus (SLE) in a Chinese population. Three IL-22 SNPs (rs2227485, rs2227513 and rs2227491) were genotyped using SNaPshot SNP genotyping assays and identified by sequencing in 314 SLE patients and 411 healthy controls. The IL-22 level of serum was assessed by enzyme-linked immunosorbent assay (ELISA) kits. Data were analysed by spss version 17.0 software. We found that rs2227513 was associated with an increased risk of SLE [AG versus AA: adjusted odds ratio (aOR) = 2·24, 95% confidence interval (CI) = 1·22-4·12, P = 0·010; G versus· A: adjusted OR = 2·18, 95% CI = 1·20-3·97, P = 0·011]. Further analysis in patients with SLE showed that the AG genotype and G allele were associated with an increased risk of renal disorder in SLE (G versus A: aOR = 3·09, 95% CI = 1·30-7·33, P = 0·011; AG versus· AA: aOR = 3·25, 95% CI = 1·35-7·85, P = 0·009). In addition, the concentration of IL-22 was significantly lower in the rs2227513 AG genotype compared with AA genotype (P = 0·028). These results suggest that rs2227513 polymorphism might contribute to SLE susceptibility, probably by decreasing the expression of IL-22.

[Association of RTN4 gene rs2864052 and rs6545468 with the susceptibility of nasopharyngeal carcinoma in Guangxi Zhuang population].

Objective:To study the relationship of the polymorphism of RTN4 gene rs2864052 and rs6545468 and haplotype with the susceptibility of nasopharyngeal carcinoma in Guangxi Zhuang population. Method:The polymorphism of Nogo gene (rs2864052,rs6545468) and haplotype were analyzed using the method of single-base extension PCR and DNA sequencing in 282 cases of nasopharyngeal carcinoma (NPC) and 199 healthy persons (control group) in Guangxi Zhuang Autonomous Region. Result:There were no differences between the NPC's patients and controls in the genotype and allele frequencies of RTN4 gene rs2864052 site,or rs6545468 site. The frequency of AG haplotype in the NPC's patients was significantly lower than in the controls(P=0.004, OR=0.14,95%CI=0.31-0.68). Conclusion:The haplotype AG of RTN4 gene rs2864052 and rs6545468 sites may reduce the risk of nasopharyngeal carcinoma in Guangxi Zhuang population.

Development of test systems for the detection of compounds that prevent the genotoxic effects of heterocyclic aromatic amines: preliminary results with constituents of cruciferous vegetables and other dietary constituents.

Over the past decades, strong efforts have been made to identify dietary constituents that protect against the genotoxic effects of heterocyclic aromatic amines (HAAs). However, most of the methods that have been used, in particular in vitro assays that require the addition of exogenous enzyme homogenates, have only a limited predictive value because important protective mechanisms are not adequately represented and may give misleading results. Therefore, we attempted to develop improved test systems, namely assays, with human hepatoma cells and single-cell gel electrophoresis (SCGE) tests with rats. Genotoxicity tests with human derived Hep G2 cells reflect the genotoxic effects of HAAs better than other in vitro systems. They also enable the detection of protective effects since the human derived hepatoma cells possess phase I and phase II enzymes that are involved in the activation/ detoxification of the amines. The most appropriate endpoint for experiments with Hep G2 cells appears to be micronucleus induction, but protocols for other endpoints are available as well. The second promising model is the SCGE ("comet") assay with rats that was used successfully to measure protective effects of constituents of cruciferous vegetables against 2-amino-3-methylimidazo[4,5-flquinoline (IQ) in the liver and in the colon mucosa. The present study describes the experimental design of the new approaches, as well as results obtained with various dietary constituents.

LIF upregulates poFUT1 expression and promotes trophoblast cell migration and invasion at the fetal-maternal interface.

Trophoblast cell migration and invasion are crucial for the establishment of a successful pregnancy. Protein O-fucosyltransferases, such as poFUT1 and poFUT2, catalyze the O-fucosylation of proteins and have important roles in embryonic development. Leukemia inhibitory factor (LIF) is a critical cytokine in the regulation of embryonic development and implantation. However, the exact roles of poFUTs in embryo migration and invasion and the effects of LIF on the expression of poFUTs have not been studied in detail. In the current study, we showed that poFUT1 and LIF were highly expressed in human trophoblast cells and in the serum of women during the first trimester of a normal pregnancy. However, in patients with threatened abortion, poFUT1 and LIF levels were found to be reduced. There were no significant differences in the expression levels of poFUT2 between the two groups. The migration and invasion potential of trophoblasts in an explant culture and in an in vitro implantation model was decreased or increased upon altering poFUT1 expression levels by siRNA or cDNA transfection. Our results also revealed that LIF upregulated the expression of poFUT1. The upregulation of poFUT1 by LIF promoted trophoblast cell migration and invasion at the fetal-maternal interface by activating the PI3K/Akt signaling pathway. Taken together, these study findings suggest that poFUT1 may be used as a marker of embryo implantation.

Effect of three tumour promoters on the stability of hepatocyte cultures and apoptosis after transforming growth factor-beta 1.

Tumour promoters like the anti-androgen cyproterone acetate (CPA), the peroxisome proliferator nafenopin (NAF) and phenobarbital (PB) stimulate liver growth in rodents. Transforming growth factor-beta 1 (TGF-beta 1) is expressed in livers after treatment with CPA (Oberhammer et al., submitted) and some peroxisome proliferators. In this paper we describe the influence of CPA, NAF and PB on the stability of hepatocyte cultures and induction of apoptosis by TGF-beta 1. All three tumour promoters had a stabilizing effect on confluent monolayers of hepatocytes, partially preventing the usually occurring dedifferentiation and detachment processes. CPA on its own was able to induce apoptosis at the high dose of 10 microM. No induction of apoptosis could be observed after PB and NAF. At any dose above 0.01 microM CPA enhanced TGF-beta 1-induced apoptosis (5.8-fold increase with 10 microM CPA). Thus the combination of 10 microM CPA and 1 ng/ml TGF-beta 1 induced apoptosis in 90% of the plated hepatocytes. At a high dose (10 microM) NAF produced a 35% reduction in apoptosis induced by TGF-beta 1, in parallel with a stabilizing effect on cell number. PB did not affect the rate of apoptosis induced by TGF-beta 1. As demonstrated by immunohistochemical detection of PCNA, TGF-beta 1 prevented induction of PCNA by epidermal growth factor (EGF). No induction of PCNA was observable in CPA-treated cultures. In untreated and EGF-treated cultures TGF-beta 1 was able to induce apoptosis to the same extent within 30 h. In CPA-treated cultures this period was shortened to 12 h. Thus CPA shortens the lag phase of induction of apoptosis by shifting hepatocytes to a point before S phase, where they are highly susceptible to TGF-beta 1-induced apoptosis.

[Studies on transgenic tobacco plants expressing two kinds of insect resistant genes].

A synthetic Bt cry1Ac gene fussed with a secretary signal coding sequences at 5' end and a modified gna gene were used to construct a plant expression vector pBSGS1M+ and this vector was transferred into tobacco (Nicotiana tabacum L.) by Agrobacterium-mediated transformation method. Results of PCR, Southern blot and Slot blot analysis indicated that both the chimeric Bt cry1Ac and gna genes were integrated into the genomes of transformed plants. Western blot analysis indicated that at least the cry1Ac protein was produced in transgenic plants. Upon insect bioassay using cotton bollworm (Heliothis armigera Hubner), the mortality of insect larvae on 60% regenerated plants reached 100% in 5 days post infestation and the growth of the survived larvae was seriously inhibited; The results from insect bioassay with peach aphid (Myzus persicae) showed that the transgenic plants were aphid-resistant, evidenced by a 50%-60% reduction in aphid population density, even over 80% for some individual transgenic plants. These results reflect that the modification of the two insect resistant genes and construction of the expression vector are correct and could be valuable for later application in crop breeding for insect resistance.

Intestinal microflora plays a crucial role in the genotoxicity of the cooked food mutagen 2-amino-3-methylimidazo [4,5-f]quinoline.

We investigated the impact of the intestinal microflora on the genotoxicity of 2-amino-3-methylimidazo[4,5-f] quinoline (IQ), a mutagenic/carcinogenic heterocyclic amine commonly found in fried meats and fish. In parallel, we also examined the effect of the microflora on the protective effect of glucotropaeolin (GT), a glucosinolate contained in cruciferous vegetables, towards IQ-induced genotoxic effect. Conventional (NF), human flora associated (HFA) and germ free (GF) rats were treated either with 90 mg/kg IQ alone, 150 mg/kg GT alone or a combination of the two by gavage and DNA damage was determined in liver and colon cells using the alkaline single cell gel electrophoresis (SCGE) or comet assay. IQ caused a significant effect in both organs of all groups. However, DNA damage was most pronounced in NF animals. In colon cells, DNA migration was 6-fold more in IQ-exposed rats as compared with untreated controls. The effect measured with liver cells was similar. In comparison to NF rats, in HFA rats, tail length of the comets was 22 and 53% lower in liver and colon cells, respectively. Significantly weaker effects were seen in GF animals (66 and 75% lower damage in hepatocytes and colonocytes, respectively, than in NF animals). Pretreatment with GT led to a complete reduction of IQ-induced DNA damage regardless of the microbial status of the animals. In addition, a moderate decrease in spontaneous DNA damage was seen in animals that received GT alone. Our results show that the microflora has a strong impact on the genotoxic effects of IQ. We conclude that the alkaline SCGE assay with rats harbouring different flora opens new possibilities to investigate the role of intestinal bacteria on health risks caused by dietary carcinogens.

A novel method for detecting apoptosis shows that hepatocytes undergo a time dependent increase in DNA cleavage and chromatin condensation which is augmented after TGF-beta 1 treatment.

This study describes a new method for quantitating apoptosis in hepatocyte monolayers in which nuclei were isolated from the cells and DNA strand breaks detected by in situ end-labeling and flow cytometry. Most (97%) nuclei from untreated hepatocytes had low end-labelling and were derived from non-apoptotic cells. Approximately 2-3% of the nuclei had high end-labelling and originated from apoptotic hepatocytes. The numbers of these nuclei increased linearly from 3 to 85% between 0 and 48 h after treatment with transforming growth factor-beta 1 (TGF-beta 1). However, a morphological assessment of apoptosis with Hoechst H33258 showed that the proportion of apoptotic nuclei plateaued at 18-19% between 24 and 48 h after TGF-beta 1 treatment. Thus, the in situ end-labeling technique also detected DNA cleavage in nuclei which did not have an obvious apoptotic morphology. Confocal microscopy of low and high end-labelled nuclei which had been separated by fluorescent cell sorting showed that nuclei with high levels of end-labeling exhibited a wide diversity of morphologies. These included nuclei with little or no chromatin condensation and nuclei with characteristic apoptotic morphology. In addition, nuclei from untreated hepatocytes contained low levels of DNA cleavage, which were localized in areas of condensed chromatin and increased according to the time in culture. Thus, hepatocytes undergo a progressive and cumulative process of DNA cleavage/chromatin condensation which is markedly enhanced by TGF-beta 1.