Expression of indoleamine 2,3-dioxygenase in pregnant mice correlates with CD4+CD25+Foxp3+ T regulatory cells.

OBJECTIVE: Indoleamine 2,3-dioxygenase (IDO) initiated tryptophan degradation in the placenta has a role in the prevention of allogeneic fetus rejection by T-cells. The present study aimed to investigate the relationship between IDO and CD4+CD25+Foxp3+ T cells in pregnant mice. MATERIALS AND METHODS: The percentage of CD4+CD25+Foxp3+ T cells in peripheral blood mononuclear cells (PBMC) and IDO mRNA levels were detected in pregnant mice. The non-pregnant mice were used as control in this study. To confirm the effect of IDO, 1-methyl-trytophan (IDO inhibitor) was used in this study. RESULTS: The percentage of CD4+CD25+Foxp3+T cells in PBMC in pregnant mice was significantly higher than this in non-pregnant mice controls from day-6 to the end of the study (p<0.05). IDO mRNA levels in PBMC also markedly increased after pregnancy. The upregulation of IDO expression reached a maximum at day 18 after pregnancy (p<0.05). Compared to the pregnant group, the inhibitor could significantly decrease the IDO expression and Treg percentage (p<0.05). There was a positive association between IDO mRNA and CD4+CD25+Foxp3+ T cells percentage. CONCLUSIONS: The results suggested IDO might play a role in the generation of CD4+CD25+Foxp3+ T cells in pregnant mice.

Molecular cloning, sequence analysis, prokaryotic expression, and function prediction of foot-specific peroxidase in Hydra magnipapillata Chinese strain.

The cDNA sequence of foot-specific peroxidase PPOD1 from the Chinese strain of Hydra magnipapillata was cloned by reverse transcription-polymerase chain reaction. The cDNA sequence contained a coding region with an 873-bp open reading frame, a 31-bp 5'-untranslated region, and a 36-bp 3'-untranslated region. The structure prediction results showed that PPOD1 contains 10.34% of α-helix, 38.62% of extended strand, 12.41% of ß-turn, and 38.62% of random coil. The structural core was α-helix at the N terminus. The GenBank protein blast server showed that PPOD1 contains 2 fascin-like domains. In addition, high-level PPOD1 activity was only present in the ectodermal epithelial cells located on the edge of the adhesive face of the basal disc, and that these cells extended lamellipodia and filopodia when the basal disc was tightly attached to a glass slide. The fascin-like domains of Hydra PPOD1 might contribute to the bundling of the actin filament of these cells, and hence, the formation of filopodia. In conclusion, these cells might play an important role in strengthening the adsorbability of the basal disc to substrates.

Association of CYP1B1 gene polymorphisms and the positive expression of estrogen alpha and estrogen beta with endometrial cancer risk.

To investigate the relationship between the CYP1B1 L432V polymorphism, ERalpha and ERbeta positivities and the incidence of endometrial cancer. The relationship between CYP1B1 L432V polymorphism, ERalpha and ERbeta positivities and endometrial cancer was investigated using the allele-specific polymerase chain reaction method to analyze gene polymorphism in exon 3 codon 432 (C-G) of CYP1B1. Our results are as follows: in endometrial cancer cases the prevalence rates of CYP1B1 L432V genotypes C/C, C/G, and G/G were 47.2%, 36.1%, and 16.7%, respectively, and 68.8%, 23.8% and 7.5% in the control group, respectively. The frequencies of CYP1B1 C and G alleles were 65.3% and 34.7% in endometrial cancer patients and 80.6% and 19.4% in the control group. A significant difference was found in the genotype distributions or allele frequencies of CYP1B1 L432V polymorphism between the two groups (p < 0.05). Compared with wild-type C/C, the susceptibility of endometrial cancer with homozygotic mutation G/G and heterozygotic mutation C/G increased by 3.235 (95%CI 1.111-9.425) and 2.214 (95% CI 1.067-4.593). Moveover, the positive expression of ERalpha in genotypes G/G and C/G was higher than in the wild genotype C/C (p < 0.05). In conclusion, allelic polymorphism of CYP1B1 L432V increases the risk of endometrial cancer and has a positive correlation with ERalpha expression.

Interruption of the phosphoglucose isomerase gene results in glucose auxotrophy in Mycobacterium smegmatis.

Two glycerol utilization mutants of Mycobacterium smegmatis that were unable to utilize most carbon sources except glucose were isolated. Supplementation of these media with small amounts of glucose restored growth in the mutants; these strains are therefore glucose auxotrophs. The mutant phenotype is complemented by the gene encoding phosphoglucose isomerase (pgi), and direct measurement of enzyme activities in the mutants suggests that this gene product is absent in the auxotrophic strains. Mapping of the mutant allele by Southern analysis demonstrates the presence of a 1-kb deletion extending into the coding sequence of pgi. The possible roles of phosphoglucose isomerase in mycobacterial cell wall synthesis and metabolic regulation are discussed.

[Use of polymerase chain reaction with L1 consensus primer for detection of HPV16 and HPV18 in cervical cancer tissues].

Biopsy materials of cervical carcinoma including 20 cervical adenocarcinomas and 20 squamous cell carcinomas were collected. A rapid method for determining HPV type was developed, based on DdeI restriction enzymes analysis within the L1 region of HPV, amplified by PCR using consensus primers. The results indicated that HPV type 16 was detected more often in squamous cell carcinomas than in adenocarcinomas (80% vs 15%, P < 0.001), conversely, HPV type 18 was detected significantly more often in adenocarcinoma tissues (45% vs 5%, P < 0.001). These differences may reflect the presence of different virus receptors in cancer cells with different morphologic potential, or, they may indicate that the specific HPV infection actually plays a direct role in the process of carcinogenesis.

Stable preparation of aldose reductase isoenzymes from human placenta.

An efficient, large-scale purification has been achieved for two aldose reductase isoenzymes from human placenta in stable form. The procedure included ammonium sulfate fractionation (45-75%), followed by chromatographies on Matrex Red A, DE-52 cellulose, and Matrex Orange A. The preparations were stable for at least 3 months at 3 degrees C. IC50 values toward sorbinil were similar to those reported for crude or partially purified enzymes, indicating that they retained native structures during the purification steps. The molecular weights of purified GAR1 and GAR2, named according to their order of elution with a salt gradient from a Matrex Red A column, were 36,600 and 40,300, respectively. Kinetic studies indicate that GAR1 belongs to an aldose reductase (a low-Km form) and GAR2 to an aldehyde reductase (a high-Km form). GAR2, an aldehyde reductase, was also active in the reduction of D-glucose, with an apparent Km comparable to that of GAR1 but with a Vmax only 14% that of GAR1.