Treg Cell Evaluation in Patients with Acquired Immune Deficiency Syndrome with Poor Immune Reconstitution and Human Immunodeficiency Virus-Infected Treg Cell Prevention by Polymeric Nanoparticle Drug Delivery System.

To better deliver antiretroviral drugs for treating patients with acquired immune deficiency syndrome (AIDS) with poor immune reconstitution, a novel nanopole capsule was designed in this study. Forty-eight patients with AIDS with poor immune reconstitution were chosen as subjects to test their immune state. CD4+ T and Regulatory T cells (Treg) infected with HIV were cultured to test polyethyleneimine (PEI) and polychitosan (PC) drug delivery system efficiency. The infiltration efficiency test was performed to study the drug delivery efficiency of the delivery systems, and the cell numbers of CD4+ T and Treg cells infected with HIV were calculated to evaluate the therapeutic effect. The results showed that patients with AIDS with poor immune reconstitution had lower CD4+ T cell count and higher Treg cell count. Furthermore, the infiltration efficiency of the PC drug delivery system was higher than that of the PEI drug delivery system, and the therapy efficiency of antiretroviral drugs was greatly improved in the PC group. Additionally, the improvement of CD4+ T and Treg cells damaged by HIV was greater in the PC group. Sequentially, the PC system can better deliver and release loaded antiretroviral drugs and may be a better choice for treating patients with AIDS with poor immune reconstitution in the future.

Gene systems network inferred from expression profiles in hepatocellular carcinogenesis by graphical Gaussian model.

Hepatocellular carcinoma (HCC) in a liver with advanced-stage chronic hepatitis C (CHC) is induced by hepatitis C virus, which chronically infects about 170 million people worldwide. To elucidate the associations between gene groups in hepatocellular carcinogenesis, we analyzed the profiles of the genes characteristically expressed in the CHC and HCC cell stages by a statistical method for inferring the network between gene systems based on the graphical Gaussian model. A systematic evaluation of the inferred network in terms of the biological knowledge revealed that the inferred network was strongly involved in the known gene-gene interactions with high significance (P < 10(-4)), and that the clusters characterized by different cancer-related responses were associated with those of the gene groups related to metabolic pathways and morphological events. Although some relationships in the network remain to be interpreted, the analyses revealed a snapshot of the orchestrated expression of cancer-related groups and some pathways related with metabolisms and morphological events in hepatocellular carcinogenesis, and thus provide possible clues on the disease mechanism and insights that address the gap between molecular and clinical assessments.

Apoptosis and its pathway in early post-implantation embryos of diabetic rats.

It has been reported that diabetes-induced inappropriate apoptosis in embryos during neurulation may be one of the mechanisms leading to neural tube defects. We studied apoptosis and the apoptotic pathway occurring in early post-implantation period embryos of non-diabetic and streptozotocin (STZ)-induced diabetic rats. In quantitative RT-PCR, bax mRNA was constantly expressed to similar degree in embryos of non-diabetic and diabetic rats, while the expression of bcl-2 mRNA was significantly decreased in diabetic rat embryos compared to non-diabetic rat embryos. The increased number of terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL)-positive cells occurred selectively in the primitive brains of diabetic rat embryos compared to non-diabetic rat embryos. Immunohistochemical studies revealed that, in mirror sections, the staining of Bax and activated caspase-3 were observed in the TUNEL-positive cell area, but the expression of Bcl-2 in these apoptotic cells was generally too low to be detected. These results suggest that a Bax-regulated mitochondrial cytochrome c-mediated caspase-3 activation pathway might be involved in the diabetic embryopathy.

Association between IL-18 gene promoter polymorphisms and CTLA-4 gene 49A/G polymorphism in Japanese patients with type 1 diabetes.

Interleukin-18 (IL-18) is a potent proinflammatory cytokine which is strongly associated with the development of diabetes in NOD mice. To test the putative involvement of IL-18 gene polymorphism in predisposition to human type 1 diabetes, the SNPs at position -607 (C/A) and -137 (G/C) in the promoter region of IL-18 gene were analyzed by sequence-specific PCR in 116 patients with type 1 diabetes and 114 normal controls. A linkage disequilibrium found only three of the four possible haplotypes defined by these SNPs. The distribution of the IL-18 gene genotypes at position -607 was significantly different between patients with type 1 diabetes and normal controls (P=0.023). Furthermore, there was a significant increase in haplotype 1 (-607C/-137G) in the patients compared with controls (P=0.006). The association study of the susceptible CTLA-4 genotype (GG at nucleotide position 49 in exon 1) or HLA-DR4-DQB1*0401 and type 1 diabetes showed that the predisposing IL-18 gene haplotype modulates the risk on CTLA-4 GG genotype, but not on HLA-DR4-DQB1*0401 haplotype. Among subjects carrying the CTLA-4 GG genotype, the frequency of IL-18 haplotype 1 in patients with type 1 diabetes was significantly higher than that in controls (91% vs. 71%, P=0.012). However, IL-18 haplotype 1 was not frequent in patients who do not exhibit the CTLA-4 high-risk genotype. These results suggest that the IL-18 gene polymorphism is associated with a type 1 diabetes susceptibility, and there might be a gene-gene interaction between IL-18 gene with susceptible CTLA-4 gene.

Restricted expression and photic induction of a novel mouse regulatory factor X4 transcript in the suprachiasmatic nucleus.

The regulatory factor X (RFX) family of transcription factors is characterized by a unique and highly conserved 76-amino acid residue DNA-binding domain. Mammals have five RFX genes, but the physiological functions of their products are unknown, with the exception of RFX5. Here a mouse RFX4 transcript was identified that encodes a peptide of 735 amino acids, including the DNA-binding domain. Its expression was localized in the suprachiasmatic nucleus, the central pacemaker site of the circadian clock. Also, light exposure was found to induce its gene expression in a subjective night-specific manner. Polyclonal antibodies were prepared, and an 80-kDa band was detected in the suprachiasmatic nucleus by Western hybridization. A histochemical study showed a localization of the products in the nucleus. This is the first report on mouse RFX4, which contains the RFX DNA-binding motif. Our investigation may provide clues to the physiological function of RFX4.

In vivo expression of B:9-23 peptide/I-A(g7) complex may abrogate the inhibition of diabetes induced by RGD-fiber-mutant adenovirus in NOD mice.

Insulin B chain peptide B:9-23 given to NOD mice decreases the development of diabetes, and phase II trials of an altered peptide ligand of B:9-23 are under way in humans. We have created a gene for the NOD MHC class II beta chain, covalently linked to the B:9-23 peptide. B lymphoma cells transfected with the gene stimulated NOD islet-derived B:9-23 reactive T cell clones in vitro. In this study, we generated an RGD-fiber-mutant adenovirus vector encoding the covalent B:9-23 peptide/I-A(g7) gene (Ad-RGD-B:9-23) to test whether in vivo expression of the gene could protect NOD mice from diabetes. NOD female mice were injected intramuscularly with 5 x 10(8) PFU of Ad-RGD-B:9-23 and empty RGD-adenovirus vector. A single administration of the empty vector did not alter the expression of insulin autoantibodies, but delayed the onset of diabetes in NOD mice. In contrast, Ad-RGD-B:9-23 immunization induced an early expression of insulin autoantibodies, but did not change the disease occurrence compared to control NOD mice. Our results suggest that adenovirus infection could confer protection from diabetes in NOD mice. The in vivo expression of covalent B:9-23 peptide/class II complex by adenovirus gene transfer might activate anti-insulin autoimmunity, resulting in abrogation of the inhibition of diabetes induced by an RGD-fiber-mutant adenovirus vector.

Apoptosis in normal rat embryo tissues during early organogenesis: the possible involvement of Bax and Bcl-2.

Apoptosis commonly occurs in a variety of developmental processes in mammals. In this study, we investigated the relationship between apoptosis and the expression of both Bax and Bcl-2 during the early organogenesis period (9.5-11.5 days of gestation) of rat embryos. Apoptotic cells detected by the terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) method were extremely abundant in the foregut diverticulum at 9.5 days of gestation, while they largely disappeared at 10.5 and 11.5 days of gestation, although they were detected in newly formed mid- and hindgut diverticulum at these times. Real-time RT-PCR analysis of whole embryos revealed that the expression of bax mRNA was constant at days 9.5 to 11.5, while the expression of bcl-2 mRNA gradually increased. Immunohistochemical studies of Bax and Bcl-2 expression revealed that these apoptotic cells were exactly positive to Bax in mirror sections, while their expression of Bcl-2 was generally too low to be detected. A disappearance of apoptotic cells was associated with strong Bcl-2 expression in the foregut diverticulum at 10.5 and 11.5 days of gestation. It was similarly observed that apoptotic cells detected in the cardiogenic area at 9.5 days of gestation disappeared with the formation of the primitive heart tube--accompanied by a strong expression of both Bcl-2 and Bax--in the developmental process of the primitive heart. Apoptotic cells were also observed in the primitive brain vesicle, optic vesicle, otic vesicle, and thyroid primordium at 10.5 and 11.5 days of gestation during the developmental process, with a strong expression of Bax. These results indicate that the Bax and Bcl-2 may be important in regulating the induction of embryonic cell apoptosis during early organogenesis.