Objective To explore the genetic damages of people exposed to electronic waste from three villages that have been processing the electronic waste collectively in county of Tianjin, and to analyze the damaging effects of the electronic pollutants on the genetic substances. Methods The blood samples of 1 256 villagers from this region were collected to do chromosomal karyotype analysis, from which 171 samples were randomly chosen to do micronucleus test, 12 samples were chosen to do comet assay experiments. 60 villagers living far from the electronic waste processing regions were chosen as the control group. Chi-Square test and T-test were employed to do statistic analysis. Results The chromosomal structural aberration in the research group was 6.23%, and the number aberration was 0.29%. Both of chromosomal conjoint and satellite conjoint were higher compared with the control (P

Dendritic cells (DCs) are potent antigen presenting cells with dual functions in immune response or im-mune tolerance. Regulatory dendritic cells (DCreg) have negative immunomodulation effects. They can inhibit T cell stimula-tory activity and induce immune tolerance. DCreg can be generated by the induction of drugs, cytokines and cell microenvi-ronment with different mechanisms. DCreg induced tolerance is of great clinical significance in organ transplantation and au-toimmune disease. Recently, several clinical trials have been conducted in type 1 diabetes and rheumatoid arthritis, with re-sults that emphasize the tolerance and safety of DCreg therapy. In this review, we will focus on the strategies for generating DCreg and the clinical application of immune tolerance.

Objective To explore the immunomodulation property of bone marrow-derived mesenchymal stem cells (BMSCs) from Sprague-Dawley (SD) rats after they are isolated, cultured and identified by surface marker and differentiation potential examination. Methods BMSCs were isolated from femur and tibia of SD rats and passaged by trypsinization. The surface markers of the 3rd passage BMSCs were detected by flow cytometry and the capacity of their adipocyte and cartilage differentiation were examined. In order to explore the immunomodulation property of BMSCs, allogeneic spleen T cells of Wi?star rats were co-cultured with BMSCs through either cell-to-cell contact or transwell, then its effect on the T cell subsets and related mechanism was also examined. Results BMSCs were mainly spindle-shaped in culture. Surface marker detec?tion showed that BMSCs expressed high levels of CD29, CD44 and CD90 but no CD34 nor CD45 at the third generation. Un?der specific condition, BMSCs could differentiate into adipocytes and chondrocytes. The CD8+effector T cells (Teffs) decreas?es effectively and the CD4+CD25+regulatory T cells (Tregs) increased remarkably when BMSCs were co-cultured with allo?geneic spleen T cells for 48 hours. The expressions of IL-10 and TGF-β1 of BMSCs significantly increased after co-culture with T cells, and this effect was more obvious in cell-to-cell contact group. Conclusion The immunomodulation property of BMSCs were presumably function through cell-to-cell contacts and cytokine secretion.

Objective To investigate the protein markers that specifically expressed in patients with acute rejection (ACR) after liver transplantation, and to explore preliminarily the mechanisms. Methods Serum samples from three patients with pathologically confirmed ACR after liver transplantation in Tianjin First Central Hospital were collected as ACR group. Three serum samples from patients with normal liver function indicators after liver transplantation were collected as No-ACR group. And six serum samples from healthy examination were mixed with equal amount as healthy control group. Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) was employed to separate, screen and identify the differentially expressed proteins between three groups. KEGG and STRING software were applied to deeply analyze the data of three groups. Results A total of 88 differentially expressed proteins were found between ACR group and healthy control group. There were 39 differentially expressed proteins between No-ACR group and healthy control group. Ten differentially expressed proteins were acquired between ACR group and No-ACR group. Comparing 88 and 10 differentially expressed proteins, 9 proteins were the same. Among 88 differentially expressed proteins, 30 of them showed a direct interaction, and can be positioned in 13 signaling pathways based on KEGG and STRING software. Fourteen (46.67%) of the 30 proteins were located in the complement and coagulation cascade pathway. Among 39 differentially expressed proteins, which were detected between No-ACR group and control group, 10 proteins showed a direct interaction including 9 proteins concentrated in the complement and coagulation cascade pathway. Conclusion By proteomic analysis, nine differentially expressed proteins are obtained, which may be regarded as the candidate bio-markers for ACR early diagnosis after liver transplantation. The complement and coagulation cascades system is significantly adjusted after liver transplantation, indicating this pathway plays an important role in the occurrence of ACR.

Objective To explore the effects of traditional Chinese formula Danchaiheji on the differentiation of regula?tory dendritic cells (DCs) and the underlying mechanism. Methods The rat blood serums with or without the formula Dan?chaiheji were prepared. The peripheral blood mononuclear cells were separated from the peripheral venous blood of healthy donors. CD14+monocytes were isolated using CD14+magnetic beads and cultured for 5-7 days to obtain immature dendritic cells (imDCs). Then the cells was divided into control group and Danchaiheji containing rat serum group. Control group was divided into two subgroups (containing LPS and without LPS). Danchaiheji containing rat serum group was also divided into two subgroups (containing LPS and without LPS). The surface markers CD86, CD11b and HLA-DR of DCs were detected by flow cytometry. The level of IL-10 was determined by enzyme-linked immunosorbent assays (ELISA). The proliferation of al?logeneic T-cells was detected by flow cytometry and the expression level of indoleamine 2,3-dioxygenase (IDO) was deter?mined using quantitative real-time PCR. Results DCs treated with the formula Danchaiheji exhibited high CD11b and low CD86 and HLA-DR expression levels as well as promoted the secretion of IL-10. In addition, the drug could inhibit the pro?motion of DCs on the proliferation of T cells, which was associated with the up-regulation of IDO expression. Conclusion The traditional Chinese formula Danchaiheji can induce the differentiation of DCs into regulatory DCs and play a role in in?hibitory effect on immune function.

Objective:To assess the expression levels of CD95 and OX40 ligand(OX40L)messenger RNA(mRNA)in hepatocellular carcinoma(HCC),and their clinical values thereof.Methods:The lymphocytes of research objects were mixed with corresponding fluorescence labeled monoclonal antibody(McAb);CD95 expression by CD3 positive T ceils was quantitatively measured by dual color flow cytometry.The expression of OX40L mRNA was detected in peripheral blood mononuclear cells by fluorescence quantitative reverse transcription polymerase chain reaction(FQ-RT-PCR).Results:The CD95 expression by CD3 positive T cells was significantly higher in patients with HCC(34±20)% compared with that of normal controls (20±7)%(t=2.960,P ＜ 0.01).The expression level of OX40L mRNA was significantly decreased in patients with HCC compared with that of normal controls(t=2.302,P ＜ 0.05).Conclusion:These results suggest that abnormal expressions of CD95 and OX40L play crucial roles in peripheral blood of HCC patients at the stage of human hepatocareinogenesis.

Objective To analyse the change of DNA methylation with 5-Azac injection in acute graft-versus-host dis- ease (aGVHD) mouse model, which received allogeneic bone marrow transplantation, and explore the immunomodulatory ef-fects of 5-Azac. Methods Male C57BL/6 (H-2b)and female BALB/c (H-2d) mice were selected as donor and recipient of complete allotransplantation. BABL/c mice were divided into two groups, transplantation control group and 5-Azac experi-mental group. At 1-7, 14, 21 and 28-day after transplantion, 5-Azac 0.25 mg/kg (0.3 mL/time) was injected by tail vein in experimental group, while the control group were injected with sterile water 0.3 mL/time. Peripheral blood DNA samples were collected from three control mice and three experimental mice, then mixed with equal amount respectively. The MeDIP-seq method was selected to detect methylation changes in mice, and the differential DNA methylation in the biological path-ways was analyzed. Results The survival time was prolonged, and the rejection reaction was decreased in 5-Azac experi-mental group, which suggested immune hyporesponsiveness post aGVHD. The MeDIP-seq result showed that 369 different DNA methylation located in the promoter regions, including 239 up-regulated genes and 130 down-regulated genes. There were 184 differential DNA methylation genes located in the exon regions, including 113 up-regulated genes and 71 down-regulated genes. Differential DNA methylation genes involved in 10 immunological signaling pathways, respectively. Among them, TGF-β, GSK-3β, SYK, PI3K, NFAT, CD28 andα4β7 were closely related to the development of aGVHD. Conclu-sion 5-Azac can effectively induce immune hyporesponsiveness post aGVHD by changing the gene methylation status.

Objective To investigate COL1A1 gene mutation by PCR-high resolution melting (PCR-HRM) and an-alyze the correlation between genotype and clinical phenotype in a child (proband) with osteogenesis imperfecta (OI). Methods The family history of OI pedigree along with the clinical data was collected. Blood samples from the proband and his family members, as well as 50 normal controls, were collected. The mutation of COL1A1 gene was screened using PCR-HRM and validated by the gene sequence. Results The detection of PCR-HRM showed the abnormal result of COL1A1 17 exon in proband with a lower melting temperature (Tm) value than that of normal controls by 0.4℃. There were signifi-cant differences in the standardization melting curve and the different melting curve between the proband and the normal controls. The sequencing result was c.1138G>A, which meant that cDNA of 1138 base G mutation into A. The mutations transformed the amino acid glycine into a serine at amino acid 380(P. Gly 380 Ser), which resulted in missense mutations. The proband’s father and grandmother had the same mutation of COL1A1 gene. The mutation was not found in the proband’s mother and normal controls. There was no report for such mutation in Chinese population. Pedigree analysis showed the fami-ly genetic characteristics of autosomal dominant inheritance. The proband was clinically diagnosed as OI type Ⅳwith more severe clinical phenotype. Conclusion PCR-HRM analysis is a new effective method for genetic screening of OI. COL1A1 mutation of c.1138G>A is a newly discovered mutation in Chinese population. Gly replaced inαhelical domain may lead to a more severe clinical phenotype.

OBJECTIVETo detect potential mutations of COL1A1 and COL1A2 genes with polymerase chain reaction-high-resolution melting analysis(PCR-HRMA) in a proband diagnosed with osteogenesis imperfecta (OI).

METHODSPeripheral blood samples were collected from the proband and members of his family as well as healthy controls. The mutations were detected by PCR-HRMA and confirmed by direct sequencing. Potential effects of the mutations were predicted using softwares including PolyPhen, SIFT and Align GVGD.

RESULTSThe PCR-HRMA has indicated mutations in exon 45 of the COL1A1 gene in the proband as well as his parents, which were presented as the difference in the melting curves between the patients and the control samples. Sequencing analysis confirmed that the proband has carried two heterozygous mutations (c.3235G>A, p.Gly1079Ser and c.3247G>A, p.Ala1083Thr) in exon 45 of the COL1A1 gene. Among them, c.3235G>A was predicted to have impeded alpha helix structure domain, which was inherited from the father who also had OI. c.3247G>A was inherited from mother who had a normal phenotype. All three softwares predicted that the c.3235G>A mutation can interfere with the function of the protein, while the c.3247G>A may have a benign effect by PolyPhen analysis.

CONCLUSIONThe study identified two mutations (c.3235G>A and c.3247G>A) occurred simultaneously in COL1A1 gene in a case. The case is the first reported in human collagen mutation database. As identified,mutation of c.3235G>A may be the major cause of the disease in the proband.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Child ; Child, Preschool ; China ; Collagen Type I ; genetics ; Exons ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Osteogenesis Imperfecta ; genetics ; Pedigree ; Point Mutation

OBJECTIVETo investigate mutation of COL1A1 gene and analyze the relationship between genotype and clinical phenotype in a family with osteogenesis imperfecta (OI).

METHODSThe family history of an OI pedigree, along with clinical data, was collected. Blood samples from the proband and his families, as well as 50 normal controls, were collected. Mutation of COL1A1 gene was screened using PCR-high resolution melting (PCR-HRM) and validated by sequencing.

RESULTSPCR HRM method showed an abnormal result in proband COL1A133_34 exons, which Tm was 87.7℃, in contrast to the normal control (wt) Tm of 87.9±0.06℃. There was a significant difference between the proband and the normal control with the standardization curve and the difference curves. DNA sequencing showed that Y9COL1A1 gene exons 33_34 has lost a C base (c.2321delC), which resulted in a frameshift mutation and caused an premature termination codon (UAA) at amino acid 334, i.e., p.Pro774LeufsX334 The father and grandfather of the proband, both suffered from OI, were verified to be heterozygous for the same mutation. The same mutation was not found in 50 normal controls. Database search confirmed this to be a novel mutation. Pedigree analysis suggested that it has an autosomal dominant inheritance. The proband and patients from the family were clinically diagnosed as OI type I.

CONCLUSIONThe study has identified a novel mutation of COL1A1 gene, c.2321delC. This frameshift mutation has caused a premature stop codon and reduced collagen type synthesis, characterized by a lighter OI clinical phenotype.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; Child, Preschool ; China ; Collagen Type I ; genetics ; Female ; Frameshift Mutation ; Humans ; Male ; Molecular Sequence Data ; Osteogenesis Imperfecta ; genetics ; Pedigree