Polymorphisms of DNA methyltransferases and the risk of prostate cancer / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the polymorphisms of DNA methyltransferase (DNMT) and the risk and pathologic characteristics of prostate cancer (PCa) in Chinese men.</p><p><b>METHODS</b>This case-control study included 155 PCa patients and 155 healthy male controls. Using Sequenom MassARRAY, we detected the genotypes of the DNMT1 polymorphisms rs16999593 and rs2228611 and the DNMT3B polymorphism rs2424908, followed by analysis of their association with the risk and pathologic characteristics of prostate cancer by logistic regression.</p><p><b>RESULTS</b>Significant differences were found in the frequency of the rs16999593 genotypes (P = 0.041) and that of the rs2424908 genotypes (P = 0.025) between the case and control groups. The frequencies of the genotypes rs16999593CT (OR = 0.61, 95% CI 0.38-0.99, P = 0.043) and rs16999593CT/CC (OR = 0.59, 95% CI 0.39-0.92, P = 0.017) were obviously higher in the control than in the case group, and so were those of rs2424908CT (OR = 0.73, 95% CI 0.58-0.91, P = 0.007) and rs2424908CT/CC (OR = 0.57, 95% CI 0.36-0.94, P = 0.023). The frequencies of rs16999593CT/CC (OR = 0.47, 95% CI 0.28-0.85, P = 0.008) and rs2424908CT/CC (OR = 0.46, 95% CI 0.28-0.85, P = 0.009) were evidently lower in the cases with Gleason score < 7 than in the controls. However, none of the three polymorphisms ex hibited any significant differences in the frequencies of their genotypes between the patients with Gleason score > 7 and the healthy con trols (P > 0.05).</p><p><b>CONCLUSION</b>The rs16999593CT/CC genotype of DNMT1 and the rs2424908CT/CC genotype of DNMT3B are as sociated with decreased risk of prostate cancer and lower Gleason score in C.</p>

Changes of regulatory T cells and T helper cells in peripheral blood and their roles in the severity evaluation in children with asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the changes of CD4+CD25+Foxp3+ regulatory T cells (Treg) and T helper cells (Th1/Th2) in peripheral blood and their roles in the severity evaluation in children with asthma.</p><p><b>METHODS</b>One hundred and fifty children with asthma were classified into acute attack (94 cases) and remission (56 cases) groups according to their clinical features, and the acute attack children were subdivided into mild asthma (54 cases) and severe asthma (40 cases) groups. Fifty healthy children were enrolled as a control group. The levels of CD4+CD25+Foxp3+ Treg, CD4+IFN-γ+ Th1 and CD4+IL-4+ Th2 in peripheral blood were measured by flow cytometer.</p><p><b>RESULTS</b>The mean levels of CD4+CD25+Foxp3+ Treg and the ratio of Th1/Th2 in asthmatic children were lower than those in the control group (P<0.01). The Treg levels and the ratio of Th1/Th2 in the acute attack group were lower than those in the remission group and in the control group (P<0.01). The Treg levels in the severe asthma group were lower than those in the mild asthma group (P<0.01). There was a remarkably negative correlation between Treg levels and the asthma severity (r=-0.737, P<0.01), and the Th1/Th2 ratio was also negatively correlated with the asthma severity (r=-0.615, P<0.01). The Treg levels were positively correlated with the Th1/Th2 ratio (r=0.856, P<0.01).</p><p><b>CONCLUSIONS</b>The Treg levels decrease remarkably and Th subsets imbalance occurs in children with asthma. This suggests that Treg and Th immunity play important roles in the pathogenesis of asthma. The Treg levels and the ratio of Th1/Th2 in peripheral blood may be useful in the evaluation of severity in children with asthma.</p>

Effect of recombinant adenovirus Ad-DT-A in targeted therapy for malignant cancer cell lines with loss of IGF2 imprinting / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the feasibility of IGF2 imprinting system in target gene therapy for tumors.</p><p><b>METHODS</b>The mouse H19 enhancer, DMD and promoter H19 were amplified by PCR from mouse genomic DNA and then cloned into the plasmid pDC312. The EGFP and DT-A fragments were amplified by PCR and cloned into the recombinant plasmid, and then the shuttle plasmid were transfected into HEK293 cells together with the adenoviral vector Ad5, namely, Ad-EGFP and Ad-DT-A. Adenovirus hexon gene expression was applied to confirm the presence of adenovirus infections. The effect of the IGF2 imprinting system was tested by fluorescence microscopy. RT-PCR and Western blotting after transfection of the recombinant adenoviral vectors into cancer cells were used to show loss of IGF2 imprinting (LOI) and maintenance of IGF2 imprinting (MOI), respectively. The anti-tumor effect was assessed by MTT and flow cytometry after the HCT-8 (LOI). Human breast cancer cell line MCF-7 (MOI) and human normal gastric epithelial GES-1 (MOI) cell line were transfected with Ad-DT-A in vitro. The anti-tumor effect was detected by injecting the Ad-DT-A in nude mice carrying HCT-8 tumors.</p><p><b>RESULTS</b>The expression of EGFP protein, DT-A mRNA and DT-A protein were seen to be positive only in the HCT-8 tumor cell line. Infection with Ad-DT-A resulted in obviously growth inhibition in HCT-8 cells (75.4 ± 6.4)% compared with that in the control group, and increased the percentage of apoptosis in the HCT-8 cells (20.8 ± 5.9)%. The anti-tumor effect was further confirmed by injecting the recombinant adenoviruses in HCT-8 tumor-bearing nude mice, and the results showed that the Ad-DT-A inhibited the tumor growth, with on inhibition rate of 36.4%.</p><p><b>CONCLUSIONS</b>The recombinant adenoviruses carrying IGF2 imprinting system and DT-A gene have been successfully constructed, while Ad-DT-A can effectively kill the tumor cells showing loss of IGF2 imprinting. It might play an important role in future target gene therapy against malignant tumors based on loss of IGF2 imprinting.</p>

Study on the relationship between polymorphism sites of IL-10 and the susceptibility of endometriosis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the relationship of the endometriosis susceptibility and polymorphism of up stream of IL-10 promoter at the site of 1082(G→A), 819(C→T) and 592(C→A).</p><p><b>METHODS</b>A total of 214 patients with endometriosis and 160 healthy individuals were enrolled and divided into patient group and control group in this study. The polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) was applied to detect the base transition in the up stream of IL-10 promoter at the site of 1082(G→A), 819(C→T) and 592(C→A). SPSS11.0 software was applied to analysis frequencies of all genotypes.</p><p><b>RESULTS</b>There was no difference in polymorphism of IL-10-1082 between the endometriosis (AA: 87.90%, GA: 12.10%) and control group (AA: 87.50%, GA: 12.50%). The rate of TT, CT and CC genotype IL-10-819 was the same as the AA, CA and CC individually. There was no difference in the polymorphism of IL-10-819 or IL-10-592 between the endometriosis group (TT or AA: 41.12%, CT or CA: 47.66%, CC: 11.21%) and control group (χ(2) = 5.87, P = 0.053). However, there were significant difference in the genotype of CT of IL-10-819 or CA of IL-10-592 between the endometriosis group and control group (after adjust OR = 1.88, 95% CI = 1.10 - 3.21, χ(2) = 5.24, P = 0.021), and the allele C of IL-10-819 or IL-10-592 were close related with occurrence of endometriosis (OR = 1.42, 95%CI = 1.04 - 1.95, χ(2) = 4.81, P = 0.028). The IL-10 level in the plasma of endometriosis group with genotype of CC (CC), CT (CA) of IL-10-819(-592) were significant higher than those with TT (AA) (CA/CT: (50.12 ± 82.40) pg/ml, CC: (91.00 ± 118.23) pg/ml, TT/AA: (21.45 ± 22.10) pg/ml) (F = 2.492, P = 0.048; F = 1.852, P = 0.008).</p><p><b>CONCLUSION</b>The allele C of IL-10-819 or IL-10-592 was close related to the high level expression of IL-10, and it is the risk of the occurrence of endometriosis.</p>

Influence of SNCG transfection on the effectiveness of anti-tumor drugs in PC-3 cell lines / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the sensitivity of the PC-3 cell lines transfected with the PCI-NEO-SNCG plasmid to Cisplatin (DDP), 5-Fluorouracil (5-FU), Adriamycin (ADM), Vincristine (VCR) and Paclitaxel (TAX), and to explore the influence of the SNCG expression on the effectiveness of anti-tumor drugs.</p><p><b>METHODS</b>The plasmids PCI-NEO and PCI-NEO-SNCG were transfected into the hormone-independent prostate cancer cell lines PC-3. RT-PCR was adopted to examine the expression of SNCG in the PC-3 cell lines. The MTT method was employed to detect the suppressive effects of different anti-tumor drugs (DDP, ADM, 5-FU, VCR and TAX) on the cell lines transfected with PCI-NEO and PCI-NEO-SNCG. Flow cytometry was used to analyze the cell cycles and apoptosis of the transfected cells treated with TAX.</p><p><b>RESULTS</b>The 5 anti-tumor drugs suppressed the growth of the cell lines transfected with the plasmids PCI-NEO and PCI-NEO-SNCG in a time-dependant manner. The comparison between the growth-suppressing effects of different anti-tumor drugs on the PC-3 cell lines showed no significant differences between the group transfected with PCI-NEO and that with PCI-NEO-SNCG in DDP, 5-FU, ADM and VCR (P > 0.05), while the rate of suppression of TAX on the latter cell lines was significantly lower than that on the former (P < 0.01). Compared with the PCI-NEO-SNCG plasmid transfected cell lines, after treated with TAX for 48 hours, those transfected with the PCI-NEO plasmid exhibited a significantly larger proportion of cells remaining in the G2-M stage (P < 0.01), a smaller proportion in the G0-G1 and S stages (P < 0.01) and a significantly higher expression of Caspase-3 (P < 0.01).</p><p><b>CONCLUSION</b>The significant reduction of the growth-suppressing effect of TAX in the SNCG-transfected PC-3 cell lines suggests that the expression of SNCG may restrain the effect of TAX. These findings have provided evidence and guide to the individual chemotherapy of prostate cancer.</p>