Apolipoprotein M expression in human colorectal cancer tissues and its clinicopathological relevance / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the mRNA and protein expression levels of apolipoprotein M (apoM) in the human colorectal cancer tissues, and to explore its clinical relevance.</p><p><b>METHODS</b>Real-time PCR was carried out to determine the mRNA expression levels both in cancer tissue and its adjacent normal tissue from 20 patients with colorectal cancer. Immunohistochemistry was also carried out to determine the protein levels in 23 colorectal biopsy samples (7 normal mucosa, 6 inflammatory mucosa and 10 polyp tissues) and 20 cases of colorectal cancer tissues as well as the adjacent normal tissues.</p><p><b>RESULTS</b>Real-time PCR result showed that apoM mRNA level in the colorectal cancer tissues was significantly lower than that in their adjacent normal tissues (0.05±0.01 vs. 0.19±0.05, P<0.05). ApoM mRNA level in colorectal cancer tissues was statistically significant higher in the patients with lymph node metastasis as compared to the patients without lymph node metastasis (P<0.01). The median value of apoM protein in cancer tissues was 5.50, which was significantly lower than that in the adjacent normal tissues (10.5, P<0.05), inflammatory mucosa tissues (9.75, P<0.05), polyp tissues (11.0, P<0.01) and normal mucosa (10.5, P<0.05). No significant association was observed between the apoM protein level and the clinicopathological parameters of patients.</p><p><b>CONCLUSIONS</b>Both apoM mRNA and protein expression levels in colorectal cancer tissues are significantly decreased in contrast to normal and benign colorectal tissues. The apoM mRNA expression in colorectal cancer tissues is closely associated with nodal metastasis.</p>

A novel method of detecting mitochondrial C1494T and A1555G mutations by using the base-quenched probe technique in a single PCR assay / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>In the present study we describe a new method to detect the mitochondrial DNA (mtDNA) mutations A1555G and C1494T by using the base-quenched probe technique in a single PCR reaction.</p><p><b>METHODS</b>6-carboxyfluorescein (FAM) was directly conjugated to the 3' end of the probe. Four vectors, representing the four possible genotype combinations, were constructed as the amplification template for the methodology established. In present study A1555G and C1494T mutations in 117 individuals with hearing loss were detected by the base-quenched probe method and were further validated by the direct DNA sequencing analyses.</p><p><b>RESULTS</b>From the melting curve we could distinguish the four haplotypes accurately. And there were complete concordance between the base-quenched probe method and direct DNA sequencing.</p><p><b>CONCLUSION</b>This method is suitable for clinical test of mtDNA mutations A1555G and C1494T in individuals with hearing loss.</p>

Alterations of CD4+ CD25+ regulatory T cells in patients with idiopathic thrombocytopenic purpura / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the role of CD4+ CD25+ regulatory T (Treg) cells in patients with ITP.</p><p><b>METHODS</b>Flow cytometry was used to examine the number of CD4+ CD25+, CD4+ CD25 high, CD4+ Foxp3+ and CD4+ CD25+ Foxp3+ T cells. The level of Foxp3 mRNA expression was analyzed by realtime quantitative reverse transcriptase polymerase chain reaction (RT-PCR) . CD4+ CD25 high T cells was cocultured with CD4+ CD25 - T cells from patients or controls for assessing the regulatory properties of CD4+ CD25 Treg cells.</p><p><b>RESULTS</b>The proportion of CD4+ CD25+ T cells in the peripheral blood of patients with ITP [(15.64 +/- 5.82) %] was significantly higher than that in normal control group [(9.30 +/- 3.95)%] (P <0.01). There was no significant difference in the percentages of CD4+ CD25 high T cells between ITP patients and controls [(1.53 +/- 0.66)% versus (1.36 +/- 0.55)% (P = 0.317)]. But the number of CD4 Foxp 3+ T cells and CD4 + CD25+ Foxp3+ T cells in patients were significantly lower than that in control group (P <0.01). The expression of Foxp3 mRNA reduced (P < 0.01) and the suppressive activity of CD4+ CD25 high T cells is lower than that of healthy controls (P <0.01).</p><p><b>CONCLUSION</b>In patients with ITP, both the number and immuno-regulative function of CD4+ CD25+ Treg cells are reduced.</p>

Expression and clinical significance of costimulatory molecule B7-H3 mRNA and B7-H3 protein in gastric carcinoma / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study the expression of B7-H3 mRNA and B7-H3 protein in gastric carcinoma and their clinical significance.</p><p><b>METHODS</b>The expression of B7-H3 mRNA and B7-H3 protein in gastric carcinoma and the nearby normal tissue of 38 patients was detected by real-time RT-PCR and immunohistochemical assay respectively.</p><p><b>RESULTS</b>B7-H3 mRNA was expressed both in gastric carcinoma and nearby normal tissue, but the expression level in gastric carcinoma was much lower than that in nearby normal tissue. There were no significant differences of B7-H3 mRNA expression among gender, age, histological type, tumor size, lymph node metastasis and invasive depth (all P >0.05). The positive rate of B7-H3 protein expressed in gastric carcinoma was 39.5%. There were no significant differences of B7-H3 protein expression among gender, age, histological type, tumor size, lymph node metastasis and invasive depth (all P >0.05), but there were significant differences among groups of clinical stage (P=0.022) and pathological grade (P=0.039). Kaplan-Meier analysis revealed that disease-free survival or overall survival of the patients with positive B7-H3 expression were significantly longer than those with negative B7-H3 expression (P=0.009 and P=0.010 respectively).</p><p><b>CONCLUSION</b>Detection of B7-H3 expression in gastric carcinoma will be beneficial to the judgment of the prognosis of gastric carcinoma and the choice of individualized treatment.</p>

Relationship between DNA degradation and postmortem interval of corrupt corpse / 法医学杂志

OBJECTIVE@#To study the relationship between DNA degradation and postmortem interval of corrupt corpse.@*METHODS@#By determining the marrow DNA content with histochemical technique and image analysis.@*RESULTS@#The content of marrow DNA decreased gradually with prolongation of postmortem interval, and it evencould be detected till 14 days after death.@*CONCLUSION@#There was a linear relationship between the degradation rate of the nuclear DNA and postmortem interval.

A novel method for detecting single-nucleotide polymorphisms by using ShineRoar probes / 中华检验医学杂志

Objective The present study demonstrates a novel,simple and cost-effective method for detecting known SNP genotyping by using ShineRoar probes.Methods The SNP of target genes detected by using the ShineRoar probes and melting curve analysis.Tumor necrosis factor receptor Ⅱ (TNFR Ⅱ) and apolipoprotein M (apoM) had been employed as target genes to describe the method in details.The PCR products of TNFR Ⅱ and apoM were collected and sequenced.Results The melting temperatures (TM) were significantly different between mutated genotypes and wild-type genotype.A biallelic SNP marker (T/ G) at position 196 in exon 6 of TNFR Ⅱ gene showed two melting valleys with the appropriate TMs at (52.84?0.75)℃ and (58.38?0.61)℃,respectively.For apoM T-778C,TMs of homozygous T genotype and C genotype were (42.55?0.73)℃ and (49.19?0.57)℃,respectively.Moreover,this genotyping method was validated by the DNA sequence analyses (Kappa=1,P=0.000).Conclusion It is concluded that this novel method is simple and economical and it is suitable for a large-scale genotyping screening.