Phenotype Types and Genetic Mutation Mechanism of Rhesus D Variant Individuals / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the phenotype types and genetic mutation mechanism of Rhesus D variant individuals.</p><p><b>METHODS</b>Fouty-eight peripheral blood samples of pregnancies and blood donors who had been identified as Rhesus D variant by using routine serologic methods were collected from January 2013 to October 2015 in our center. The multiple ligation-dependent probe amplification(MLPA) was used to determine the RHD after genomic DNA had been extracted from the blood sample, then the data including gene copy number variations, point mutations, deletions and hybrid fusions were analyzed by GeneMarker software. All exons of blood sample RHD were amplified via PCR and analyzed by sequencing when its MLPA results were not in accordance with serologic results. Cloning and haplotype sequencing were performed if novel allele had been found.</p><p><b>RESULTS</b>Rh phenotypes of the 48 samples were typed as following: 20 cases out of 48 were CcDee(41.7%, 20/48),12 cases were ccDEe (25%,12/48), 11 cases were CCDee(22.9%, 11/48), 5 cases were CcDEe (10.4%, 5/48), respectively. The MLPA analysis showed that 38 cases possessed only 1 variant allele(RHD zygosity was Dd), while 10 cases possessed 2 variant alleles(RHD zygosity was DD). In Dd type individuals, point mutations were found in 18 cases and RHD/CE hybrid fusions were found in 20 cases. In DDindividuals, point mutations combined with RHD/CE hybrid fusions were found in 9 cases, deletion combined with RHD/CE hybrid fusions were found in 1 case. Variant alleles analysis basing on MLPA showed that 14 cases were weak D 15 and 22 cases were RhD VI type 3, however, the variant alleles were not identified in 7 cases due to lack of detecting probes and were identified via sequencing analysis. Two novel mutations, 79-81delCTC and 689G>A were also certificated by sequencing in 2 cases.</p><p><b>CONCLUSION</b>CcDee is the major Rh phenotype in RhD variants, weak D 15 and RhD VI type 3 are the main serologic type of RhD variants, point mutation and RHD/CE hybrid fusions are main molecular mechanism for RhD variant phenotype. Besides, 79-81delCTC and 689G>A are two novel alleles.</p>

Expression of PSF1 in colon cancer tissues and its effect on the proliferation of colon cancer cells / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To detect the expression of PSF1 (partner of Sld five 1) in colon cancer specimens, and to explore the effect of RNA interference targeting PSF1 on the proliferation of colon cancer cells and its mechanism.</p><p><b>METHODS</b>Expression level of PSF1 protein in colon cancer specimens was detected by Western blot in 40 patients with colon cancer from May 2004 to December 2006. The short hairpin RNA (shRNA) plasmid targeting PSF1 was transfected into LOVO, HT-29 and HCT116 cells with liposome, then the expression level of PSF1 protein was measured by Western blot, the effect of PSF1 shRNA plasmid transfection on cell proliferation by MTT assay, anchorage-independent growth by soft agar colomy-formation assay, and PSF2, PSF3 and SLD5 mRNA expression by quantitative reverse transcription polymerase chain reaction.</p><p><b>RESULTS</b>The relative expression level of PSF1 protein in colon cancer tissues was 0.485±0.113, which was significantly higher than that in adjacent normal mucosa tissues (0.056±0.014, P<0.01). Western blot showed that the expression level of PSF1 protein was significantly decreased in colon cancer cells transfected with PSF1 shRNA plasmid. After PSF1 shRNA plasmid transfection, cell proliferation was significantly suppressed, the soft agar colony-forming rates of LOVO, HT-29 and HCT116 cells were significantly lower than those in control groups (P<0.05), meanwhile the expression levels of PSF2, PSF3 and SLD5 mRNA were significantly decreased (P<0.05).</p><p><b>CONCLUSIONS</b>PSF1 is significantly up-regulated in colon cancer tissues compared with adjacent normal mucosa tissues. ShRNA plasmid targeting PSF1 can inhibit the expression of PSF1 gene, suppress the proliferation of colon cancer cells, suggesting that it may be a new therapeutic target for colon cancer.</p>

Expression and clinical significance of GINS complex in colorectal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of GINS complex in colorectal cancer (CRC).</p><p><b>METHODS</b>The expression level of GINS complex including PSF1, PSF2, PSF3 and SLD5 in CRC specimens (n=76) were detected by real-time fluorescent quantitative polymerase chain reaction. The association of GINS complex with clinicopathological parameters and prognosis of CRC patients were analyzed.</p><p><b>RESULTS</b>The relative expression level of PSF1, PSF2, PSF3, and SLD5 mRNA in CRC tissues was 0.001 853±0.000 651, 0.007 757±0.004 260, 0.000 967±0.000 481 and 0.003 248±0.001 721, which was significantly higher than that in normal colorectal mucosa tissues (0.000 352±0.000 169, 0.002 951±0.001 216, 0.000 472±0.000 271, and 0.001 675±0.001 156) (all P<0.01). PSF1 mRNA expression was associated with tumor size (P<0.01), and PSF2 mRNA expression with age (P<0.05) and lymph node metastasis (P<0.05). No correlations between PSF3 mRNA expression and clinicopathological parameters were observed. SLD5 mRNA expression was associated with lymph node metastasis (P<0.01). Patients with high expression of PSF1, PSF2 and SLD5 had worse 5-year overall survival rate (57.1%, 54.3%, and 54.3%) than those with low expression (77.1%, 80.0%, and 80.0%) (all P<0.05). Multivariable Cox regression analysis indicated that PSF1 mRNA expression (P<0.05) was an independent factor associated with prognosis of colorectal cancer.</p><p><b>CONCLUSIONS</b>Overexpression of GINS complex in CRC is associated with clinicopathological characteristics and prognosis of colorectal cancer. PSF1 expression is prognostic for CRC patients.</p>