Genotype of thalassemia genes and the polymorphism of β- globin gene in Cantonese / 中华血液学杂志

<p><b>OBJECTIVE</b>To understand the genotype of α and β-globin, as well as the polymorphism of β-globin gene in Cantonese in recent years, and to provide an effective genetic diagnosis for thalassemia (thal).</p><p><b>METHODS</b>The single-tube complex PCR was used to detect 3 types of deletional α-thal, reverse dot blotting (RDB)/PCR to detect 3 kinds of undeletional α-thal-αCS, αQS, αWSand 18 kinds of β-thal mutations which were common in Chinese population. A total of 454 cases from Guangdong were undergone thal genotype genetic diagnosis. Among the 454 cases, 142 cases were selected to perform the single nucleotide polymorphisms (SNPs) analysis of β- globin gene by denaturing high-performance liquid chromatography (DHPLC)combining the whole gene sequencing.</p><p><b>RESULTS</b>Of the 454 cases, 438 were diagnosed as thalassemia, including 246 of α-thal, 164 of β-thal and 28 of αβ-thal. In 246 α-thal cases, deletions were the dominant mutations, including 197 cases of αα/--(SEA), 20 of αα/-α(3.7) and 9 of αα/-α(4.2). In 164 β- thal cases, heterozygotes accounted for 92.7% (152/164), the main genotypes were CD41- 42, IVS-II-654, ﹣28 and CD17, and the dual heterozygotes and homozygotes accounted for 4.9% (8/164) and 2.4% (4/164), respectively. The result of β-globin gene screening by DHPLC combining with sequencing was consistent with that of RDB. Moreover, we also found 9 kinds of SNP, in which 2 were unreported, the IVS-I-13 G> A and IVS-II- 310 T>C. In the tested samples, the frequency of 4 kinds SNP was high, among which 3 kinds SNPs-rs713040, rs10768683 and rs1609812 were carried together.</p><p><b>CONCLUSION</b>The dominant genotypes were αα/--(SEA) in α-thal cases, CD41-42, IVS-II-654, -28 and CD17 in β-thal. The frequency of β-thal heterozygotes, homozygotes and αβ-thal is high. DHPLC combining the whole β-globin gene sequencing can effectively detect the common β-thal mutation and even new mutations or SNPs. In Cantonese, the frequency of SNP rs713040, rs10768683, rs7480526 and rs1609812 of β-globin gene was high, and there may exist genetic linkage between rs713040, rs10768683 and rs1609812.</p>

High expression of nucleophosmin/B23 in hepatocellular carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the expression of nucleophosmin/B23 (B23) in tumor cells of hepatocellular carcinoma (HCC) and its clinicopathologic significance.</p><p><b>METHODS</b>Mouse monoclonal antibodies against B23 were raised by recombinant protein and hybridoma technology. Immunohistochemical study for B23 was performed on 103 cases of HCC, 12 cases of focal nodular hyperplasia and 17 cases of native liver tissue adjacent to hepatic hemangioma. Fresh specimens from 10 cases of HCC and the adjacent liver tissue were also collected for reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of B23 was analyzed and compared with that of proliferative cell nuclear antigen (PCNA) in these specimens.</p><p><b>RESULTS</b>RT-PCR and Western blot analysis showed that B23 expression in HCC was higher than that in adjacent liver tissue. Statistically significant difference in expressions of B23 and PCNA were also noted in the four groups studied (P < 0.01). B23 and PCNA expressions in HCC were higher than those in the other three groups (P < 0.01). There was also a statistically significant correlation between B23 and PCNA expressions amongst the four groups (r = 0.4769, P < 0.01). Besides, B23 expression in HCC correlated with pathologic tumor grading, serum alpha-fetal protein levels and cirrhotic status (P < 0.05).</p><p><b>CONCLUSIONS</b>B23 expression in HCC was significantly higher than that in liver tissues with non-malignant diseases. B23 may be used as a marker for neoplastic changes in liver cells and thus has potential clinicopathologic application.</p>

Identification of G6PD gene variants from Hakka population in Guangdong province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Studying on G6PD polymorphism from Hakka population in Guangdong province.</p><p><b>METHODS</b>Identifying the variants of G6PD gene and determining the frequencies respectively with the use of amplified refractory mutation system(ARMS), polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) and ABI 3100 DNA sequencing technologies.</p><p><b>RESULTS</b>Mutations of G6PD gene in cDNA 1388 (G-->A), 1376 (G-->T), 95 (A-->G), 392 (G-->T), 1024 (C-->T), 1311 (C-->T) have been found.</p><p><b>CONCLUSION</b>G6PD cDNA 1388 (G-->A), 1376 (G-->T), 95(A--> G), 392 (G-->T), 1024 (C-->T) and 1311 (C-->T) accompanied with intron 11 (93 T-->C) are the common mutations in Chinese population. cDNA 1388 (G-->A), cDNA 1376 (G-->T) are the most popular G6PD gene variants in Hakka population. In this study, no new type of G6PD gene mutation was found in the Hakkas of Guangdong.</p>

Complex mutations of 1311 C-->T in exon 11 and 93 T-->C in intron 11 in G6PD gene / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between complex 1311 mutation of C-->T in exon 11 and 93 T-->C in intron 11 of G6PD gene and the G6PD deficiency.</p><p><b>METHODS</b>Using NBT paper strip method to screen and quantitative NBT method to confirm G6PD deficiency. PCR-SSCP technique was used to find the abnormal exon 11 and the amplification refractory mutation system (ARMS) to identify 1311 mutation, and DNA sequencing to identify the complex mutation at 1311 in exon 11 and 93 in intron 11.</p><p><b>RESULTS</b>Abnormal band in exon 11 was found in 12 cases. DNA sequencing showed that they were 1311 mutation together with 93 mutation.</p><p><b>CONCLUSION</b>This complex mutation may be the cause of reduced activity of G6PD enzyme.</p>