Methylenetetrahydrofolate Reductase Gene Polymorphism C677T is Associated with Increased Risk of Coronary Heart Disease in Chinese Type 2 Diabetic Patients / 中国医学科学杂志(英文版)

Objective Chronic cardiovascular diseases induced by long-term poor blood glucose control are the main cause of death in patients with type 2 diabetes mellitus (T2DM). Previous researches report that methylenetetrahydrofolate reductase gene (

Diagnostic Value and Clinical Significance of G6PD Activity for the Mediterranean Anemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the diagnostic value and clinical significance of the glucose 6-phosphate dehy-drogenase (G6PD) activity for mediterranean anemia (MA), so as to provide the reference for early clinical diagnosis and treatment of MA.</p><p><b>METHODS</b>The peripheral blood was collected from 100 healthy persons and 168 patients with MA, then the agarose gel electrophoresis, MA gene detection, blood routine examination, serum ferrium levels and G6PD activity assay were performed, and the results of evaluating MA were comparatively analyzed.</p><p><b>RESULTS</b>The G6PD activity in all type MA patients was obviously higher than that in healthy controls (P < 0.01), the MCV value in all type MA patients was significantly lower than that in healthy controls (P < 0.01). The detection of G6PD activity showed that the sensitivity, specificity, positive and negative likelihood ratio, diagnostic index and Youden index of G6PD for MA patients were 85.12%, 68%, 2.66, 0.219, 1.53 and 0.53 respectively, which suggest the better efficacy of G6PD value for diagnosis of MA.</p><p><b>CONCLUSION</b>The G6PDS activity of patients with MA in different subtypes is higher than that of healthy persons, the G6PD level has a certain diagnostic value for MA, but there is an optimal range.</p>

Bufo gargarizans mcl-1 cloning and its prokaryotic recombinant protein expression / 药学学报

MCL-1 is encoded by myeloid cell leukemia-1 gene (mcl-1), which is one of the anti-apoptotic members of bcl-2 cell apoptotic gene superfamily. ChanSu is made of dorsal secretions of several Bufo species, commonly used in the prescriptions of traditional Chinese medicine for treating many diseases including cancer. To clarify if mcl-1 is expressed in the dorsal skin of B. gargarizans, the PCR (polymerase chain reaction) was performed with its dorsal skin first strand cDNA as the template and a pair of specific primers of mcl-1, and PCR products were cloned into the pGM-T vector. DNA sequencing indicated that the ORF length was 639 bp encoding 212 amino acid residues, and the homology of 44%-95% with the MCL-1 of several other animals. For the further studies on MCL-1 biological functions during the oncogenesis and preparation of its antibody, the prokaryotic expression construct of pET-28b-mcl-1 was prepared which was confirmed by DNA sequencing, and its recombinant protein expression (0.02% wet weight) in E. coli BL21 (DE3) strain was confirmed by SDS-PAGE and Western blotting.

Cloning and bioinformatic analysis of TAGLN2 cDNA of Bufo japonicus formosus / 药学学报

To study the bioactive polypeptides included in Bufo skin and its secretions the plasmid skin cDNA library of adult Japanese toad Bufo japonicus formosus was prepared. The pSD64TR has been used as the vector and the cloning sites are Xho I and EcoR I. To screen cDNAs encoding bioactive components, the plasmid cDNA library was transformed into E. coli DH5 competent cells, and positive colonies were screened by colony PCR (polymerase chain reaction). The suspension of a single colony in LB medium was used as the template, SP6 (the upstream primer of the plasmid cDNA library) and a primer with Xho I site and polyT were used as the primers. As the result, 465 positive colonies out of 1 344 were obtained and their plasmid were collected and sequenced. By homologous analysis, it was found that one of the cDNAs encoding a peptide with high homolog with transgelin-2, which was registered in GenBank (accession number: JX197456), and it was indicated as a partial cDNA sequence with a deletion at the 5' end. The transcript is 997 bp consisting of 31 bp 5', 618 bp 3' untranslated region (UTR) and an open reading frame (ORF) of 348 bp encoding a polypeptide of 115 amino acids. In the putative protein product, there is a calponin homology domain, two cysteine residues for a disulfide bond and three a-helix domains, and five potential phosphorylation sites. The homologous analysis indicates 90% similarity with Xenopus (Silurana) tropicalis and 89% with Xenopus laevis, and 71%-85% with other species.