Expression of a novel human retroviral NP9 gene and potential roles of its protein in systemic lupus erythematosus patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the expression of a novel retroviral (NP9) gene transcripts and the possible role of its protein in systemic lupus erythematosus (SLE) patients.</p><p><b>METHODS</b>The retroviral NP9 gene in SLE patients was isolated and cloned using RT-PCR and TA cloning techniques, and it was analyzed by sequencing. The expression of the NP9 genes in 40 patients with SLE and 48 normal controls using RT-PCR was detected. NCBI BLAST and DNASIS 3.1 software were used to analyze the features of protein of NP9 gene.</p><p><b>RESULTS</b>The positive ratio (77.5%) of the mRNA expression of the retroviral NP9 gene in SLE patients is significantly higher than that (8.3%) in normal subjects (P<0.01). The recombinant NP9 protein comprises 74 AA with pI 9.59. Amino acid sequence analysis indicates that the retroviral NP9 protein shares higher homologies with several human proteins with important biological functions.</p><p><b>CONCLUSION</b>SLE patients possess specific novel retroviral NP9 transcripts. The expression of the retroviral NP9 gene may involve in the genesis or development of SLE.</p>

Preliminary clinical observation on effect of soduim ferulate in treating diabetic nephropathy / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of sodium ferulate (SF) on diabetic nephropathy (DN).</p><p><b>METHODS</b>Forty-eight DN patients of early stage and 54 DN patients of clinical stage were randomly divided into two groups, the conventional treatment group and the SF treatment group. Indexes, including urinary albumin excretion rate (UAER), serum endothelin (ET), blood urea nitrogen (BUN), serum creatinine (Scr) and fasting blood glucose (FBG) were observed.</p><p><b>RESULTS</b>The levels of UAER, BUN and ET were decreased in all DN patients, either early stage or clinical stage, after treated with SF for 4 weeks (P < 0.05, P < 0.01), but changed insignificantly in those treated with conventional treatment.</p><p><b>CONCLUSION</b>SF can decrease the levels of UAER and BUN in DN patients, the mechanism may relate with the decreasing of ET production and antagonizing to the binding of ET with its receptors.</p>

Molecular cloning of a novel retroviral NP9 gene of human endogenous retrovirus and its viral protein expression / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the relationship between retroviruses and autoimmune diseases, to clone the novel retroviral NP9 gene from human endogenous retrovirus (HERV), and to construct its expression vector.</p><p><b>METHODS</b>The viral NP9 gene was amplified and cloned by RT-PCR and T-A clone techniques, and its sequence was determined with Perkin-Elmer 377 DNA Sequencer. The amplified viral NP9 gene was subcloned into the prokaryotic express vector pQE30. The recombinant plasmids were identified by restriction endonuclease digestion and sequencing. The recombinant pQE30-NP9 protein was expressed in M15 host cells under the IPTG induction and showed with SDS-PAGE,and the corresponding NP9 viral protein was identified with Western blot analysis.</p><p><b>RESULT</b>A specific band of 250 bp was amplified using RT-PCR from total RNA of peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE) and confirmed as the NP9 gene via T-A clone and DNA sequencing analyses. SDS-PAGE profile showed a clear protein band with a relative molecular weight 9 kD in the IPTG-induced samples, which was confirmed as viral NP9 protein by Western blot analysis.</p><p><b>CONCLUSION</b>The NP9 gene has been successfully isolated and cloned from PBMCs of SLE patients and the corresponding NP9 viral protein expressed in prokaryotic expression vector.</p>

Study of functional L1 retrotransposon in human type 2 diabetes susceptibility loci / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the susceptibility gene of type 2 diabetes mellitus (T2DM) through a novel strategy.</p><p><b>METHODS</b>Firstly, the common feature of the putative susceptibility genes in the reported susceptibility loci was searched by using NCBI BLAST, and a functional L1 retrotransposon in the loci was found. Secondly, the mRNA expression level of the functional L1 retrotransposon in 25 Han T2DM patients and 22 normal controls was investigated by reverse transcription-polymerase chain reaction, and statistical analysis was implemented in statistical package SPSS10.0. Thirdly, L1 retrotransponson genome mutation screening was performed via sequencing.</p><p><b>RESULTS</b>Screening the human genome for the retrotransposon genome via alignment with the L1 genome using NCBI BLAST showed the functional L1 retrotransposons distribute on most chromosomes except for chromosomes 19, 21 and Y on which rare type 2 diabetes susceptibility loci were reported to reside, and their distribution sites are consistent with the locations of the reported candidate type 2 diabetes susceptibility loci. The mRNA expression level of the functional L1 retrotransposon in the T2DM patients was significantly lower than that in normal subjects (P<0.001). Nonsense mutations including deletion and/or point mutations were observed in all of the 6 T2DM patients tested, but no mutation was observed in all of the 4 normal controls tested.</p><p><b>CONCLUSION</b>The functional L1 retrotransposon may be a candidate susceptibility gene of type 2 diabetes or a key regulator of the susceptibility genes, and it may be an ideal candidate biomarker for screening type 2 diabetes.</p>