ABSTRACT
<p><b>OBJECTIVE</b>To construct an eukaryotic expression plasmid for AY358935 gene and explore its function.</p><p><b>METHODS</b>cDNA of the AY358935 gene was amplified by reverse transcription-PCR and cloned into pGEM-Teasy. The pGEM-T-AY was validated by sequencing and served as a template for the construction of eukaryotic expression plasmid. The pcDNA3.1-AY recombinant was validated by double enzyme digestion and used for transient transfection of M14 cells. Expression of the AY358935 protein and proliferation of the M14 cells were determined respectively by Western blotting and 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) colorimetry.</p><p><b>RESULTS</b>The amplicons of RT-PCR were confirmed to have similar size with the cDNA fragment of the AY358935 gene as well as cloned region of pcDNA3.1-AY. The cloned region of pGEM-T-AY was sequenced to be identical with cDNA sequence of the AY358935 gene. M14 cells were transfected by the AY358935 gene, pcDNA3.1 and liposomes, respectively. After 48 h, expression of the AY358935 protein in M14 cells transfected with the AY358935 gene was significantly higher than other two groups. They also had a significantly higher absorbance value (A=0.74) than other two groups (A=0.39 and 0.46, respectively; P<0.05).</p><p><b>CONCLUSION</b>An eukaryotic expression plasmid of the AY358935 gene was successfully constructed. Product of the AY358935 gene may promote the proliferation of M14 cells.</p>
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the clinical value of contrast-enhanced ultrasonography (CEUS) in assessing the high-risk population of hepatic malignant tumor (HMT).</p><p><b>METHODS</b>Three hundred patients with high-risk of HMT were enrolled and examined by CEUS. The clinical data of these patients were collected. A comparative analysis was performed to assess the ability of CEUS for detecting and characterizing lesions in the liver. Kappa test was used for assessing the intra-and inter-observer reliability of CEUS.</p><p><b>RESULTS</b>In 87 enrolled patients with 119 malignant lesions detected by contrast-enhanced MRI, 112 lesions were detected by CEUS and 95 lesions were detected by unenhanced ultrasonography (US). The detection rate of CEUS for HMT was significant higher than that of US (94.1% vs. 79.8%; P < 0.01). More HMT lesions were detected by CEUS than by US in 17.2% patients. One hundred and seventy-seven patients with 215 lesions were confirmed by pathological diagnosis or long-time follow-up. There were 118 malignant and 97 benign lesions. The accuracy of CEUS for differential diagnosis of the liver lesions was 91.6% (197/215), significantly higher than that by US (59.1%, 127/215) (P < 0.01). CEUS improved the accuracy for 35.0% (62/177) patients. For 96 patients with 105 lesions detected for the first time, the agreement of diagnosis by CEUS was 92.4% (97/105). The reliability of CEUS was high (Kappainter = 0.866; Kappaintra = 0.934).</p><p><b>CONCLUSIONS</b>CEUS improves the detection rate and diagnostic accuracy rate of the HMT high-risk population, with a high agreement for diagnosing the new lesions. CEUS may be considered as a first-line method to assess the high-risk population of HMT conveniently, accurately and reliably.</p>