Effect of electroacupuncture on pain threshold and spinal NR2B subunit expression in a rat model of neuropathic pain / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of electroacupuncture (EA) on pain threshold and spinal NR2B subunit expression in a rat model of neuropathic pain due to chronic compression injury (CCI) of the sciatic nerve and explore the analgesic mechanism of EA.</p><p><b>METHODS</b>Male SD rats weighing 200-280 g were randomly divided into 4 groups (n=10), namely the sham-operated group, CCI group, EA+CCI group, and sham EA+CCI group. All the rats underwent tests of the mechanical withdrawal threshold and thermal threshold. On day 13 after the surgery, all the rats were decapitated to collect the L4-6 segments of the spinal cord to examine NR2B expression using Western blotting.</p><p><b>RESULTS</b>The postoperative mechanical withdrawal threshold and thermal threshold were significantly lowered in CCI, EA+CCI and sham EA+CCI groups as compared to those before the surgery (P<0.05). EA obviously alleviated the hypersensitivity in the rats with CCI and inhibited spinal NR2B expressions (P<0.05). No significant differences were found in the mechanical withdrawal threshold, thermal threshold or spinal NR2B subunit expression between CCI group and sham EA+CCI group (P>0.05).</p><p><b>CONCLUSION</b>EA may alleviate neuropathic hypersensitivity partially by inhibiting NR2B expression in the spinal cord of rats with neuropathic pain resulting from CCI of the sciatic nerve.</p>

Construction of an eukaryotic expression vector for PRL-2 and its effect on human hepatocellular carcinoma cell invasiveness and migration in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct an eukaryotic expression vector for PRL-2 and evaluate its effect on the invasiveness and migration of a human hepatocellular carcinoma cell line.</p><p><b>METHODS</b>RT-PCR was performed to amplify the complete PRL-2 open reading frame using the total mRNA of hepatocellular carcinoma HepG2 cells as the template. PRL-2 gene was inserted into the pGEM T easy vector and sequenced, and the correct PRL-2 sequence was subcloned into the mammalian expression vector pcDNA3.1+. The constructed PRL-2 vector was transfected into CL1 cells via lipofectamine, and the stable expression of PRL-2 mRNA was detected by RT-PCR, the expressed protein identified by immunohistochemistry and Western blotting, and the effect of PRL-2 on the adhesion ability of CL-1 cell evaluated with MTT assay 20 and 120 min after transfection. The effect of PRL-2 on the invasive migration of CL-2 cells was evaluated according to the number of cells penetrating the Matrigel layer of polycarbonate membrane of Boyden chamber.</p><p><b>RESULTS</b>RT-PCR yielded a fragment of 504 bp and the inserted PRL-2 sequence was verified by sequence analysis. The subclones were identified by restriction endonuclease digestion, and a G418-resistant clone, PRL-2-CL1, was obtained after 8 weeks of selection. RT-PCR showed stable expression of PRL-2 mRNA, and Western blotting confirmed overexpression of PRL-2 protein in the transfected cells. PRL-2 increased the adhesion rate of CL-1 cells to fibronectin at 20 min and 120 min after transfection (P<0.05), and also the number of CL-1 cells penetrating the polycarbonate membrane from 10.0+/-3.7 to 44.8+/-2.6 (P<0.05).</p><p><b>CONCLUSION</b>An eukaryotic expression vector of PRL-2 has been successfully constructed, which allows stable and efficient expression in CL-1 cell line. PRL-2 can promote cell adhesion and invasion activity of human hepatocellular carcinoma cells.</p>

Association of XAGE-1b gene expression with clinical characteristics of non-small cell lung cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the association between XAGE-1b gene expression and the clinical characteristics of non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Tumor tissue and adjacent normal lung tissue specimens were obtained surgically from 30 patients with resectable NSCLC, from which the total RNA was extracted for RT-PCR to amplify full-length XAGE-1b gene. The products of RT-PCR were identified by electrophoresis and sequencing. The expression of XAGE-1b gene and its association with the clinical characteristics of the patients were analyzed.</p><p><b>RESULTS</b>In the 30 tumor tissue specimens, the expression rate of XAGE-1b gene was 40%, but none of the normal lung tissues expressed this gene. The gene expression was not related to the patients' age, gender, tumor differentiation or clinical stages, but showed significant correlation to their pathological classification. The expression rate of XAGE-1b gene in adenocarcinoma was much higher than that in tumors of other pathological types (61.1% vs 8.3%, P=0.015). XAGE-1b gene expression tended to increase with the TNM stages, which, however, failed to find statistical data support (P>0.05).</p><p><b>CONCLUSIONS</b>XAGE-1b gene is highly expressed in lung adenocarcinoma, and can be an ideal target for tumor immunotherapy.</p>

Expression of NY-ESO-1 and LAGE-1 cancer-testis antigens in hepatocellular carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the role of NY-ESO-1 and LAGE-1 cancer-testis antigens as targets for immunotherapy and the relationship between corresponding gene expression and biologic behavior of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The expression of NY-ESO-1 and LAGE-1 was studied in frozen tumor tissues from 30 cases of HCC by reverse transcriptase-polymerase chain reaction and immunohistochemistry. NY-ESO-1 expression and its distribution were further studied by immunohistochemistry in a tissue array contained 191 cases of HCC.</p><p><b>RESULTS</b>NY-ESO-1 and LAGE-1 mRNAs were expressed in 33.3% (10/30) and 16.7% (5/30) of HCC respectively. Either NY-ESO-1 or LAGE-1 was expressed in 36.7% (11/30) cases. NY-ESO-1 was expressed mainly in the cytoplasm of tumor cells. It was positive in 13.8% (24/174) cases of HCC. There was an increased expression of NY-ESO-1 from 6.8%, 3/44 in small HCC, 16.2%, 21/130 in advanced HCC and 23.1%, 12/52 in metastatic HCC. The expression in the non-metastatic group was 9.8% (12/122). The differences between the metastatic group and non-metastatic group (< 0.05) and between normal liver tissue and HCC (< 0.01) were statistically significant. There was no relationship between NY-ESO-1 expression and tumor size. NY-ESO-1 and LAGE-1 were not detected in adjacent normal liver tissue.</p><p><b>CONCLUSIONS</b>NY-ESO-1 and LAGE-1 are expressed in a high percentage of HCC, especially in cases with metastasis. It is thus possible that NY-ESO-1/LAGE-1 can serve as targets for antigen-specific immunotherapy in HCC and NY-ESO-1 peptide vaccination may be of use for patients with advanced HCC.</p>