Effects of inhibited IGF-IR expression on proliferation and apoptosis of human hepatocellular carcinoma cell lines / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic value of inhibiting the expression of insulin-like growth factor-I receptor (IGF-IR) using picropodophyllin (PPP) by studying the effects on proliferative and metastatic potentials of human hepatocellular carcinoma (HCC) using an in vitro cultured cell system.</p><p><b>METHODS</b>IGF-IR expression in human HCC cell lines (Bel-7404, Bel-7402, HepG2, and Huh-7) and human hepatocytes (L02) was assessed at baseline (pre-treatment) and after PPP treatment by western blotting. Changes in cell cycle were analyzed by flow cytometry and in cell viability by sulforhodamine B staining. Early apoptosis was detected by annexin-V/FITC and propidium iodide double-staining assay. Caspase-3/7 activity was suppressed by z-VAD-FMK and analyzed by homogeneous luminescence assay. Effects on cell motility were tested by wound-scratch test. Between-group differences were assessed by t-test or one-way analysis of variance.</p><p><b>RESULTS</b>IGF-IR was markedly up-regulated in all HCC cell lines (vs. non-hepatoma hepatocytes). HCC cells with PPP-inhibited IGF-IR showed time-dependent decreases in cell motility and viability. After treatment with PPP for 24 hours, the proportion of HCC cells in G1 phase was 2.1% +/- 0.4%, in S phase was 11.0% +/- 0.7%, and in G2/M phase was 87.1% +/- 0.6%, and no healing was observed in the wound-scratch assay. The PPP treatment induced cell apoptosis, as evidenced by enhanced caspase-3/7 activity; the proportion of annexin-V+/PI- cells was significantly higher in the HepG2 cells than in the non-hepatoma hepatocytes (16.4% +/- 0.4% vs. 5.8% +/- 0.2%, t = 14.05, P less than 0.01). After z-VAD-FMK treatment, the apoptosis rate was significantly higher in the HepG2 cells than in the non-hepatoma hepatocytes (11.3% +/- 0.7% vs. 5.8% +/- 0.2%, t = 11.83, P less than 0.01).</p><p><b>CONCLUSION</b>IGF-IR is associated with proliferation, cell motility, and apoptosis of HCC cells, and may be a promising molecular target for HCC.</p>

Correlation between epigenetic alterations in the insulin growth factor-II gene and hepatocellular carcinoma / 中华肝脏病杂志

To investigate whether epigenetic alterations in the insulin-like growth factor-II (IGF-II) gene that cause differential transcription or expression are correlated with onset and severity of hepatocellular carcinoma (HCC). Patient-matched specimens of HCC, paracancerous, and non-cancerous tissues were collected from 40 primary liver cancer patients. Epigenetic alterations in the promoter (P3) sequence of the IGF-II gene were analyzed by methylation-specific PCR (MSP) and IGF-II transcription was measured by RT-PCR. IGF-II protein expression and clinicopathological features were assessed by immunohistochemistry and microscopic observation. The rate of IGF-II P3 methylation was significantly lower in HCC tissues (0%) than in paracancerous tissues (vs. 47.5%; x2 = 24.918, P less than 0.001) and non-cancerous tissues (vs. 100%; x2 = 80.000, P less than 0.001). IGF-II mRNA expression was significantly higher in HCC tissues (100%) than in paracancerous tissues (vs. 52.5%; x2 = 24.918, P less than 0.001) and non-cancerous tissues (vs. 0%; x2 = 80.000, P less than 0.001). IGF-II protein expression was significantly higher in HCC tissues (82.5%) than in paracancerous tissues (vs. 45.0%; x2 = 12.170, P less than 0.001) and non-cancerous tissues (vs. 0%; x2 = 56.170, P less than 0.001). IGF-II overexpression in HCC was significantly associated with degree of differentiation, extent of infiltrated serosa, size of tumor, and HBV-positive infection status. Epigenetic alterations in the IGF-II gene regulate its transcription and expression and are closely associated with HCC development and progression.

Influence of intrathecal injection of p38 MAPK inhibitor on BDNF expression in dorsal horn of spinal cord of rats with neuropathic pain / 第二军医大学学报

Objective: To observe the effect of intrathecal p38 MAPK inhibitor (SB203580) treatment on neuropathic pain and the expression of p38 MAPK and BDNF in dorsal horn of spinal cord in rats with chronic constriction injury (CCI), So as to investigate the possible mechanisms of neuropathic pain. Methods: Totally 30 SD rats were evenly randomized into 3 groups (n=10): sham group receiving intrathecal injection of sodium chloride, control group receiving intrathecal injection of sodium chloride and CCI surgery, and SB203580 group receiving intrathecal injection of SB203580 and CCI surgery. SB203580 (0.1 ml/kg) was administered 0.5 h before and 1-14 d after CCI surgery. The mechanical thresholds were tested 24 h before and 4-14 d after CCI surgery. p38 MAPK expression and BDNF release in the dorsal horn were determined using immunohistochemistry method 14 d after CCI surgery. Results: The mechanical thresholds in the control and SB203580 groups were significantly lower after CCI surgery compared with that before CCI surgery (P0.05). Compared with the sham operation group, the mechanical thresholds were significantly lower in the other two groups after CCI surgery (P<0.05). The mechanical threshold of SB203580 group was significantly higher than that of the control group after CCI surgery (P<0.05). The p38 MAPK expression and BDNF release were significantly higher in the control and SB203580 groups compared with those in the sham operation group (P<0.05), and those in the SB203580 group were significantly lower than those in the control group (P<0.05). Conclusion: Intrathecal injection of p38 MAPK inhibitor SB203580 can attenuate hyperalgesia in CCI rats through decreasing p38 MAPK expression and BDNF release.

The relationship between function and thrombosis formation of left atrium appendage assessed by transesophageal echocardiography in patients with nonvalvular atrial fibrillation / 中华全科医师杂志

Objective To explore the relationship between size,shape and function of the left atrium appendage (LAA) and its thrombosis in patients with atrial fibrillation (AF) by transesophageal echocardiography (TEE) to provide evidence for clinical risk assessment,prognosis evaluation and treatment guidance.Method Length,diameter,end-diastolic volume (EDV) and ejection velocity (PEV) of LAA were measured in 41 patients with AF,and thrombus in LAA was detected by TEE.Results Thrombus in LAA was detected in seven of 41 patients of AF (17%).No significant difference in size and EDV was found between the patients with and without thrombus,but there was significant difference in PEV between them (P