Vasodilation effect and mechanism of extraction of Tongmai Yangxin Pills (TMYX) on isolated rat mesenteric artery / 中国中药杂志

The aim of the present study is to evaluate the vasodilation effects of Tongmai Yangxin Pills (TMYX) on rat mesenteric artery as well as its mechanism of action. The relaxation effects of TMYX extracts with different concentrations were determined on isolated rat mesenteric artery in normal condition as well as pretreating by phenylephrine and KCl. Vascular relaxation effects of TMTX were also determined in mesenteric artery preincubated with L-ANME and indomethacin or in endothelium denuded mesenteric artery. Moreover, effects of TMYX by 50 mg·L⁻¹ on NO secretion and the phosphorylation of eNOS in a cellular model of human umbilical vein endothelial cell (HUVEC) pretreated with or without L-NAME were also observed. The experimental results showed that TMYX has no obvious effect on vasodilation of arteries in normal or KCl pretreated condition, while it can dose-dependently relax the rat mesenteric artery with intact endothelium stimulated with phenylephrine at a maximal diastolic rate of (64.71±10.03)%. After preincubating with L-NAME for 15 min or removal of mesenteric artery endothelium, the maximal diastolic rate was decreased to (35.77±8.93)% and (25.85±10.84)% respectively. However, preincubating with indomethacin had no inhibitory effect on TMYX induced vascular relaxation. Meanwhile, TMYX at 50 mg·L⁻¹ could increase the expression of P-eNOS and the secretion of NO in HUVEC. L-NAME significantly inhibited NO release and phosphorylation of eNOS induced by TMYX. The results suggested TMYX exerted endothelium-dependent relaxation effects against PE-induced contractions of isolated rat mesenteric artery through NO-cGMP signaling pathway.

Expression of astrocyte elevated gene-1 protein and its clinical significance in laryngeal squamous cell carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To assess the protein expression of astrocyte elevated gene-1 (AEG-1) in tissue specimens of laryngeal squamous cell carcinoma (LSCC), and to correlate its expression with clinicopathological parameters and prognosis in patients with LSCC.</p><p><b>METHODS</b>RT-PCR was used to assay the expression of AEG-1 mRNA in 13 pairs of LSCC tissues and their corresponding noncarcinoma epithelia. Immunohistochemistry was performed on paraffin-embedded tissue specimens to investigate the protein expression of AEG-1 in 88 cases of LSCC specimens and 15 cases of adjacent epithelial samples.</p><p><b>RESULTS</b>The expression of AEG-1 mRNA was significantly increased in LSCC tissues compared to adjacent noncarcinoma epithelial tissues (0.81 ± 0.17 vs. 0.23 ± 0.10;t = 10.337, P < 0.001). Meantime, the positive rate of AEG-1 protein in 88 cases of LSCC was 87.5% (77/88). However, 15 cases of adjacent noncarcinoma epithelial merely demonstrated negative or mild expression of AEG-1 protein. AEG-1 overexpression was closely correlated with T stage (χ(2) = 6.289, P = 0.018), clinical stage (χ(2) = 11.049, P < 0.01), metastasis (χ(2) = 20.859, P < 0.01) and recurrence(χ(2) = 13.459, P < 0.01). The overall survival rates of patients with AEG-1 overexpression and low expression were 35.9% and 86.4%, respectively (χ(2) = 23.409, P < 0.01). Multivariate Cox regression analysis revealed that AEG-1 expression was an independent prognostic factor (P = 0.016).</p><p><b>CONCLUSION</b>AEG-1 protein may play a critical role in the initiation and progression of LSCC, implicating its predictive value in prognosis.</p>

EphA2 promotes angiogenesis and metastasis of head and neck squamous cell carcinoma in vivo / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of EphA2 on the angiogenesis and cervical lymph node metastasis of squamous cell carcinoma of the head and neck (SCCHN) in vivo.</p><p><b>METHODS</b>EphA2 short hairpin (shRNA) lentiviral particles were used to knockdown the expression of EphA2 in SCCHN cell line M2 with high lymph nodes metastasis rate. Stable clones, obtained by puromycin screening, were assayed by RT-PCR and Western blot to validate the gene silencing efficiency and were used to establish SCCHN metastatic xenograft mouse model. Hematoxylin-eosin staining was applied to identify cervical lymph node metastasis of SCCHN in xenografted tumors. Immunohistochemistry was used to observe microvessel density. Western blot was used to investigate the protein expressions of EphA2 and vascular endothelial, growth factor (VEGF).</p><p><b>RESULTS</b>EphA2 shRNA lentiviral particles efficiently decreased the mRNA and protein expressions of EphA2 in SCCHN cell line M2, which were further successfully utilized to establish SCCHN metastatic xenograft mouse model. Compared with xenografted tumors in control group, xenografted tumors in M2EphA2RNAi(+) group decreased significantly tumor volume [(430.7 ± 190.0) mm(3) (x(-) ± s) vs (1179.0 ± 289.4) mm(3)] and weight [(0.26 ± 0.10) g vs (0.54 ± 0.12) g] (both P < 0.05). More importantly, bilateral cervical lymph node metastasis rate in M2EphA2RNAi(+) was also greatly declined (Mann-Whitney U = 10.0, P < 0.05). Decreased protein expressions of EphA2 and VEGF and microvessel density were observed in M2EphA2RNAi(+) group (t = 26.751, P < 0.01).</p><p><b>CONCLUSIONS</b>Knockdown of EphA2 expression led to the inhibition of tumor growth and metastasis in SCCHN nude mouse model. More importantly, SCCHN angiogenesis was also impeded, which might be associated with the decreased expression of VEGF.</p>

Expression of HMGB1 protein in laryngeal squamous cell carcinoma and its clinical significance / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the expression of HMGB1 protein in tissue specimens of laryngeal squamous cell carcinoma (LSCC) and adjacent normal mucosa, and explore the correlation of HMGB1 protein expression with clinicopathologic features and prognosis in LSCC.</p><p><b>METHODS</b>Ninty-three cases of LSCC and 5 cases of adjcent mucosal tissue samples were included in this study. Immunohistochemical staining was performed on paraffin-embedded tissue specimens to examine the HMGB1 protein expression. The data were futher correlated with the clinicopathological features and prognosis of the LSCC patients.</p><p><b>RESULTS</b>The positive rates of HMGB1 expression in LSCC specimens was 87.1%, significantly higher than that in the adjcent normal mucosa samples (46.7%, P = 0.001), and its overexpresion was closely correlated with T stage (Chi2 = 10.878, P = 0.004), clinical stage (Chi2 = 21.115, P < 0.01), metastasis (Chi2 = 28.298, P < 0.01) and recurrence (Chi2 = 14. 923, P = 0.001) in patients with LSCC. Patients with HMGB1 overexpression had both poorer disease-free survival and poorer overall survival compared with that in patients with low HMGB1 expression (Chi2 = 13.815, Chi2 = 11.912; Both P < 0.01). Univariate and multivariate Cox regression analyses revealed that HMGBI expression is an independent prognostic factor for patients with LSCC.</p><p><b>CONCLUSIONS</b>The results of this study demonstrate that HMGB1 protein expression is significantly increased in LSCC tissues, and HMGB1 protein overexpression is associated with a poorer prognosis in patients with LSCC. These results suggest that HMGB1 may play a critical role in the initiation and progression of LSCC, implicating HMGB1 may become a valuable marker for the prediction of prognosis in patients with LSCC.</p>

Correlation of EphA2 protein expression with clinicopathological characteristics and prognosis in laryngeal squamous cell carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the expression of EphA2 protein in tissue specimens and cell lines of laryngeal squamous cell carcinoma (LSCC), and to further study the correlation of EphA2 protein expression with clinicopathological characteristics and prognosis in LSCC.</p><p><b>METHODS</b>Western blot was applied to assess the EphA2 protein expression in LSCC cell line Hep-2 cells and the head and neck immortalized epithelial cell line NP-69 cells. Immunohistochemical staining was performed on paraffin sections of 88 cases of LSCC specimens and 16 cases of adjcent normal tissue samples to investigate the EphA2 protein expression, and to futher elucidate its correlation with clinicopathological characteristics.</p><p><b>RESULTS</b>Compared with the NP-69 cells, EphA2 expression in LSCC cell line Hep-2 cells was upregulated. The positive rates of EphA2 expression in LSCC and adjcent normal tissues samples were 80.7% and 43.8%, respectively, with a significant difference between the two groups (P < 0.001). EphA2 overexpresion was closely correlated with clinical stage (I + II/III + IV, P = 0.005), metastasis (P = 0.025) and recurrence (P = 0.021) in LSCC. Furthermore, patients with EphA2 overexpression had poorer tumor-free survival and 5-year overall survival compared with that in patients with low EphA2 expression (33.3% vs. 63.2%, P = 0.003; 46.7% vs. 81.6%, P = 0.002). EphA2 expression combined with clinical stage provided a better predictive value in prognosis. Univariate and multivariate Cox regression analysis revealed that EphA2 expression is an independent prognostic factor for patients with LSCC (P = 0.019).</p><p><b>CONCLUSIONS</b>The results of this study demonstrate that EphA2 protein expression is significantly increased in LSCC tissues and cell lines, and EphA2 protein overexpression is associated with tumor recurrence, metastasis and poorer prognosis in LSCC patients. These results suggest that EphA2 may play a critical role in the initiation and progression of LSCC, implicating EphA2 as a valuable marker for the prediction of recurrence, metastasis and prognosis in LSCC.</p>

Isocitrate dehydrogenase gene mutations in acute myeloid leukemia / 中国实验血液学杂志

The purpose of this study was to identify point mutation of the isocitrate dehydrogenase gene (IDH1 and IDH2) in patients with acute myeloid leukemia(AML) and its clinical significance. 90 de novo AML patients were selected for this study, the genomic DNA was served as template, the exon4 of IDH1 and IDH2 were amplified respectively. The IDH mutation was detected by using directly sequencing method for PCR product. The results indicated that among 90 de novo AML patients, 4 patients (4.4%) showed the IDH1 gene mutation positive, and 7(7.8%) patients showed IDH2 gene mutation positive. None was found harboring both mutations, the overall rate of mutation positive of them was 12.2%. In the AML patients with IDH gene mutation positive, the rate of normal karyotype was 72.7%, which was significantly higher than that in abnormality karyotype. The CR rate in mutation positive patients was 72.7%, which seemed as if higher than that in mutation negative patients, but without statistical significance. The mutation disappeared when the patients gained CR, and reappeared in same loci after relapse occurred. It is concluded that the IDH gene point mutation appears in normal karyotype patients, especially in patients combined with NPM1 gene mutation. The IDH gene mutation may be an important target for therapy and evaluating clinical prognosis of patients with normal karyotype.