Effect of ZNF281 on proliferation of hepatocellular carcinoma cells / 中国病理生理杂志

AIM:To investigate the role of zinc finger protein 281(ZNF281)in the proliferation of hepatocel-lular carcinoma(HCC)cells.METHODS:The mRNA expression levels of ZNF281 in peripheral blood mononuclear cells from 80 cases of healthy people and 206 cases of HCC patients were determined by real-time PCR.Statistical analysis were used to illustrate the relationship between the mRNA expression levels of ZNF 281 in the peripheral blood mononuclear cells and the clinicopathologic parameters of HCC patients.Real-time PCR and Western blot were used to detect the mRNA and protein expression levels of ZNF281 in hepatoma cell lines and immortalized hepatocytes.The silencing of ZNF281 was conducted by transfection of small interfering RNA targeting ZNF281,and then the proliferation of HCC cells was analyzed by MTS assay.The DNA synthesis of HCC cells was tested by Cell-LightTMEdU Apollo?488 In Vitro Imaging Kit.The ability of colony formation of the HCC cells was measured by colony formation assay,and the ability of anchorage-indepen-dent growth was detected by soft agar test.RESULTS: The mRNA expression level of ZNF281 in the peripheral blood mononuclear cells from HCC patients was significantly increased compared with the healthy people,and the high expression level was positively correlated with tumor size,tumor stage and tumor vascular invasion.Concurrently,the expression level of ZNF281 in hepatoma cell lines was significantly higher than that in immortalized hepatocytes.More importantly,the si-lencing of ZNF281 inhibited the proliferation, DNA synthesis, colony formation and anchorage-independent growth of the HCC cells.CONCLUSION:ZNF281 promotes the proliferation of HCC cells.

Effect of moxibustion on vascular endothelial cell and expression of vascular endothelial growth factor in rats with cutaneous wound / 中国针灸

<p><b>OBJECTIVE</b>To explore the effect and action mechanism of moxibustion on healing of cutaneous wound in rats.</p><p><b>METHODS</b>Twenty-four SD rats were selected and made into linear full-thickness skin injury model. With randomized digital table, rats were randomly divided into a treatment group and a model group, 12 cases in each one. Then according to treatment time, each group was again divided into a 1d group, a 3d group and a 7d group, 4 cases in each one. The moxibustion at injured skin was applied in the treatment group, 30 min per time, once a day. Hematoxylineosin (HE) staining method was adapted to measure growth status of capillary and number of vascular endothelial cell; immunohistochemical method was used to measure the expression of vascular endothelial growth factor (VEGF).</p><p><b>RESULTS</b>The wound healing indices in the treatment 7d group were higher than those in the model 7d group on both the 4th day and 8th day after treatment (both P < 0.05). The number of capillary in the treatment 1d group and 3d group was higher than that in the model 1 d and 3 d groups (both 1 < 0.05). The number of capillary in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The number of vascular endothelial cell in the treatment 3d group was higher than that in the model 3d group (P < 0.05). The number of vascular endothelial cell in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The difference of number of vascular endothelial cell between the treatment 1d group and model 1d group was not significant (P > 0.05). Positive cells accumulated score of V EGF expression in the treatment 3d group was higher than that in the model 3d group (P < 0.05). Positive cells accumulated score of VEGF expression in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The difference of positive cells accumulated score of VEGF expression between the treatment 1d group and model 1d group was not significant (P > 0.05).</p><p><b>CONCLUSION</b>Moxibustion could improve the healing of skin wound in rats, which could be related with regulating vascular endothelial cell and VEGF in wound tissue at different time.</p>

Effect of adenoviral N-methylpurine DNA glycosylase overexpression on chemosensitivity of human osteosarcoma cells / 中华病理学杂志

<p><b>OBJECTIVE</b>The overexpression of N-methylpurine DNA glycosylase (MPG) may imbalance the DNA base excision repair (BER) to sensitize tumor cells to current DNA damage chemotherapy. In an effort to improve the efficacy of cancer chemotherapy, we have constructed adenoviral vector of MPG, to study its ability to sensitize human osteosarcoma cell HOS to DNA damage agents.</p><p><b>METHODS</b>The adenoviral infection and MPG expression, as well as enzyme activity were determined by flow cytometry, Western blot, and HEX labeled oligonucleotide-based assay respectively. The cell survival/proliferation was measured using MTS, SRB, and [(3)H] thymidine incorporation assay. Apoptosis cell death was assayed by flow cytometry after treatment using phycoerythin (PE)-conjugated Annexin V and 7-amino-actinomycin (7-AAD).</p><p><b>RESULTS</b>A 10 MOI of recombinant nonreplicating adenovirus was found to infect more than 90% of HOS cells within 24 hours by EGFP fluorescence, in which the MPG overexpression and MPG enzyme activity were also detected. The MPG overexpression HOS cells were significantly more sensitive to the DNA damage agents, including MMS, MNNG, and TMZ, with changes in the IC50 of 6.0, 4.5, and 2.5 fold respectively.</p><p><b>CONCLUSIONS</b>These data establish transient MPG overexpression as a potential therapeutic approach for increasing HOS cellular sensitivity to DNA damage agent chemotherapy.</p>

Characteristic and clinicopathologic significance of lymphangiogenesis in colorectal cancer / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the distribution patterns and proliferative activity of lymphatic vessels in colorectal carcinomas (CRC) and their relationship with tumor metastasis and disease prognosis.</p><p><b>METHODS</b>The microlymphatic density (MLD) and microvascular density in tumoral and non-tumoral areas of 96 cases of CRC were evaluated by immunohistochemistry, using monoclonal antibodies for podoplanin and CD34 respectively. The Ki-67 expression of the lymphatic and blood vessels was detected by double-labeling immunohistochemistry. The relationship between MLD and clinicopathologic features and prognosis was analyzed.</p><p><b>RESULTS</b>The lymph vessels at central and superficia1 portions of CRC often had a reticular architecture with numerous tiny and ill-defined lumina, while those at the tumor borders had large and open lumina. The MLD at tumor borders (51.2 +/- 25.5) was significantly higher than that in normal colorectal mucosa (29.4 +/- 9.0) and other portions of CRC (P < 0.01). The Ki-67 labeling index of the lymphatic lining cells at tumor borders (0.23 +/- 0.17) was significantly higher than that in other portions of CRC (P < 0.05). The MLD significantly correlated with lymphatic involvement by tumor cells, regional lymph node metastasis and distant metastasis (P < 0.01). The 5-year survival rate was also significantly lower in patients with high MLD (P < 0.05).</p><p><b>CONCLUSIONS</b>Neolymphatic vessels are commonly seen in CRC, especially at tumor borders. High MLD at tumor borders is associated with metastasis. The detection of MLD at tumor borders may thus be useful in predicting lymph node metastasis and prognosis in patients with CPC.</p>