Mouse blastocysts were exposed to doses of 0, 1 and 10 mumol/L retinoic acid (RA) for 24 h and the cytotoxic effect of RA on the mouse blastocysts in vitro was observed. FITC-labeled terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL-FITC) assay was employed to stain apoptotic cells and immunohistochemical S-P staining method was used to detect the expression of Fas protein in mouse blastocysts in vitro. The results showed that RA could induce apoptosis and increase the expression of Fas proteins of trophectoderm (TE) and inner cell mass (ICM) cells in blastocysts. Compared with the findings for the control blastocysts, exposure to RA (10 mumol/L) resulted in a more significant apoptosis and higher expression level of Fas proteins (P<0.01). It was concluded that RA could induce apoptosis, which may result in a significant reduction in the average number of total cells and the trophectoderm/inner cell mass in blastocysts and an increased expression of Fas protein, suggesting that RA had a cytotoxic effect on the growth and development of early embryos in mice.
Antigens, CD95/*biosynthesis ; Apoptosis/*drug effects ; Blastocyst/cytology ; Blastocyst/*metabolism ; Cell Culture Techniques/methods ; Cells, Cultured ; Gene Expression Regulation/*drug effects ; In Situ Nick-End Labeling ; RNA, Messenger/metabolism ; Tretinoin/*pharmacology

This study examined the roles of HLA-G in the pathogenesis, development and immune tolerance of hemangioma. From 2000 to 2007, 52 paraffin-embedded specimens (26 from males and 26 from females) of skin capillary hemangioma and 7 samples of adjacent normal skin tissues were collected. Four fresh specimens of hemangioma were also harvested. All samples were HE-stained and proliferative cell nuclear antigen (PCNA) was immunohistochemically detected by using SP method. The samples were classified into proliferative group and degenerative group according to the Mulliken criteria and the expression pattern of PCNA. SP method and quantum dots double staining were applied to detect the expression of HLA-G and PCNA in hemangioma and normal tissue samples. The expression of HLA-G was detected by RT-PCR. The results showed that among the 52 samples of hemangioma, 29 were of proliferative type and 23 degenerative type, and of the four fresh samples of hemangioma, 2 were of proliferative type and 2 degenerative type. SP method results showed that HLA-G was expressed in both proliferative and degenerative hemangioma, but not in normal tissues. The quantum dots double staining exhibited that HLA-G expression was significantly higher in proliferative group than in degenerative (P<0.05) and normal groups (P<0.05), but there was no statistically significant difference between the latter two groups (P>0.05). RT-PCR revealed that HLA-G was transcribed in both the proliferative and degenerative hemangioma tissues, but not in normal tissues. We are led to conclude that the elevated expression of HLA-G in proliferative hemangioma cells may lead to immune tolerance, which allows cells to escape immune surveillance and proliferate. On the other hand, the lower expression of HLA-G in degenerative hemangioma may result in immune cells-induced degeneration of hemangioma.

Objective To investigate the expression of survivin and its relationship with that of caspase-3 in two stages of hemangioma and normal skin, and to explore the role of survivin in the patho-genesis of hemangioma. Methods A total of 50 cases of human dermal hemangioma and 8 eases of normal skin were analyzed for expression of survivin and caspase-3 by immunohistochemistry. Results The posi-tive ratio and average light density of survivin were obviously higher in proliferative hemangioma (0.2449±0.0135,0.7246±0.0747) than that in involutional hemangioma (0.1648±0.0217,0.5592±0.1601) and normal skin (0.1789±0.0126,0.4626±0.0961) (P<0.01). On the contrary, the expression of easpase-3 was up-regulated in involutional hemangioma (0.2386±0.0175, 0.4378±0.0593). Analysis of data also showed that there was a negative correlation between the average light density of survivin and caspase-3 (r=-0.95,P<0.01). Conclusions Survivin plays an important role in the pathogenesis of hemangioma and has a negative relationship with easpase-3.

The relationship between Bcl-2, Bax, Fas, caspase-3 and development of hemangioma and the molecular mechanism was investigated. By using immunohistochemical S-P method, proliferating cell nuclear antigen was detected. According to the classification of Mulliken in combination with PCNA expression, 27 cases were identified as proliferating hemangioma and 22 cases as involutive hemangioma. Five normal skin tissues around the tumor tissue served as controls. By using immunohistochemical technique, the expression of Bcl-2, Bax, Fax and Caspase-3 was detected. The cells expressing Bcl-2, Bax, Fax and cappase-3 were identified as hemangioma endothelia by immunohistochemical staining of VIII factor. The average absorbance (A) and average positive area rate of Bcl-2, Bax, Fas and caspase-3 expression were measured by using HPIAS-2000 imaging analysis system. The results showed that the expression of Bcl-2 in the endothelia of proliferating hemangioma was significantly higher that in involutive degenerative hemangioma endothelia and vascular endothelia of normal skin tissue (P < 0.01). The expression of Bax, Fas and Caspase-3 in the endothelia of involutive hemangioma was obviously higher than in the endothelia of proliferating hemangioma and normal skin tissue (P < 0.01). The expression of BAx and Fas in endothelia of proliferating hemangioma was higher than in those of normal skin tissue (P < 0.05). It was suggested that Bcl-2, Bax, Fas and caspase-3 might be involved in the development and involution of hemangioma. Bcl-2 could promote the growth of hemangioma by inhibiting apoptosis of endothelia. Bax, Fas and caspase-3 promote the switch of hemangioma from proliferation to involution by inducing the apoptosis of hemangioma endothelia.

Objective To investigate the relationship between the expression of p63, p73 proteins and the development of hemangioma. Methods The immunohistochemical technique and quantitative image analysis were used to detect the expression of p63 and p73 proteins in 40 cases of capillary hemangioma and 20 specimens of normal skin. Results The absorbance value (mean ? SD) of p63 and p73 expression in normal skin tissue, proliferative phase of hemangioma and involuting phase of hemangioma were 0.923 ? 0.191 and 0.953 ? 0.120, 8.271 ? 1.953 and 6.408 ? 2.151, 0.920 ? 0.187 and 1.073 ? 0.516, respectively. The expression of p63 and p73 in proliferative phase of hemangioma was significantly increased as compared with those in involuting phase of hemangioma and normal skin tissue (P 0.05). Conclusions It is suggested that p63 gene is not a tumor suppressor gene but an oncogene in hemangioma and may contribute to the proliferation of endothelial cell and be associated with angiogenesis, and p73 may play an important role in the proliferation of hemangioma.

The action mechanism of matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of metalloproteinases-2 (TIMP-2) in the genesis, development and degeneration of haemangioma was investigated by detecting their expression in the tissue of haemangioma in different phases by using the immunohistochemistry. Fifty paraffin-embedded specimens of skin capillary haemangioma were collected, which were documented in the Department of Pathology, Renmin Hospital of Wuhan University from 2000 to 2006. All samples were stained by regular HE method, and proliferative cell nuclear antigen (PCNA) was tested by immunohistochemical S-P method. The samples were classified according to the Mulliken criteria and the expression pattern of PCNA. Immunohistochemical S-P method was applied to detect the expression of MMP-2 and TIMP-2 in proliferative and degenerative phases of cutaneous capillary haemangioma, and in normal skin tissues. In combination with the detection of the expression of factor VIII-related antigen, it was verified that in haemangioma tissues, the cells expressing MMP-2 and TIMP-2 were vascular endothelial cells. The MMP-2 and TIMP-2 expression was quantitatively analyzed by image analysis system (HPIAS-1000), and one-way ANOVA(107) and SNK(q) test were done to analyze average absorbance (A) and positive area rate of immunohistochemically positive particles by using SPSS11.5. The results showed: (1) Among 50 samples of haemangioma, there were 26 proliferative haemangiomas, and 24 degenerative haemangiomas, respectively; (2) The expression of MMP-2 was weak in normal vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression of MMP-2 in proliferative group was significantly higher than in degenerative group and control group (normal skin) (P<0.05), but there was no statistically significant difference between the latter two groups; (3) TIMP-2 was highly expressed in normal tissues, degenerative vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression level of TIMP-2 in proliferative phase was significantly lower than in degenerative phase (P<0.05), and the expression of TIMP-2 in proliferative phase was significantly different from that in degenerative phase and normal tissues (P<0.05). It was concluded that in proliferative phase of haemangioma, MMP-2 may promote over-proliferation of endothelial cells of haemangioma, and in degenerative phase, TIMP-2 can inhibit the proliferation of endothelial cells of haemangioma. The two substances play important roles in the genesis, development and degeneration of haemangiomas.

This study examined the roles of HLA-G in the pathogenesis, development and immune tolerance of hemangioma. From 2000 to 2007, 52 paraffin-embedded specimens (26 from males and 26 from females) of skin capillary hemangioma and 7 samples of adjacent normal skin tissues were collected. Four fresh specimens of hemangioma were also harvested. All samples were HE-stained and proliferative cell nuclear antigen (PCNA) was immunohistochemically detected by using SP method. The samples were classified into proliferative group and degenerative group according to the Mulliken criteria and the expression pattern of PCNA. SP method and quantum dots double staining were applied to detect the expression of HLA-G and PCNA in hemangioma and normal tissue samples. The expression of HLA-G was detected by RT-PCR. The results showed that among the 52 samples of hemangioma, 29 were of proliferative type and 23 degenerative type, and of the four fresh samples of hemangioma, 2 were of proliferative type and 2 degenerative type. SP method results showed that HLA-G was expressed in both proliferative and degenerative hemangioma, but not in normal tissues. The quantum dots double staining exhibited that HLA-G expression was significantly higher in proliferative group than in degenerative (P<0.05) and normal groups (P<0.05), but there was no statistically significant difference between the latter two groups (P>0.05). RT-PCR revealed that HLA-G was transcribed in both the proliferative and degenerative hemangioma tissues, but not in normal tissues. We are led to conclude that the elevated expression of HLA-G in proliferative hemangioma cells may lead to immune tolerance, which allows cells to escape immune surveillance and proliferate. On the other hand, the lower expression of HLA-G in degenerative hemangioma may result in immune cells-induced degeneration of hemangioma.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; HLA-G Antigens ; genetics ; Hemangioma, Capillary ; genetics ; Humans ; Infant ; Male ; Middle Aged ; Skin Neoplasms ; genetics ; Young Adult

Objective To investigate the key procedures of the acute traumatic intracranial hematoma com-bined with herniation and the prognosis factors. Methods 45 cases of acute traumatic intraeranial hematoma com-bined with herniation from February 1997 to June 2008 were admitted in our hospital. Timely establishment of effec-tive ventilation and circulation and pre-operative examination were done to all the eases. Craniotomy hematoma clean was performed in 8 cases, hematoma clean and decompressive craniectomy was canducted in 33 cases and 4 cases were not operatively treated. Results 26 eases (58%) were cured,and 19 cases (42%) died. Conclusions The key procedures of the acute tranmatie intraeranial hematoma combined with herniation is timely establishment of ef-fective ventilation and circulation, and that is effective method to prevent secondary brain injury ; removing hematoma as soon as possible,and lifting the oppression of the brain stem are the keys to rescue patients. Prognosis is closely related to the degree of primary brain injury, eonseious level before operation and the time of herniation appearance.

Objective To investigate the function of Bcl-2 and Bax in the pathogenesis,development and regression of human hemangiomas.Methods We examined the expression of Bcl-2 and Bax in proliferating versus involuting human hemangioma tissues and normal skin tissues using immunohistochemical technique.Results The expression of Bcl-2 in proliferating hemangiomas was significantly higher than that in involuting hemangiomas and normal skin tissues(P＜0.01).No significant difference was found between the expression of Bcl-2 in involuting hemangiomas and that in normal skin tissues(P＞0.05).The expression of Bax in involuting hemangiomas was significantly higher than that in proliferating hemangiomas and normal skin tissues(P＜0.01);the expression of Bax in proliferating hemangiomas was significantly higher than that in normal skin tissues(P＜0.05).Conclusion Bcl-2 and Bax participate in the development and involution of hemangioma,Bcl-2 plays a role in accelerating the proliferation of hemangioma by inhibiting the apoptosis of endothelial cells,and Bax promotes the switching from proliferation to involution in hemangiomas through inducing the apoptosis of endothelial cells.