Clinical efficacy of propranolol in the treatment of hemangioma and changes in serum VEGF, bFGF and MMP-9.

The aim of this study was to observe the clinical efficacy and safety of propranolol in the treatment of hemangioma, and to reveal its possible mechanism. A total of 129 cases of proliferative hemangioma were divided into two groups: i) Treatment (n=97), in which the patients received oral propranolol therapy and ii) observation (n=32), in which the patients underwent clinical observation. The changes in the hemangiomas were noted and compared between the two groups. In addition, the heart rate, blood glucose levels, liver, kidney and thyroid function of the patients in the treatment group were monitored prior to and following treatment; the ELISA method was used for the measurement of the patients' serum concentrations of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and matrix metalloproteinase-9 (MMP-9) at the same time-points. A significant difference in curative effect was found between the treatment and observation group. The concentration of free thyroxine and sensitive thyroid-stimulating hormone and the heart rate of the treatment group exhibited significant changes prior to and following medication, but no statistical significance was found in the changes in blood glucose, liver and kidney function and free triiodothyronine concentration. Furthermore, the serum concentrations of VEGF, bFGF and MMP-9 in the treatment group 8 weeks after medication were decreased significantly compared with those before treatment. In conclusion, oral propranolol has a good curative effect in the treatment of proliferative hemangioma, with few side effects and a high level of safety. The mechanism underlying the effects of propranolol may be associated with the downregulation of VEGF, bFGF and MMP-9 expression.

[The application of medpor and split-thickness skin graft in formation of cranioauricular sulcus during auricular reconstruction with Nagata method].

OBJECTIVE: To investigate the application of medpor and split-thickness skin graft in formation of cranioauricular sulcus during auricular reconstruction with Nagata method. METHODS: The first stage operation was fulfilled according to the Nagata two-stage method which involves fabrication and grafting of the costal cartilage framework. The second-stage ear elevation operation was undertaken 6 months later to form the cranioauricular sulcus. Split-thickness skin was taken from temporal and accipital area. After releasing the auricular framework and transplanting C shaped medpor at the rear side of framework, the temporaparietal fascia flap was transferred to cover postauricular medpor and framework. Then the split-thickness skin graft was implant on the fascia surface. RESULTS: From July 2010 to August 2012, 20 cases (22 ears) were treated. Partial necrosis of temporaparietal fascia flap and framework exposure happened in 1 case. Successful ear reconstruction was achieved in other cases with satisfactory cranioauricular sulcus during the follow-up period of 6-18 months (average, 13 months). CONCLUSIONS: The application of medpor and split-thickness skin graft in the ear elevation of Nagata method for auricular reconstruction for microtia can achieve satisfactory results. It not only avoids the obvious scar in the donor site due to harvesting full-thickness and intermediate-thickness skin, but also reduces chest trauma due to harvesting costal cartilage.

[Auricular reconstruction for concha-type microtia].

OBJECTIVE: To investigate the method of auricular reconstruction for concha-type microtia. METHODS: Two-staged auricular reconstruction was applied in 13 cases (14 ears) with concha-type microtia. The cartilage auricular framework was fabricated and implanted in the first stage, followed by ear elevation and cranio-auricle angle formation at the second stage. RESULTS: The patients were followed up for 2 months to 2 years with satisfactory aesthetic result. The reconstructed ears had a good appearance and position, and were symmetric to the healthy ears. CONCLUSIONS: The two-staged auricular reconstruction with autologous cartilage framework is ideal for concha-type microtia.

[Construction of recombinant lentiviral vector of Tie2-RNAi and its influence on malignant melanoma cells in vitro].

OBJECTIVE: To construct lentivector carrying Tie2-Small interfering RNA (SiRNA), so as to study its influence on malignant melanoma cells. METHODS: Recombinant plasmid pSilencer 1.0-U6-Tie2-siRNA and plasmid pNL-EGFP were digested with XbaI, ligated a target lentiviral transfer plasmid of pNL-EGFP-U6-Tie2-I or pNL-EGFP-U6-Tie2-II, and then the electrophoresis clones was sequenced. Plasmids of pNL-EGFP-U6-Tie2-I and pNL-EGFP-U6-Tie2-II were constructed and combined with pVSVG and pHelper, respectively, to constitute lentiviral vector system of three plasmids. The Lentiviral vector system was transfected into 293T cell to produce pNL-EGFP-U6-Tie2- I and pNL-EGFP-U6-Tie2-II lentivirus. Then the supernatant was collected to determine the titer. Malignant melanoma cells were infected by both lentiviruses and identified by Realtime RT-PCR to assess inhibitory efficiency. RESULTS: The recombinant lentiviral vectors of Tie2-RNAi were constructed successfully which were analyzed with restriction enzyme digestion and identified by sequencing. And the titer of lentiviral vector was 8.8 x 10(3)/ml, which was determined by 293T cell. The results of Realtime RT-PCR demonstrated that the lentiviral vectors of Tie2-RNAi could infect malignant melanoma cells and inhibit the expression of Tie2 genes in malignant melanoma cells (P<0.01). There was no significant difference in the expression level (P>0.05) between the two lentiviral vectors of Tie2-RNAi. CONCLUSIONS: Lentivector carrying Tie2-SiRNA can be constructed successfully and inhibit the expression of Tie2 gene in vitro significantly. The study will supply the theory basis for the further research on the inhibition of tumor growth in vivo.

[Effectiveness of two-stage operation of auricular reconstruction in treatment of lobule-type microtia].

OBJECTIVE: To investigate the method and effectiveness of two-stage operation of auricular reconstruction in treating lobule-type microtia. METHODS: Between March 2007 and April 2010, 19 patients (19 ears) of lobule-type microtia were treated. There were 13 males and 6 females, aged 5 to 27 years (mean, 12.6 years). Of 19 patients, 11 were less than or equal to 14 years old. The locations were left ear in 9 cases and right ear in 10 cases. Two-stage operation for auricular reconstruction of lobule-type microtia included fabrication and grafting of the costal cartilage framework at the first-stage operation and the ear elevation operation at the second-stage operation. RESULTS: Pseudomonas aeruginosa infection occurred in 1 patient after the first-stage operation, who was not given the second-stage operation. Skin necrosis occurred in 1 patient 8 days after the second-stage operation and healed after symptomatic treatment. Eighteen patients were followed up 6 months to 2 years (mean, 14 months). Retraction of cranioauricular angle and thoracic deformity occurred in 1 patient. The surgical results were satisfactory in the other 17 patients whose reconstructive ear had verisimilar shape and suitable cranioauricular angle. CONCLUSION: Two-stage operation of auricular reconstruction is considered to be an ideal method for lobule-type microtia.

[Experimental study of repairing skull defect with autogenous cranial bone dust].

OBJECTIVE: To build animal model of skull defect for the basic research of skull defects reconstruction with autogenous cranial bone dust and its relevance to the clinical application. METHODS: Three full-thickness parietal skull defects (A, B and C) were created in 30 New Zealand white rabbits. The size of all the defects was 1 cm in diameter. The defect A was left untreated as control. Defects B and C were reconstructed with autogenous cranial bone dust. Two pieces of pyroxylin membrane were placed on the top and bottom of the defect B. every 10 rabbits were killed for analysis at 4, 8, 12 weeks after operation. RESULTS: Defect A was largely repaired with connective tissue. Defect B was repaired rapidly with newly formed cancellous bone in the early period, but the following process of the growth, remodeling and maturing of the newly-formed cancellous bone was slowly. The bone ingrowth in defect C was more mature, but could not repair the defect completely, especially in the central zone. The grafted bone dust was absorbed gradually. Active angiogenesis could be observed in the newly formed bone. For the same defect, new bone had a greater amount of calcium at 12 weeks than at 4 and 8 weeks after operation (P < 0.05). And the calcium content of new bone was higher in defect C than in defect B at 8, 12 weeks after operation (P < 0.05). CONCLUSIONS: The osteogenesis and angiogenesis are closely related to the time and location. The pyroxylin membrane can significantly promote the formation of cancellous bone in defect with autogenous bone dust graft during the early period.

[Estrogen on stimulating the angiogenesis of children's hemangioma in vitro].

OBJECTIVE: To demonstrate that estrogen stimulates the angiogenesis of children' s hemangioma. METHODS: A piece of hemangioma biopsy was embedded in fibrin gel, and a model in vitro of angiogenesis of human hemangioma was then established. The angiogenesis of hemangioma in each group was interfered by the estrogen and tamoxifen. There were four groups divided into the followings: the group with estrogen, the group with tamoxifen, the group with estrogen + tamoxifen and the control. The dimension of newborn tubule area in the 3rd, 6th, 9th day after the culture was calculated to compare statistically differences among the groups. RESULTS: In the model of angiogenesis of hemangioma, microvessels grew out from the tissue sample in 2 to 3 days after the culture, and in 8 to 9 days a complex network of microvessels had been shown, the tending to inactivity. On the 3rd,6th and 9th day after the culture the dimension of newborn tubule area of the group of estrogen [(2.84 +/- 0.20) mm2 (12.93 +/- 0.85) mm2 (22.47 +/- 1.40) mm2] were larger than those of the control [(1.98 +/- 0.17) mm2, (7.51 +/- 0.48) mm2, (11.26 +/- 0.73) mm2]. Those of the group of estrogen + tamoxifen [(1.08 +/- 0.11) mm2, (3.54 +/- 0.31) mm2, (5.72 +/- 0.40 mm2] and the group of tamoxifen [(1.13 +/- 0.14) mm2 (4.26 +/- 0.29) mm2, (6.08 +/- 0.42) mm2] were smaller than those of the groups of the estrogen and the control (P < 0.05). CONCLUSIONS: The estrogen may stimulate the angiogenesis of children's hemangioma, and the tamoxifen may reverse the process.

[Expression and significance of Ang1, Ang2 and receptor Tie2 in hemangioma].

OBJECTIVE: To investigate the relationship of angiogenesis and the Ang family members/ receptor (Ang/Tie2) in hemangioma. METHODS: Expression of Ang1, Ang2 and the receptor Tie2 was detected with immunohistochemical SP method and RT-PCR method in 17 cases of proliferating hemangioma, 13 involuting cases and 10 cases of normal children skin. RESULTS: The expression of Ang2 and Tie2 was higher markedly in proliferating hemangiomas than in involuting hemangiomas (P < 0.01), and was rare or negative in normal skin. Expression of Ang1 was rare or negative both in hemangioma and normal skin without significant difference between them (P > 0.05). CONCLUSION: Ang/Tie2 system may play an important role in the proliferating and involuting process of hemangioma.

[The reconstruction of funnel chest deformity with the reversed transplantation of sternum-costicartilage flap carried by the abdominal rectus pedicle].

OBJECTIVE: To investigate the reconstructive operative procedures of funnel chest with "sternum-costicartilage" flap carried by the abdominal rectus pedicle. METHODS: (1) In accordance with the lesioned area of funnel-like depressed deformity of anterior thoracic wall, a perpendicular median incision was designed and made; (2) The "sternum-costicartilage" flap carrying the abdominal rectus pedicle was used and reversed and transplanted to reconstruct severe funnel chest deformity. RESULTS: The procedure was used in 7 cases from 1999 to 2005. The results of surgery were satisfactory. There were no recurrence after operation. CONCLUSIONS: The procedure reported here is rather safe, solid and sound with good therapeutic results, and is of great value in clinical practice.

[A three-dimension in vitro model for angiogenesis of hemangioma].

OBJECTIVE: To create a three dimension (3D) in vitro model for angiogenesis of hemangioma. METHOD: The fragment of hemangioma specimen was embedded in fibrin gel to set up the three-dimension (3D) in vitro model for angiogenesis of hemangioma. RESULT: In the model, microvessels grew out from the tissue fragments at the 2nd to 3rd day after culture, and at the 8th to 9th day a compact network of microvessels come into being, then tending to be stationary. The compact network around the tissue fragment was confirmed to be blood vessels by immunohistochemistry and electron microscopy. CONCLUSION: This model helps to study the mechanism of hemangioma angiogenesis and investigate the drugs of anti-angiogenesis.

[Expression of apoptosis and Bcl-2, Bax in hemangioma and vascular malformations].

OBJECTIVE: To study the expression of apoptosis and their regulatory genes, Bcl-2, Bax in hemangioma and vascular malformations. METHODS: The specimens were taken from 68 cases of strawberry hemangioma or vascular malformations and 11 cases of normal skin. The expression of Bcl-2, Bax and Ki-67 in endothelial cells were investigated by immunohistochemical S-P method. The apoptosis index (AI) in endothelial cells was investigated by TUNEL method. RESULTS: Ki-67 was positive in all the proliferating strawberry hemangioma, negative in the other groups. The expression of Bax was significantly higher in strawberry hemangioma than in vascular malformations and normal skin (P < 0.01). Bcl-2 was negative in all groups. The AI in endothelial cells of strawberry hemangioma was significantly higher than that of vascular malformations and normal skin. There were no statistically significant differences between the proliferating and involution strawberry hemangioma. There were no statistically significant differences between various types of vascular malformations and normal skin. With the increasing of the expression of Bax in strawberry hemangioma, the AI increased. CONCLUSION: These findings clearly demonstrate a much higher apoptotic activity in strawberry hemangioma than in vascular malformations and normal skin and suggest that apoptosis might be a cause of spontaneous involution of strawberry hemangioma and might not be a cause of the pathogenesis of vascular malformation. Bcl-2, Bax might play an important regulatory role in apoptosis in endothelial cells of strawberry hemangioma.