OBJECTIVETo 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.

METHODSThree 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.

RESULTSDefect 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).

CONCLUSIONSThe 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.

Animals ; Bone Regeneration ; Bone Substitutes ; Bone Transplantation ; Female ; Male ; Materials Testing ; Osteogenesis ; Rabbits ; Skull ; Transplantation, Autologous

OBJECTIVETo study the expression of apoptosis and their regulatory genes, Bcl-2, Bax in hemangioma and vascular malformations.

METHODSThe 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.

RESULTSKi-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.

CONCLUSIONThese 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.

Adolescent ; Adult ; Apoptosis ; Child ; Child, Preschool ; Endothelial Cells ; chemistry ; pathology ; ultrastructure ; Hemangioma ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Infant ; Ki-67 Antigen ; analysis ; Microscopy, Electron ; Middle Aged ; Proto-Oncogene Proteins ; biosynthesis ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; bcl-2-Associated X Protein

OBJECTIVETo investigate the relationship of angiogenesis and the Ang family members/ receptor (Ang/Tie2) in hemangioma.

METHODSExpression 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.

RESULTSThe 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).

CONCLUSIONAng/Tie2 system may play an important role in the proliferating and involuting process of hemangioma.

Angiopoietin-1 ; metabolism ; Angiopoietin-2 ; metabolism ; Child, Preschool ; Hemangioma ; metabolism ; pathology ; Humans ; Infant ; Receptor, TIE-2 ; metabolism

OBJECTIVETo construct lentivector carrying Tie2-Small interfering RNA (SiRNA), so as to study its influence on malignant melanoma cells.

METHODSRecombinant 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.

RESULTSThe 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.

CONCLUSIONSLentivector 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.

Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Melanoma ; genetics ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, TIE-2 ; genetics ; Transfection

OBJECTIVETo investigate the method of auricular reconstruction for concha-type microtia.

METHODSTwo-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.

RESULTSThe 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.

CONCLUSIONSThe two-staged auricular reconstruction with autologous cartilage framework is ideal for concha-type microtia.

Adolescent ; Adult ; Cartilage ; transplantation ; Child ; Ear Auricle ; surgery ; Ear, External ; abnormalities ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; Ribs ; Tissue Scaffolds ; Transplantation, Autologous ; Young Adult

OBJECTIVETo 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.

RESULTSThe procedure was used in 7 cases from 1999 to 2005. The results of surgery were satisfactory. There were no recurrence after operation.

CONCLUSIONSThe procedure reported here is rather safe, solid and sound with good therapeutic results, and is of great value in clinical practice.

Bone Transplantation ; Child ; Child, Preschool ; Female ; Funnel Chest ; surgery ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Rectus Abdominis ; transplantation ; Ribs ; transplantation ; Sternum ; transplantation ; Surgical Flaps

OBJECTIVETo create a three dimension (3D) in vitro model for angiogenesis of hemangioma.

METHODThe fragment of hemangioma specimen was embedded in fibrin gel to set up the three-dimension (3D) in vitro model for angiogenesis of hemangioma.

RESULTIn 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.

CONCLUSIONThis model helps to study the mechanism of hemangioma angiogenesis and investigate the drugs of anti-angiogenesis.

Cells, Cultured ; Endothelium, Vascular ; Hemangioma ; Humans ; Models, Cardiovascular ; Neovascularization, Pathologic

OBJECTIVETo demonstrate that estrogen stimulates the angiogenesis of children' s hemangioma.

METHODSA 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.

RESULTSIn 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).

CONCLUSIONSThe estrogen may stimulate the angiogenesis of children's hemangioma, and the tamoxifen may reverse the process.

Child ; Estrogens ; adverse effects ; Hemangioma ; pathology ; Humans ; In Vitro Techniques ; Neovascularization, Pathologic ; pathology

Objective:To analyze the factors affecting delayed chemotherapy in children with Burkitt lymphoma (BL) and their influence on prognosis.Methods:Retrospective cohort study. Clinical data of 591 children aged ≤18 years with BL from May 2017 to December 2022 in China Net Childhood Lymphoma (CNCL) was collected. The patients were treated according to the protocol CNCL-BL-2017. According to the clinical characteristics, therapeutic regimen was divided into group A, group B and group C .Based on whether the total chemotherapy time was delayed, patients were divided into two groups: the delayed chemotherapy group and the non-delayed chemotherapy group. Based on the total delayed time of chemotherapy, patients in group C were divided into non-delayed chemotherapy group, 1-7 days delayed group and more than 7 days delayed group. Relationships between delayed chemotherapy and gender, age, tumor lysis syndrome before chemotherapy, bone marrow involvement, disease group (B/C group), serum lactate dehydrogenase (LDH) > 4 times than normal, grade Ⅲ-Ⅳ myelosuppression after chemotherapy, minimal residual disease in the interim assessment, and severe infection (including severe pneumonia, sepsis, meningitis, chickenpox, etc.) were analyzed. Logistic analysis was used to identify the relevant factors. Kaplan-Meier method was used to analyze the patients' survival information. Log-Rank was used for comparison between groups.Results:Among 591 patients, 504 were males and 87 were females, the follow-up time was 34.8 (18.6,50.1) months. The 3-year overall survival (OS) rate was (92.5±1.1)%,and the 3-year event-free survival (EFS) rate was (90.5±1.2)%. Seventy-three (12.4%) patients were in delayed chemotherapy group and 518 (87.6%) patients were in non-delayed chemotherapy group. The reasons for chemotherapy delay included 72 cases (98.6%) of severe infection, 65 cases (89.0%) of bone marrow suppression, 35 cases (47.9%) of organ dysfunction, 22 cases (30.1%) of tumor lysis syndrome,etc. There were 7 cases of chemotherapy delay in group B, which were seen in COPADM (vincristine+cyclophosphamide+prednisone+daunorubicin+methotrexate+intrathecal injection,4 cases) and CYM (methotrexate+cytarabine+intrathecal injection,3 cases) stages. There were 66 cases of chemotherapy delay in group C, which were common in COPADM (28 cases) and CYVE 1 (low dose cytarabine+high dose cytarabine+etoposide+methotrexate, 12 cases) stages. Multinomial Logistic regression analysis showed that the age over 10 years old ( OR=0.54,95% CI 0.30-0.93), tumor lysis syndrome before chemotherapy ( OR=0.48,95% CI 0.27-0.84) and grade Ⅲ-Ⅳ myelosuppression after chemotherapy ( OR=0.55,95% CI 0.33-0.91)were independent risk factors for chemotherapy delay.The 3-year OS rate and the 3-year EFS rate of children with Burkitt lymphoma in the delayed chemotherapy group were lower than those in the non-delayed chemotherapy group ((79.4±4.9)% vs. (94.2±1.1)%, (80.2±4.8)% vs. (92.0±1.2)%,both P<0.05). The 3-year OS rate of the group C with chemotherapy delay >7 days (42 cases) was lower than that of the group with chemotherapy delay of 1-7 days (22 cases) and the non-delay group (399 cases) ((76.7±6.9)% vs. (81.8±8.2)% vs. (92.7±1.3)%, P=0.002).The 3-year OS rate of the chemotherapy delay group (9 cases) in the COP (vincristine+cyclophosphamide+prednisone) phase was lower than that of the non-chemotherapy delay group (454 cases) ((66.7±15.7)% vs. (91.3±1.4)%, P=0.005). Similarly, the 3-year OS rate of the chemotherapy delay group (11 cases) in the COPADM1 phase was lower than that of the non-chemotherapy delay group (452 cases) ((63.6±14.5)% vs. (91.5±1.3)%, P=0.001). Conclusions:The delayed chemotherapy was related to the age over 10 years old, tumor lysis syndrome before chemotherapy and grade Ⅲ-Ⅳ myelosuppression after chemotherapy in pediatric BL. There is a significant relationship between delayed chemotherapy and prognosis of BL in children.

Objective To explore the mechanism of the intestinal barrier damage caused by Blastocystis hominis infections in rats. Methods Thirty SD rats were randomly divided into the control group, and the 1-, 3-, 6- and 9-week-infection groups, of 6 rats in each group. Rats in each infection group were orally infected with B. hominis trophozoites at a density of 2 × 10⁸ parasites per rat, and the control group was given an equal volume of phosphate buffered saline solution. The 7-hour urine samples were collected 1, 3, 6 and 9 weeks post-infection for the measurement of the intestinal permeability. Then, rats were sacrificed using the cervical dislocation method, and the cecum specimens were collected for the detection of the intestinal epithelial cell permeability. The expression of tight junction-related Occludin and Claudin - 1 genes and apoptosis-related Bcl - 2 and Bax genes was quantified in cecum epithelial cells using the real-time fluorescent quantitative PCR (qPCR) assay, and cell apoptosis was detected in the rat cecum using the TdT-mediated dUTP nick-end labeling (TUNEL) assay. Results The median urinary lactolose to mannitol ratios were 0.29, 0.72, 0.44, 0.46 and 0.38 in the control group, and the 1-, 3-, 6- and 9-week-infection groups, respectively, and the difference was statistically significant (H = 12.09, P < 0.05). B. hominis invasion and epithelial injury were observed in intestinal epithelial cells of rats infected with B. hominis, and transmission electron microscopy displayed the destruction of tight junctions between intestinal epithelial cells. The relative expression of Occludin, Claudin-1, Bcl-2 and Bax genes was 1.04, 0.62, 0.71, 0.68 and 0.96; 1.03, 0.61, 0.63, 0.76 and 0.86; 1.08, 0.70, 0.75, 0.74 and 1.03; and 1.00, 1.57, 1.33, 1.35 and 1.10 in the control group and the 1-, 3-, 6- and 9-week-infection groups, respectively, and all differences were statistically significant (F = 2.86, 2.85, 3.37 and 4.45, all P values < 0.05). The median number of positive staining cells were 1.00, 13.00, 9.00, 3.50 and 1.00 in rat cecum specimens in the control group, and the 1-, 3-, 6- and 9-week-infection groups, respectively, and the difference was statistically significant (H = 22.95, P < 0.01). Conclusion B. hominis infection may cause an increase in the rat intestinal permeability through triggering the apoptosis of intestinal epithelial cells to destroy the tight junction between intestinal epithelial cells, thereby destroying the intestinal barrier function.