Objective:To investigate the application and effect of frontal-orbital reconstruction and distraction osteogenesis in the treatment of Crouzon syndrome using three-dimensiond reconstruction mold (3D-RM).Methods:Retrospective analysis was performed on the clinical data of children with Crouzon syndrome who were treated in the Department of Burning and Plastic Surgery, Children’s Hospital of Nanjing Medical University from January 2018 to December 2021. These children were divided into two groups. The study group performed fronto-orbital reconstruction combined with distraction osteogenesis using 3D-RM, while the control group performed the surgery without using 3D-RM. Intraoperative blood loss, operation time, duration of hospital stay, complication rate, cranial index, frontal and orbital retraction rate and postoperative frontal and orbital contour were compared between the two groups. The independent sample t-test was used to compare the differences in intraoperative blood loss, operation duration, hospital stay, postoperative cranial index and fronto-orbital retraction rate between the two groups. The Chi-square test was used to compare the differences in the incidence of postoperative complications between the two groups. Results:The study group included 10 children, including 6 males and 4 females, ranging in age from 9 to 37 months, with an average age of 19 months. In the control group, 8 children were included, including 5 males and 3 females, ranging in age from 8 to 46 months, with an average age of 25 months. The study group was followed up for 12 to 36 months, with an average of 18 months, and the head shape was better improved. The control group was followed up for 12 to 30 months, with an average of 18 months. No serious complications occurred and the head deformity was improved. There were no significant differences in intraoperative blood loss, operation duration, hospital stay, cranial index, fronto-orbital retraction rate and complication rate between the two groups (all P>0.05). The forehead shape of the study group was more symmetrical than that of the control group, the difference was statistically significant ( P<0.05). Conclusion:The effect of frontal orbital reconstruction and distraction osteogenesis using 3D-RM in the treatment of Crouzon syndrome is positive, the head shape and orbit are significantly improved. There were no serious complications after operation. It can play a good auxiliary role in the operation.

Objective:To investigate the application and effect of frontal-orbital reconstruction and distraction osteogenesis in the treatment of Crouzon syndrome using three-dimensiond reconstruction mold (3D-RM).Methods:Retrospective analysis was performed on the clinical data of children with Crouzon syndrome who were treated in the Department of Burning and Plastic Surgery, Children’s Hospital of Nanjing Medical University from January 2018 to December 2021. These children were divided into two groups. The study group performed fronto-orbital reconstruction combined with distraction osteogenesis using 3D-RM, while the control group performed the surgery without using 3D-RM. Intraoperative blood loss, operation time, duration of hospital stay, complication rate, cranial index, frontal and orbital retraction rate and postoperative frontal and orbital contour were compared between the two groups. The independent sample t-test was used to compare the differences in intraoperative blood loss, operation duration, hospital stay, postoperative cranial index and fronto-orbital retraction rate between the two groups. The Chi-square test was used to compare the differences in the incidence of postoperative complications between the two groups. Results:The study group included 10 children, including 6 males and 4 females, ranging in age from 9 to 37 months, with an average age of 19 months. In the control group, 8 children were included, including 5 males and 3 females, ranging in age from 8 to 46 months, with an average age of 25 months. The study group was followed up for 12 to 36 months, with an average of 18 months, and the head shape was better improved. The control group was followed up for 12 to 30 months, with an average of 18 months. No serious complications occurred and the head deformity was improved. There were no significant differences in intraoperative blood loss, operation duration, hospital stay, cranial index, fronto-orbital retraction rate and complication rate between the two groups (all P>0.05). The forehead shape of the study group was more symmetrical than that of the control group, the difference was statistically significant ( P<0.05). Conclusion:The effect of frontal orbital reconstruction and distraction osteogenesis using 3D-RM in the treatment of Crouzon syndrome is positive, the head shape and orbit are significantly improved. There were no serious complications after operation. It can play a good auxiliary role in the operation.

Objective:To investigate the effect of intrafrontal-orbital distraction osteogenesis in the treatment for school-aged children with severe proptosis in Crouzon syndrome.Methods:We retrospectively analyzed the clinical data of school-aged children with Crouzon syndrome treated with intrafrontal orbital distraction osteotomy at the Children’s Hospital of Nanjing Medical University from January 2016 to February 2021, and counted and analyzed the clinical results, complications (cerebrospinal fluid leakage, infection, etc.), and surgical safety.Results:A total of 12 children, 9 males and 3 females, aged 6-12.1 years, with an average age of 7.1 years, were included; 4 were cranial, 6 were craniofacial, and 2 were other types. All were shown with obvious proptosis. The surgical procedure was successful. Follow-up ranged from 3 to 20 months, with a mean of 14 months. The postoperative distraction distance was (19.1±3.3) mm, ranging from 15.2 to 25.6 mm. And the mean cranial index (CI) before and after surgery were 97.5% and 80.0%, respectively, with good craniofacial appearance and better improvement of proptosis and posterior cranial flattening. There were no other complications but for one child with infection at the external site of the distraction rod.Conclusions:The effect of frontal-orbital distraction osteotomy in the treatment of Crouzon syndrome with severe proptosis in school-aged children was positive, with significant improvement in proptosis and low incidence of postoperative cerebrospinal fluid leakage and infection. This procedure is a good choice for older children with severe proptosis.

Objective:To investigate the effect of intrafrontal-orbital distraction osteogenesis in the treatment for school-aged children with severe proptosis in Crouzon syndrome.Methods:We retrospectively analyzed the clinical data of school-aged children with Crouzon syndrome treated with intrafrontal orbital distraction osteotomy at the Children’s Hospital of Nanjing Medical University from January 2016 to February 2021, and counted and analyzed the clinical results, complications (cerebrospinal fluid leakage, infection, etc.), and surgical safety.Results:A total of 12 children, 9 males and 3 females, aged 6-12.1 years, with an average age of 7.1 years, were included; 4 were cranial, 6 were craniofacial, and 2 were other types. All were shown with obvious proptosis. The surgical procedure was successful. Follow-up ranged from 3 to 20 months, with a mean of 14 months. The postoperative distraction distance was (19.1±3.3) mm, ranging from 15.2 to 25.6 mm. And the mean cranial index (CI) before and after surgery were 97.5% and 80.0%, respectively, with good craniofacial appearance and better improvement of proptosis and posterior cranial flattening. There were no other complications but for one child with infection at the external site of the distraction rod.Conclusions:The effect of frontal-orbital distraction osteotomy in the treatment of Crouzon syndrome with severe proptosis in school-aged children was positive, with significant improvement in proptosis and low incidence of postoperative cerebrospinal fluid leakage and infection. This procedure is a good choice for older children with severe proptosis.

Objective:To construct the pET30a-EgG1Y162-2 prokaryotic expression plasmid and induce the expression of EgG1Y162-2 protein, so as to provide a research basis for development of Echinococcus granulosus vaccine. Methods:Using Echinococcus granulosus cDNA as a template, the target gene of EgG1Y162-2 was synthesized by PCR, and after digestion with restriction enzymes EcoRⅠ and Hind Ⅲ, it was connected to the prokaryotic expression vector pET30a to construct the recombinant plasmid pET30a-EgG1Y162-2. The recombinant plasmid was transformed into competent cell BL21 (DE3) and induced by isopropyl β-D-thiogalactoside (IPTG) to express a large number of proteins. The recombinant protein was purified by affinity chromatography. The purification level was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression product was identified by Western blotting. Results:The recombinant plasmid pET30a-EgG1Y162-2 was successfully constructed. After inducting expression, the bacterial supernatant and the eluate were both at a relative molecular weight of about 15 × 10 3, and the protein antigen component eluted with 200 mmol/L imidazole was relatively pure. Western blotting results showed that the purified recombinant protein EgG1Y162-2 with His tag could be recognized by His monoclonal antibody. Conclusion:The pET30a-EgG1Y162-2 prokaryotic expression plasmid of Echinococcus granulosus is successfully constructed, and the recombinant protein of EgG1Y162-2 is induced to express, laying a foundation for further study on anti- Echinococcus granulosus vaccine.

Objective:To investigate clinical effects of the three-dimensional(3D) digital technique-assisted distraction osteogenesis in newborns with Pierre Robin sequence(PRS).Methods:The data of neonatal Pierre Robin sequence treated with 3D digital technique-assisted distraction osteogenesis in Children’s Hospital Affiliated to Nanjing Medical University from April 2017 to April 2020 was retrospectively analyzed. 3D digital technique was used to assist the pre-operative design and osteotomy navigation template was made with 3D printing technique to guide in the mandibular osteotomy. CT scans were obtained 3 months postoperatively to follow up the growth of mandible. The rates of recurrence, titanium nail falling, infection, secondary operation and nerve injury were statistically analyzed. The counting data were analyzed using chi-square test. The independent influencing factors of postoperative complications were analyzed by multiple logistic regression, and the difference was statistically significant when P<0.05. Results:Three hundred and forty-five patients were included in the 3D digital technique group consisting of 215 males and 130 females with the ages ranged 3-28 days and the weights 1.8-3.2 kg. Surgical results were satisfied and very close to the preoperative design. Craniofacial CT results showed normal growth of mandible 3 months postoperatively. Three hundred and seventy-seven patients were included in the controlled group consisting of 230 males and 147 females with the ages ranged 6-28 days and the weights 1.6-3.6 kg. The recurrence rate, extender titanium nail off rate, reoperation rate and risk of nerve damage in the 3D digital technique group were 0.29%(1/345), 0.29%(1/345), 0.58% (2/345)and 0.29%(1/345), respectively, while in the control group, the numbers were 1.86%(7/377), 2.92%(11/377), 2.65%(10/377), 2.92% (11/377), respectively, which were significant lower than those in the 3D group. The infection rates were 7.83%(27/345) in the 3D digital technique group and 7.69%(29/377) in the control group, and there was no significant difference between the two groups. The results of multi-logistic regression analysis suggested that the 3D digital technique group could be an independent factor to reduce postoperative complications ( P=0.001). Conclusions:In the newborns with Pierre Robin sequence, 3D digital technique-assisted distraction osteogenesiscan significantly improve the accuracy and success rate of the operation and reduce the complication rate as well.

Objective:To investigate clinical effects of the three-dimensional(3D) digital technique-assisted distraction osteogenesis in newborns with Pierre Robin sequence(PRS).Methods:The data of neonatal Pierre Robin sequence treated with 3D digital technique-assisted distraction osteogenesis in Children’s Hospital Affiliated to Nanjing Medical University from April 2017 to April 2020 was retrospectively analyzed. 3D digital technique was used to assist the pre-operative design and osteotomy navigation template was made with 3D printing technique to guide in the mandibular osteotomy. CT scans were obtained 3 months postoperatively to follow up the growth of mandible. The rates of recurrence, titanium nail falling, infection, secondary operation and nerve injury were statistically analyzed. The counting data were analyzed using chi-square test. The independent influencing factors of postoperative complications were analyzed by multiple logistic regression, and the difference was statistically significant when P<0.05. Results:Three hundred and forty-five patients were included in the 3D digital technique group consisting of 215 males and 130 females with the ages ranged 3-28 days and the weights 1.8-3.2 kg. Surgical results were satisfied and very close to the preoperative design. Craniofacial CT results showed normal growth of mandible 3 months postoperatively. Three hundred and seventy-seven patients were included in the controlled group consisting of 230 males and 147 females with the ages ranged 6-28 days and the weights 1.6-3.6 kg. The recurrence rate, extender titanium nail off rate, reoperation rate and risk of nerve damage in the 3D digital technique group were 0.29%(1/345), 0.29%(1/345), 0.58% (2/345)and 0.29%(1/345), respectively, while in the control group, the numbers were 1.86%(7/377), 2.92%(11/377), 2.65%(10/377), 2.92% (11/377), respectively, which were significant lower than those in the 3D group. The infection rates were 7.83%(27/345) in the 3D digital technique group and 7.69%(29/377) in the control group, and there was no significant difference between the two groups. The results of multi-logistic regression analysis suggested that the 3D digital technique group could be an independent factor to reduce postoperative complications ( P=0.001). Conclusions:In the newborns with Pierre Robin sequence, 3D digital technique-assisted distraction osteogenesiscan significantly improve the accuracy and success rate of the operation and reduce the complication rate as well.

Background and Objectives: Cranial sutures play a critical role in adjustment of skull development and brain growth. Premature fusion of cranial sutures leads to craniosynostosis. The aim of the current study was to culture and characterize human cranial suture mesenchymal cells in vitro. Methods: The residual skull tissues, containing synostosed or contralateral suture from three boys with right coronal suture synostosis, were used to isolate the suture mesenchymal cells. Then, flow cytometry and multilineage differentiation were performed to identify the typical mesenchymal stem cell (MSC) properties. Finally, we used quantitative real-time polymerase chain reaction (RT-PCR) to detect the mRNA expression of osteogenesis and stemness related genes. Results: After 3 to 5 days in culture, the cells migrated from the tissue explants and proliferated parallelly or spirally. These cells expressed typical MSC markers, CD73, CD90, CD105, and could give rises to osteocytes, adipocytes and chondrocytes. RT-PCR showed relatively higher levels of Runx2, osteocalcin and FGF2 in the fused suture MSCs than in the normal cells. However, BMP3, the only protein of BMP family that inhibits osteogenesis, reduced in synostosed suture derived cells. The expression of effector genes remaining cell stemness, including Bmi1, Gli1 and Axin2, decreased in the cells migrated from the affected cranial sutures. Conclusions The MSCs from prematurely occlusive sutures overexpressed osteogenic related genes and down-regulated stemness-related genes, which may further accelerate the osteogenic differentiation and suppress the self-renewal of stem cells leading to craniosynostosis.

Objective:To investigate the treatment of craniosynostosis in small infants with posterior vault distraction osteogenesis and the reasonable treatment for lambdoid sutures during operation.Methods:Seven cases with syndromic craniosynostosis who were admitted to our department from January 2017 to August 2019 were osteotomized by piezosurgery supraoccipital protuberance osteotomy in the posterior cranium, and two 2 cm distractors were placed in the lateral cranium. In 4 of 7 cases bilateral lambdiod sutures were fixed with titanium plate, while 3 of 7 were not fixed. Distraction was started from 5 days postoperatively, once a day on each side with a rate of 0.4-0.8 mm/d on each side for 10-15 days. The distractors were removed 6 months postoperatively. The titanium plates fixation in 4 cases were also removed at the same time.Results:The patients were followed up for 6 to 14 months, with an average of 12 months. The skull cavity and posterior cranial shape were observed. The posterior vault was distracted completely with lambdoid suture fixation in 4 cases. In the other 3 cases, the posterior skull was flattened. All skull deformities were significantly improved without death, skull necrosis, intracranial infection and other serious complications. The cranial index of both groups of lambdoid suture fixed and unfixed were reduced after the operation, and the fixed group was reduced by an average of 20.5%±5.8%, higher than the unfixed group(17.5%±5.1%), but the difference between the two groups was not statistically significant( P>0.05). The occipitofrontal circumference was increased after the operation in the two groups. The fixed group was increased by (2.7±0.4)cm on average, which was significantly higher than the unfixed group(1.7±0.1) cm. And the difference was statistically significant( t=3.789, P=0.013). Conclusions:Posterior vault distraction osteogenesis is a good method to treat the syndromic craniosynostosis. But bilateral lambdoidsuture needs to be fixed in infants. Titanium plate fixation could make the posterior cranium into a whole cranial plate for posterior vault distraction osteogenesis. And the posterior vault was obviously flat without lambdoid sutures fixation. Flatted occipital contour was improved significantly and the result of the fixed group was superior to the unfixed one. Bilateral fixation of lambdoid sutures is suitable for the infants, especially for severe syndromic craniosynostosis.

Objective:To investigate the treatment of craniosynostosis in small infants with posterior vault distraction osteogenesis and the reasonable treatment for lambdoid sutures during operation.Methods:Seven cases with syndromic craniosynostosis who were admitted to our department from January 2017 to August 2019 were osteotomized by piezosurgery supraoccipital protuberance osteotomy in the posterior cranium, and two 2 cm distractors were placed in the lateral cranium. In 4 of 7 cases bilateral lambdiod sutures were fixed with titanium plate, while 3 of 7 were not fixed. Distraction was started from 5 days postoperatively, once a day on each side with a rate of 0.4-0.8 mm/d on each side for 10-15 days. The distractors were removed 6 months postoperatively. The titanium plates fixation in 4 cases were also removed at the same time.Results:The patients were followed up for 6 to 14 months, with an average of 12 months. The skull cavity and posterior cranial shape were observed. The posterior vault was distracted completely with lambdoid suture fixation in 4 cases. In the other 3 cases, the posterior skull was flattened. All skull deformities were significantly improved without death, skull necrosis, intracranial infection and other serious complications. The cranial index of both groups of lambdoid suture fixed and unfixed were reduced after the operation, and the fixed group was reduced by an average of 20.5%±5.8%, higher than the unfixed group(17.5%±5.1%), but the difference between the two groups was not statistically significant( P>0.05). The occipitofrontal circumference was increased after the operation in the two groups. The fixed group was increased by (2.7±0.4)cm on average, which was significantly higher than the unfixed group(1.7±0.1) cm. And the difference was statistically significant( t=3.789, P=0.013). Conclusions:Posterior vault distraction osteogenesis is a good method to treat the syndromic craniosynostosis. But bilateral lambdoidsuture needs to be fixed in infants. Titanium plate fixation could make the posterior cranium into a whole cranial plate for posterior vault distraction osteogenesis. And the posterior vault was obviously flat without lambdoid sutures fixation. Flatted occipital contour was improved significantly and the result of the fixed group was superior to the unfixed one. Bilateral fixation of lambdoid sutures is suitable for the infants, especially for severe syndromic craniosynostosis.