Isolation and Characterization of Human Suture Mesenchymal Stem Cells In Vitro

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.

Management of nasal deformity with unicoronal craniosynostosis using the nasal bones were wedge removed / 中华整形外科杂志

Objective@#To investigate the treatment of nasal deformity in patients with unicoronal craniosynostosis.@*Methods@#In patients over 6 months old, the nasal bones were wedge-removed without fixation. The management of all patients with unicoronal craniosynostosis was distraction osteogenesis of pedicled unilateral frontal bone flap.@*Results@#Postoperative extended distance of the frontal bone was 28—41 mm (mean, 35.4 mm). After extension, three-dimensional reconstruction of cranial CT was carried out, which showed that CVAI was 0.8—1.2 (mean, 0.98), tending to normal. After discharge, dynamic cranial braces were put on for 1 year. Postoperatively, the children were followed up for 8—36 months (mean, 28 months). The shape and nasal deformity of all children were improved compared with those before surgery.@*Conclusions@#Nasal wedge resection should be used to correct nasal deformity in children over 6 months with unicoronal craniosynostosis.

Treatment of syndromic craniosynostosis with distraction osteogenesis / 中华整形外科杂志

Objective@#To explore the effect of distraction osteogenesis in the treatment of syndromic craniosynostosis.@*Methods@#The clinical data of 6 children with syndromic craniosynostosis from January 2014 to September 2018 were retrospectively analyzed. There were 5 males and 1 female, aged from 1 month and 21 days to 6 years and 1 month, with an average age of 30 months. There were 3 Crouzon syndrome, 1 Pfeiffer syndrome, 1 Vogt syndrome (ACS Ⅱ) and 1 Clove leaf skull syndrome. The distraction osteogenesis apparatus was used in this procedure. The distraction osteogenesis was prolonged twice a day, 0.4 mm each time, and the prolongation was stopped when the skull shape was significantly improved. Three-dimensional CT scans of the skull were reviewed after 6 months, suggesting that distraction osteogenesis was good, then the lengthener was removed. Complications were recorded, and extended distances were measured. Cranial indices before and after operation were compared to evaluate the efficacy, safety and feasibility of the operation.@*Results@#The procedure of operation and traction was successful in all 6 children. There were no complications such as cranial spinal fluid (CSF) leakage or intracranial infection occurred. The increased distance was (19.1±3.3) mm, ranging from 15.2 to 25.6 mm. The preoperative cranial index was 89.6 ±7.3, while the postoperative cranial index was 74.2 ±3.6. All patients were followed up from 3 to 20 months, with the average of 14 months. The posterior cranial flatness was improved, and the patients were satisfied with the surgical results.@*Conclusions@#Distraction osteogenesis is effective and reliable in treating the premature fusion of cranial suture, and it produces excellent result with low rate of CSF leak and infection.

Effects of Bmi1 on proliferation of cranial suture mesenchymal cells in mice / 中华整形外科杂志

Objective@#To explore the effects of Bmi1 on proliferation of mouse cranial suture mesenchymal cells.@*Methods@#Primary posterior frontal and sagittal suture derived cells were isolated from the 2-5 d old C57BL/6 suckling mice (n=6) of the same brood and cultured. Flow cytometry and multilineage differentiation assay were performed to identify the mesenchymal stem cells (MSCs) characteristics of the 2 kinds of cranial suture-derived cells. The mRNA expression of stem cell related genes, Bmi1, Twist1, Gli1 and Axin2 were detected by real-time quantitative polymerase chain reaction (RT-PCR). Then, the proliferation and downstream protein expression were analyzed after down-regulation of Bmi1 in the sagittal suture derived MSCs by transfecting Bmi1 siRNA. The t test was used to compare the mean between two groups. Statistical significance was set at P<0.05.@*Results@#The mouse cranial suture derived cells were successfully cultured in vitro. These cells expressed typical MSCs markers, CD44, CD90, CD73, except for CD34. These cells had osteogenic, adipogenic and chondrogenic differentiation potency. RT-PCR results showed that the mRNA expressions of Bmi1 (0.006 30±0.000 58 vs 0.002 60±0.000 34, t=5.430, P=0.005 6), Twist1(0.000 31±0.000 04 vs 0.000 15±0.000 02, t=3.343, P=0.028 8), Axin2(0.000 33±0.000 03 vs 0.000 17±0.000 05, t=3.067, P=0.037 4) and Gli1 (0.001 10±0.000 13 vs 0.000 60±0.000 33, t=3.956, P=0.016 7) were significantly decreased in the posterior frontal suture MSCs compared with those in sagittal suture derived cells. Among them, Bmi1 has the largest decline. After down-regulation of Bmi1 in sagittal suture MSCs, the protein expression level of Ink4a was significantly up-regulated compared with the control group, and the cell proliferation ability was significantly decreased.@*Conclusions@#Inhibition of Bmi1 expression can up-regulate the expression of Ink4a protein and decrease the proliferation ability of suture MSCs, which may lead to craniosynostosis.

Treatment of pediatric maxillofacial fractures using distraction osteogenesis / 中华整形外科杂志

Objective@#To investigate the clinical outcome of distraction osteogenesis in the treatment of pediatric maxillofacial fractures.@*Methods@#From November 2017 to November 2018, 6 cases of maxillofacial fractures were treated, including 3 cases of maxillary fracture and 3 cases of mandibular fracture. All of them were associated with facial asymmetry of different severity, disordered occlusion and displacement of fracture segments. The distraction osteogenesis device was used to fix the fracture during the operation. The distraction was initiated after 1 day latency period, and proceeded at approximately 0.5 mm each time, twice a day, until the normal occlusal relationship was achieved. Distractor was maintained for 1 month after distraction, and then removed. The degree of fracture healing, the mouth opening and the occlusal status at the fracture site were evaluated during follow-up, and the corresponding curative effect was observed and evaluated.@*Results@#All fractures were healed well, without infection, dislocation, or pseudo-joint formation after 1 month. The distraction duration is 15-20 days and the distraction distance is 15-20 mm. At the end, the occlusal relationship was normal, and the mouth opening was not limited. The follow-up period was from 1 month to 1 year. The maxillofacial appearance of the children was normal, and the maxillofacial development was not significantly affected.@*Conclusions@#Distraction osteogenesis is an effective and controllable method for child maxillofacial fracture. It can be used as a supplement to the conventional treatment of maxillary and mandibular fracture in children.

Multiple osteotomies with combined distraction osteogenesis for non-syndromic scaphocephaly / 中华整形外科杂志

Objective@#To evaluate the clinical effect of multiple osteotomies combined with distraction osteogenesis in the treatment of non-syndromic scaphocephaly infants and young children.@*Methods@#Clinical records of 8 non-syndromic scaphocephaly patients, during January 2017 to October 2018, from the Department of Burn and Plastic Surgery, Children′s Hospital of Nanjing Medical University, were retrospectively analyzed. There were 3 males and 5 females, aged from 3 to 28 months, with an average of 13 months. Among them, 4 cases were typical scaphocephaly, 2 cases were anterior scaphocephaly and 2 cases were posterior scaphocephaly. According to the subtypes of scaphocephaly, different osteotomy methods and distraction directions were designed. The cranial vault was osteotomized by piezosurgery osteotomy in a multiple way. The dura did not separate from cranial flap. The internal distractors were installed subsequently. After a mean latency period of 5 days, the devices were activated at a rate of 0.4-0.6 mm/d. Distractor was extended twice a day. After 10-15 days activation period and 6 months consolidation, all the distractors were removed. Three-dimensional CT scans were performed before and after operation to measure the anteroposterior and bitemporal diameters, so as to calculate the cranial index.@*Results@#Follow-up ranged from 6 to 14 months (average, 10 months). The average anteroposterior length of 8 children was(158.2±12.9) mm before operation, and (145.5±14.2) mm after operation, which was shortened(12.6±3.4) mm. Bitemporal diameters was(99.6±8.6) mm preoperatively and(113.9±7.5) mm postoperatively, which was widened (14.2±2.8) mm. Preoperative and postoperative cranial index was 63.2 and 78.3, respectively. The scaphocephalic head shape was significantly improved in all patients. There was no serious complications such as skull necrosis or intracranial infection.@*Conclusions@#Different types of multiple osteotomy combined with distraction osteogenesis are useful to correct various scaphocephaly of infants and children, by improving skull shape and cranial index.

Mandibular distraction osteogenesis to treat Braddock-Carey airway obstruction / 中华整形外科杂志

Objective@#To explore the feasibility of mandibular distraction for the treatment of airway obstruction in neonates with Braddock-Carey syndrome.@*Methods@#From August 2007 to November 2017, 6 neonates with Braddock-Carey syndrome were treated in our center. All patients experienced the combined thrombopoietic treatment before operation. Bilateral oblique mandibular osteotomy was performed by extraoral approaches and the distraction was initiated on postoperative day 3 at a rate of 1.2 mm/d. It usually took around 2 weeks to get required length of distraction. Distractor was removed after 3-months fixation at second stage.@*Results@#The distraction distance of 6 cases was from 12 to 20 mm. The CT scan showed a widening gap of the airway by 6-10 mm, and the average width was around 7 mm. 5 patients had no bleeding and infection after the operation, while one patient experienced bleeding for 4 times treated by regional compression. All neonates felt painful during distraction, and got relieved after reduction of the frequency and pace. With mandibular advancement, all patients′ face type improved, while paroxysmal bruising, inspiratory dyspnea and difficulty in breastfeeding disappeared. After 6 months of follow-up, all patients′ weight and face type were basically the same as normal children.@*Conclusions@#With the treatment of thrombocytopenia before operation, complete hematischesis during the operation, and postoperative treatment of thrombocytopenia, mandibular distraction osteogenesis was a feasible and safe management of upper airway obstruction in neonates with Braddock-Carey syndrome.

Neonatal RICH associated with thrombocytopenia and coagulopathy: a case report and literature review / 中华整形外科杂志

We treated a neonate who was diagnosed as rapidly involuting congenital hemangioma (RICH). With a review on the clinical manifestation, differential diagnosis, and treatment of RICH that associated with thrombocytopenia(TP)and coagulopathy(CP)and received platelet transfusions and embolization therapy including absolute ethanol and polidocanol in our hospital in March of 2015. The platelets and coagulation function soon returned to normal, the tumor involuted significantly. Surgical excision was proceeded subsequently. The platelets returned to normal level after a one-time platelet transfusion, meanwhile, multiple reexaminations of blood coagulation function were normal. Postoperatively, incision wound healed well. There was no recurrence, and the functional recovery of upper limb was satisfactory. RICH is a rare type of vascular tumor which may present with TP and CP similar to KHE-KMP in the neonatal period. More attention should be addressed to identify these two diseases, because the treatment and prognosis of which have significant differences. Consumption of coagulation factors with milder platelet decrease may also be a complication of slow flow venous or venolymphatic malformations.

Treating unilateral coronal synostosis in infant: internal forehead distraction and induced osteogenesis of supraorbital margin of frontal bone / 中华整形外科杂志

Objective@#To investigate a new method to treat unilateral coronal synostosis.@*Methods@#2 cm-wide osteotomy was performed over the fused unilateral coronal suture.Oblique osteotomy was performed along the supraorbital rim to do a frontal suture of the glabella (patients less than one year old) or on the opposite side of the supraorbital rim (patients over one year old) after performing a suturectomy of the effected coronal suture. Two internal distraction devices were subsequently placed across the osteotomized, fused coronal suture. At the top of the unfused metopic suture, bring two holes and fixing with 4-0 absorbable suture.Finally, the cranium pieces were divided into 2 pieces and placed in the middle of the frontal bone using biological glue or titanium screws. Five days after the operation, a 0.6 mm distraction was performed twice per day. The distraction was removed 6 months after distraction reaching 30—60 mm.@*Results@#Internal distraction osteogenesis with supraorbital oblique osteotomy was performed in eleven patients suffering from unilateral coronal synostosis from 2014.1 to 2017.1. All the patients had no abnormalities in nerve system, such as cephalomeningitis/ fistaul of cerebrospinal fluid/ epidural hematoma and so on.The distractor was not exposed. The width of extension ranged from 25—39 mm. One patient had infection in the rods around the distraction during the period of fixed, but was cured with antibiotic treatment and got an ideal extension eventually. The rest of 10 patients obtained the expected result . During a mean follow-up period of 12 months (5 to 26 months), all patients were presented with satisfactory cosmetic and functional result . CVAI were close to normal.@*Conclusions@#Treating unilateral coronal synostosis by internal distraction osteogenesis with supraorbital oblique osteotomy reduces the necessity offrontal and supraorbital osteotomy, and the exposure of the base of anterior cranial fossa, renders a safe and effective approach.

Three-dimensional Z-shaped osteotomy with cranium distraction osteogenesis for unilateral lambdoid synostosis / 中华整形外科杂志

Objective@#To evaluate the effect of three-dimensional Z-shaped osteotomy with cranium distraction osteogenesis for unilateral lambdoid synostosis.@*Methods@#Imaging the skull as a cuboid, we designed the affected lambdoid suture as the middle line of letter Z, the osteotomy line paralleling to the sagittal suture or extending downward posterior cranial fossa as the two horizontal lines of Z to obtain the three-dimensional Z-shaped osteotomy lines. Two or three distraction devices were installed after removing the cranial bone at the premature fused suture with a width of 2.0-2.5 cm. Since the 5th day after operation, distraction was performed at the rate of 0.6 mm each day, twice a day, until the distraction distance reached 2.0-4.5 cm. Finally, we removed the distractors after fixation for three months.@*Results@#Eleven cases of unilateral lambdoid synostosis were treated successfully. No infection or bleeding happened except for one case with distractor entrance scalp infection. No complications, including the fixed screws displacement, penetrating the cranium and the dura mater or distraction devices retracting, occurred. During a mean follow-up period of 24 months (5 to 36 months), all patients were satisfied with the cosmetic and functional result.@*Conclusions@#Three-dimensional Z-shaped osteotomy with cranium distraction osteogenesis can not only extend upward to raise the cranial height, but also lengthen backward to singularize the occipital carina, forming a well-pleasing occipitalia. Z shape stabilizes the whole plate, making the separated cranial bones not easy to retract. Therefore, using this procedure is effective and mini-invasive, and especially suitable for young infant.

Management for lymphatic malformation in infants: a single center experience / 中华整形外科杂志

Objective@#To assess the therapeutic effect of management for lymphatic malformation(LM)in infants.@*Methods@#This retrospective study recruited clinical data of 996 patients with LM from June 2004 to July 2015 in our center. All patients were diagnosed as LM after ultrasound, CT or MR scan. All patients were divided into Group 1 (427 patients, treated by endoscopic LM partial resection combined with cautery and postoperative intratumoral negative pressure and absolute ethyl alcohol), Group 2(239 patients, treated by combined pinyangmycin and dexamethasone injection), Group 3 (330 patients, treated by surgical resection only). The clinical effects were observed in three groups, and therapeutic effect differences in gender, age, maximum diameter, location, range, histological typing, lymph property and treatments were analysed in cervicofacial LM.@*Results@#Group 1: 333 patients were cured (78.0%), Group 2: 165 patients were cured(69.0%), Group 3: 238 patients were cured (72.1% ). The difference in cure rate between Group 1 and Group 2 or between Group 1 and Group 3 was significant(P<0.05). The number of patients with ≥2 treatments in Group 3 was significantly less than that in other two groups. The cure rate of LM in cervicofacial area was significantly lower than that in other parts of body (P<0.05). In cervicofacial LM patients, the therapeutic effect differences in maximum diameter, range, histological typing, lymph property were statistically significant (P<0.05). At the multivariable logistic regression analysis, LM range as well as histological typing were independent factors influencing the therapeutic effect (P<0.05).@*Conclusions@#Both treatment of endoscopic LM partial resection combined with cautery and postoperative intratumoral negative pressure and absolute ethyl alcohol, and surgical resection have good therapeutic effect on LM in infants. LM range as well as histological typing are important factors independently influencing the therapeutic effect of cervicofacial LM in infants.

Management of congenital median perineal cleft in children: a report of 7 cases / 中华整形外科杂志

<p><b>OBJECTIVE</b>To discuss the treatment and diagnosis of congenital median perineal cleft in children.</p><p><b>METHODS</b>Between January 2009 to February 2013, 7 cases were diagnosed as congenital median perineal cleft according to the symptoms. Among them, 4 cases underwent surgery to correct cleft with double triangular perineal flaps. The other 3 cases with minor cleft did not receive surgery management.</p><p><b>RESULTS</b>There is an median cleft from the perineum to the anus with mucosa on the cleft surface. Primary healing was achieved in all the four patients with satisfactory appearance. The patients were followed up for 1-4 years with almost normal perineal appearance.</p><p><b>CONCLUSIONS</b>Congenital perineal median cleft can be diagnosed according to the symptoms. Double triangular perineal flaps can be effectively correct the cleft to attain normal perineal appearance.</p>

Orthogonal Design of the Prescription of Selegiline Transdermal Patches / 中国医科大学学报

Objective To determine the prescription of selegiline transdermal patches.Methods The selegiline transdermal patches were prepared and the ingredient amount of selegiline transdermal patches was determined according to the orthogonal design.Results The best prescription of selegiline transdermal patches：3%selegiline,3%Azone,15%RS100,60%PSA.Conclusion Selegiline is suitable for the transdermal therapeutic systems.Orthogonal design of prescription was suitable to study selegiline transdermal patches.

Safety analysis of living related live donors in kidney transplantation / 中华泌尿外科杂志

Objective To analyze the clinical outcomes and evaluate the safety of living related live donors in kidney transplantation.Methods One hundre and thirty-two cases of living related donors were studied retrospectively for psychological and physiological parameters. The parameters including life quality, urinalysis, serum biochemistry tests, glomerular filtration rate (GFR) and endogenous creatinine clearance rate (CCr) were analyzed.Results There was no significant difference between living related donors and normal people in the aspect of life quality. In living related donors,SCr was (78.33±15.94)μmol/L before operation and was (108.49±19.88)μmol/L at 7 days postoperation, P＜0.05. SCr was (112.47±20. 38)μmol/L at 6 months post-operation. There was no significant difference in SCr levels between 7 days and 6 months post-operation (P=0. 109). CCr was (95.80±20.92)ml/min in pre-operation and was (57.36±14. 92)ml/min at 7 days post-operation,P＜0.05. CCr was (65. 49±8. 25) ml/min at 6 months post-operation. There was no significant difference in CCr between 7 days and 6 months post-operation.The pre-operative total GFR was(74.08±18.51)ml/min. Of which, the right kidney GFR was (38.43±10.33)ml/min. The residual right kidney GFR was (56. 49±13. 01 ) ml/min 6 months after operation, which decreased 17. 59 ml/min (23.8%) compared with pre-operative total GFR (P＜0.05) and increased 18. 06 ml/min (47.0%) compared to the pre-operative right kidney GFR. Surgical complications included 1 case of splenectomy, 1 case of descending colon rupture and 5 cases of wound fat liquefactions.Conclusion Pre-operatively systemic psychological and physiological evaluation on living related donors, detailed consent consultation, standardized operating techniques, careful perioperative management and strict follow-up can improve the safety of living related donors.

Analysis on spouse donor renal transplantation / 中华泌尿外科杂志

Objective To analyze clinical outcomes of the spouse donor kidney transplantation for evaluating the security.Methods Sixty-three cases of living-renal transplantation were divided into two groups, spouse donor group (12 cases) and the other related donor group (53 cases). Twelve cases of spouse-donor(SD)renal transplantation were summarized, that were compared with the nospouse transplantation cases of the other living-related renal transplantation in the same period, which were similar in basic conditions and in immunosuppressant scheme. The observational parameters included average hospitalization time, rate of acute renal necrosis, acute rejection incidence within 1 year, serum creatinine at 7 d, 30 d and 1 year after transplantation.Results The recipient age of the 2 groups was (39±3)years and (37±3)years(P=0.05), dialysis time was (4.7±3.2)months and (4.4±2.9)months(P=0.78), the average hospitalization time was (20.9±8.3)d and (23.0±7.8)d(P=0.41). There was no significant difference between the spouse donor group and the no spouse related donor group. The acute rejection incidence within 1 year was 33.3%(4/12) in spouse group and 3. 9%(2/51) in the other related group, there was significant difference between the 2 groups(P＜0.05). The rate of acute renal necrosis was 16.7%(2/12) in spouse group and 3.9%(2/15)in no spouse related group, there was no significant different between the 2 groups(P＞0.05).SCr was (206.47±47.22)μmol/L and (163. 75±25.91)μmol/L in spouse group at post-operation 7 d and 30 d, and was (142.79±89.42)μmol/L and (119. 99±15.03)μmol/L in no spouse group. There was significant difference between the groups(P=0.02, P=0.00). One year after operation, SCr was (133. 40±6. 11)μmol/L in spouse group and (121. 00±34.12)μmol/L in no spouse group,there was no significant difference between the 2 groups (P=0.25).Conclusions Preoperative comprehensive assessment of the spouse donors and recipients renal transplantation is the guarantee for the success.Short-term outcomes of spouse donor renal transplantation is ideal, but rate of acute rejection within 1 year is higher than that of the other living-relative donor kidney transplantation, which dose not influence the long-time survival of spouse recipients.

Laparoscopic splenectomy by dissecting in situ secondary braches of splenic pedicle for idiopathic thrombocytopenic purpura:report of 41 patients / 第三军医大学学报

Objective To evaluate the feasibility and clinical efficacy of amputation in situ secondary braches of splenic pedicle for laparoscopic splenectomy(LS)in patients of idiopathic thrombocytopenic purpura(ITP).Methods LS was performed in 41 ITP patients hospitalized in our department from January 2007 to November 2008 by dissecting secondary braches of splenic pedicle.The serosa on the spleen pedicle was opened by using a harmonic scalpel.Then secondary structures of the splenic arteries and veins,one by one,were disconnected from downward to upward,and then double ligated with the Hem-O-lok clips.The ligated section was cut off with the harmonic scalpe1,and then the spleen was resected.Results LS was successfully completed with no conversion to open surgery in all the 41 cases.The operative time was 75 to 180 min,and the estimated intra-operative blood loss was 50 to 800 ml.There were 3 cases of vice spleen and the vice spleen removed.Two cases were converted to traditional LS with Endo-GIA.The period of hospitalization was 4 to 9 d after operation.There was no severe postoperative complication.Platelet counts began to increase significantly after operation,reached to the peak in 5 to 7 d after operation.During the follow-up of 3 to 20 months to all 41 ITP patients,the total effective rate was 85.4%(35/41,including 22 cases of complete response and 13 cases of partial response).Conclusion LS with amputation in situ secondary braches of splenic pedicle is a safe,effective,minimally invasive and low complication in treatment of ITP.It is worth be recommended because of its lower costs in comparison with common LS.