Objective To investigate the expression of specific marker molecules in hair-inducing activity of long-term cultured human dermal papilla cells (HDPCs) in vitro.Methods After dissected and cultured the HDPCs in vitro,the cells of passages 1 to 8 were used for experiments.The growth appearances of HDPCs in different passages were observed under inverted microscope.To detect the expression of specific marker molecules of long-term cultured HDPCs,the alkaline phosphatase (ALP) activity of the HDPCs was examined,and the specific genes ALP and insulin-like growth factor-1 (IGF-1) expression levels of HDPCs were determined by real-time quantitative PCR.Results After long-term cultured in vitro,the ALP and IGF-1 expression levels of HDPCs gradually decreased in different passages,as well as the display of the aggregated and cartouche growth.The ALP and IGF-1 expression levels of HDPCs in passage 1 was the highest,they were almost about 6.8-fold and 3.5-fold higher than the HDPCs in passage 8.The ALP staining of the HDPCs in passage 1 and passage 2 were evident,but the cells' ALP staining gradually became much weaker than the cells in the previous passages after the long-term cultured in vitro.Conclusions The expression levels of specific marker molecules ALP and IGF-1 of the HDPCs decrease gradually after long-term cultured in vitro,and the higher passage HDPCs lost the special aggregated and cartouche growth appearance,and hence lead to the loss of hair-inducing activity of HDPCs.

Objective To establish a rat skin organ culture model to study the initiation of hair follicle morphogenesis, and the dynamic expression of Wnt10b/β-catenin in the developing hair follicle. Methods The dorsal skins of SD rat at embryos 14-17 (E14-E17) were cultured on a gelatin sponge-supported tissue culture system at the air/liquid interface of DMEM with 10 % fetal bovine serum (FBS) for 3-6 days, some of which removed from El5 were cultured in DMEM with FBS of different concentrations. HE staining and fluoroimmunoassay were adopted. Results The model we established allowed skin tissues isolated from E14-E15 to develop in a manner that was histologically and temporarily similar to the process in vivo. However, the developing hair follicle ceased to continue when their morphogenetic process reached the forth stage, and the concentration of FBS did not show any significant effect on the development of hair follicle. Expression of Wnt10b/β-catenin was induced in culture, as it also occurred in vivo,but grew weak till it disappeared in culture for 6 days, which was accompanied by development halt of hair follicle. Conclusions A rat skin organ culture model is established in which the morphogenesis of hair follicle takes place in a manner similar to in vivo, and Wnt10b/β-catenin probably has a close connection with the early morphogenesis of hair follicle.

Objective To evaluate the relationship between birth weight and nonalcoholic fatty liver diseases (NAFLD) in Chinese primary school students. Methods A cross-section study was conducted in ifve elementary schools in Gao Hang Town, Shanghai and 2163 students were enrolled in the study (1120 boys/1043 girls). Height, body weight, waist circumference and per-cent of body fat (bioelectrical impedance analysis) were measured by professional nutritionist after training. Birth weight, feeding pattern, height and body weight of parents were obtained by a self-completed questionnaire. NAFLD was diagnosed by ultra-sound. The risk factors of NAFLD were analyzed. Results The prevalence of NAFLD in the study population was 8.9%. The prevalence of NAFLD was signiifcantly higher in boys than that in girls (12.5%vs 5.0%, P<0.01). Logistic regression showed that sex (OR=1.97, 95%CI:1.21-3.21) and percent of body fat (OR=1.12, 95%CI:1.07-1.17) were the risk factors of NAFLD, and normal BMI was the protective factor of NAFLD (OR=0.09, 95%CI:0.04-0.19) in the study population. Conclusions The pre-valence of NAFLD is higher in boys than that in girls. Also overweight, and high percent of body fat are risk factors of NAFLD in children.

BACKGROUND: Mandibular distraction osteogenesis (MDO) is a kind of endogenous tissue engineering technology that lengthens the jaw and opens airway so that a patient can breathe safely and comfortably on his or her own. Endothelial progenitor cells (EPCs) are crucial for MDO-related angiogenesis. Moreover, emerging evidence suggests that heat shock protein 20 (Hsp20) modulates angiogenesis under hypoxic conditions. However, the specific role of Hsp20 in EPCs, in the context of MDO, is not yet known. The aim of this study was to explore the expression of Hsp20 during MDO and the effects of Hsp20 on EPCs under hypoxia. METHODS: Mandibular distraction osteogenesis and mandibular bone defect (MBD) canine model were established. The expression of CD34, CD133, HIF-1α, and Hsp20 in callus was detected by immunofluorescence on day 14 after surgery. Canine bone marrow EPCs were cultured, with or without optimal cobalt chloride (CoCl2 ) concentration. Hypoxic effects, caused by CoCl2 , were evaluated by means of the cell cycle, cell apoptosis, transwell cell migration, and tube formation assays. The Hsp20/KDR/PI3K/Akt expression levels were evaluated via immunofluorescence, RT-qPCR, and western blot. Next, EPCs were incorporated with either Hsp20-overexpression or Hsp20-siRNA lentivirus. The resulting effects were evaluated as described above. RESULTS: CD34, CD133, HIF-1α, and Hsp20 were displayed more positive in the callus of MDO compared with MBD. In addition, hypoxic conditions, generated by 0.1 mM CoCl2 , in canine EPCs, accelerated cell proliferation, migration, tube formation, and Hsp20 expression. Hsp20 overexpression in EPCs significantly stimulated cell proliferation, migration, and tube formation, whereas Hsp20 inhibition produced the opposite effect. Additionally, the molecular mechanism was partly dependent on the KDR/PI3K/Akt pathway. CONCLUSION In summary, herein, we present a novel mechanism of Hsp20-mediated regulation of canine EPCs via Akt activation in a hypoxic microenvironment.

Objective:To establish silicone cartilage models of donor-sites for the microtia patients by using digital technology, and to explore the application of surgical simulation in auricular reconstruction.Methods:From June 2018 to October 2019, 19 congenital microtia patients underwent thoracic CT scans and following three-dimensional costal cartilage imaging with Mimics software at the Nanfang Hospital, Southern Medical University. Among these patients, 16 were males and 3 were females. The mean age of patients was 16 years (range 8 to 35 years). Silicon cartilage models were produced by 3D printing and used for surgical planning and preoperative simulation in ear framework fabrication. Cartilaginous framework was sculptured according to the simulation during operation. Patients were followed up for a minimum of six months to evaluate the size, outline, height and auriculocephalic angle of the reconstructed ear. The satisfactory outcomes of the patients were scored according to a 5-point Likert scale.Results:All the patients received the surgical simulation and sculpture training with silicone cartilage models before operation. Auricular reconstruction was completed successfully according to the simulation. The duration of sculpture was shortened to 1-1.5 hours. There were no serious complications, such as hematoma, inflammation, skin necrosis and framework exposure. The contour of reconstructed ear was natural and clear over a 6 months follow-up, and all the patients were satisfied with their surgical outcomes.Conclusions:With the application of digital technology and silicone cartilage models by 3D printing to the surgical planning and training in microtia patients, patient-specific framework is fabricated with precisely assembling, which not only shortens the operation time, but also provides the unexperienced surgeons with a safe and effective training of ear framework fabrication.

Objective:To investigate the application of three-dimensional digital technique in customized ear framework fabrication for auricular reconstruction.Methods:From July 2018 to October 2019, the patients with microtia who underwent ear reconstruction in the Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University were enrolled. Each patient with unilateral microtia underwent auricular CT scan and preoperative analysis and ear framework design were carried out with Mimics software 18.0. The two-dimension(2D) ear films and three-dimension(3D) silicon models were produced by 1∶1 2D printing and 3D printing, respectively. Microtia reconstruction was performed according to the guide of the models, patients were followed up over a six-month period to evaluate the size, outline, height and auriculocephalic angle of the reconstructed ear. The satisfactory outcomes of the patients were scored according to a 5-point Likert scale.Results:A total of 15 patients were included in this study, including 11 males and 4 females, aged 8-27 years, with an average of 15.5 years old. All the 15 patients completed the surgical planning and ear reconstruction successfully, without major complications, such as hematomas, inflammation, skin necrosis and framework exposure. The costal cartilage frameworks were very similar to the printed 3D models in size and contour. Comparison between the two sides was made at six months postoperatively. The reconstructed ear was much the same as that of contralateral side, and all patients were satisfied with their reconstructed ear outcomes with average score of 4.4.Conclusions:With the application of digital technique for pre-surgical planning in microtia reconstruction patients, ear templates were produced from 2D to 3D, and the correction of microtia was changed from standard auricular reconstruction to personalized auricular reconstruction, with a great improvement of the precision in ear framework fabrication.

Objective:To introduce a reliable and easy method of philtrum crest and philtrum repairing in secondary repair of cleft lip.Methods:Lift the upper lip flap after removing the scar on upper lip. Then the superficial orbicularis oris muscle was carefully separated, which formed the abnormal muscular eminence in the affected side after the first cleft lip repair. The pedicle of the orbicularis oris muscle the reconstructing philtrum crest. The lingual orbicularis oris muscle flap was lifted and curled from outside. The superficialorbicularis oris muscle was cut in philtrum longitudinally and turned outward. The two evertedorbicularisoris muscle flaps were sutured on both sides and fixed them on the reconstructing philtrum crest. The exposed deep orbicularis oris muscle in philtrum was sutured with the upper dermis, which formed the prominentphiltrum crest and deeper philtrum.Results:We reviewed the hospital records of patients from July 2008 to November 2021. In total, 201 patients were included in this study. All of these patients underwent the reconstruction of philtrum crest and philtrum with orbicularis oris muscle intertangling and philtrum plasty in secondary cleft lip repairing. During the treatment, the patients had no complications such as infection, hematoma and flap necrosis.198 patients were observed from 1 months to 36 months (7.8 months on average). All of them improved in different degrees in early postoperative period. Most reconstructed philtrum crest was raised and basically symmetrical with the contralateral side in shape. The philtrum was clear and the scar was not obvious. The reconstructed philtrum crest would decrease with time. 91.7% (90/98) patients were satisfied with the postoperative effects after three months.Conclusions:The method of reconstruction of philtrum crest and philtrum with orbicularis oris muscle intertangling and philtrum plasty in secondary cleft lip repairing is safe and effective. The long term postoperative effect is stable with high satisfaction as well. As a result, this method can be used as the philtrum crest and philtrum correction scheme for secondary cleft lip repairing.

Objective:To investigate the application of three-dimensional digital technique in customized ear framework fabrication for auricular reconstruction.Methods:From July 2018 to October 2019, the patients with microtia who underwent ear reconstruction in the Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University were enrolled. Each patient with unilateral microtia underwent auricular CT scan and preoperative analysis and ear framework design were carried out with Mimics software 18.0. The two-dimension(2D) ear films and three-dimension(3D) silicon models were produced by 1∶1 2D printing and 3D printing, respectively. Microtia reconstruction was performed according to the guide of the models, patients were followed up over a six-month period to evaluate the size, outline, height and auriculocephalic angle of the reconstructed ear. The satisfactory outcomes of the patients were scored according to a 5-point Likert scale.Results:A total of 15 patients were included in this study, including 11 males and 4 females, aged 8-27 years, with an average of 15.5 years old. All the 15 patients completed the surgical planning and ear reconstruction successfully, without major complications, such as hematomas, inflammation, skin necrosis and framework exposure. The costal cartilage frameworks were very similar to the printed 3D models in size and contour. Comparison between the two sides was made at six months postoperatively. The reconstructed ear was much the same as that of contralateral side, and all patients were satisfied with their reconstructed ear outcomes with average score of 4.4.Conclusions:With the application of digital technique for pre-surgical planning in microtia reconstruction patients, ear templates were produced from 2D to 3D, and the correction of microtia was changed from standard auricular reconstruction to personalized auricular reconstruction, with a great improvement of the precision in ear framework fabrication.

Objective To explore the feasibility of percutaneous puncture for ventricular septal defect(VSD).Methods From March 2015,we did percutaneous punctured transcatheter device closure of VSD for 22 patients.The patients included 15 males and 7 females with age from 2 years 6 months to 11 years 7 months, weight from 13.5 kg to 44.0 kg.Among 22 pa-tients,6 were diagnosed with residual shunt after VSD repair, with size of residual shunt from 4.8 mm to 7.0 mm.Residual shunts are perimembranous, conoventricular and intracristal.The other 16 patients were diagnosed with isolated VSD,with de-fect size from 3.5 to 5.1 mm.For all patients,we punctured directly through the right ventricular surface into the right ventri-cle in the 3th or 4th intercostal space of the left sternal border.Next, we succeeded to extract the guide wire after inserting a conveyor tube for 21 patients.Subsequently,7-10 mm VSD closure device were delivered via the conveyor tube.Only one patient was changed to perform occlusion of VSD through chest small incision due to improper selection of punctured position. Results A total of 21 closure devices were placed for 21 patients.18 of them are equilateral device,and 3 are decentered de-vice,with size from 5 mm to 10 mm.There were small amount of pericardial effusion in 4 patients using TEE examination,and no future treat was performed after observation.For one-year follow-up, all patients have recovered very well.Conclusion Percutaneous punctured transcatheter device closure fits for treatment for children with residual shunt after VSD repair and isola-ted VSD.It has a good recent result.