Objective To investigate the expression of BRAF-activated long non-coding RNA (BANCR) in colorectal cancer,and the influence of BANCR on the biological function of HCT116 cells.Methods Fifty-six samples of colorectal cancer specimen (including the cancer tissues and precancerous tissues) were obtained at the First Affiliated Hospital of Nanjing Medical University from March 2012 to June 2013.The expressions of BANCR in all the specimens were detected by qRT-PCR (28 cases in the BANCR-high expression group and 28 cases in the BANCR-low expression group).The relationship between the expressions of BANCR and the clinicopathological factors of colorectal cancer was analyzed.The HCT116 cells were divided into 4 groups after interfering BANCR with lentiviral-mediated shRNA-1 and shRNA-2:interference group 1 (HCT116 cells transfected with LV-shRNA-1),interference group 2 (HCT116 cells transfected with LV-shRNA-2),negative control group (HCT116 cells transfected with lentivirus vector with nonsense sequence) and blank control group (HCT116 cells cultured in RPMI 1640 medium).The proliferation,apoptosis and migration of HCT116 cells in the 4 groups were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively.The comparison between the 2 groups was analyzed by u test,and multiple groups were compared by one-way analysis of variance,repeated measurement analysis of variance and LSD-t test.Multivariate analysis was done by Logistic regression model.The difference between categorical data was compared by chi-square test.Results The relative expression of BANCR in the cancer tissues was 1.6 ± 0.4,which was significantly higher than 0.9 ± 0.7 of the precancerous tissues (u =1 020.000,P ＜ 0.05).The result of univariate analysis showed that the high expression of BANCR was correlated with the lymph node metastasis and tumor stage (x2 =4.595,7.487,P ＜ 0.05).The result of multivariate analysis showed that lymph node metastasis and tumor stage (stage Ⅲ-Ⅳ) were the independent risk factors influencing the high expression of BANCR(OR =4.000,5.914,95％ CI:1.230-12.900,1.685-20.760,P ＜ 0.05).The relative expressions of BANCR of the interference group 1,interference group 2,negative control group and the blank control group were 0.25 ±0.04,0.20±0.06,0.96 ±0.04,0.98 ±0.03,with significant difference among the 4 groups (F =271.610,P ＜ 0.05).The cell proliferation rates at day 6 of the interference group 1,interference group 2 and the negative control group were 80.6％ ± 7.6％,81.2％ ± 5.1％ and 87.9％ ± 13.6％,with no significant difference among the 3 groups (F =0.559,P ＞ 0.05).The apoptotic rates of the interference group 1,interference group 2,negative control group and the blank control group were 4.7％ ± 1.7％,5.1％ ± 1.1％,3.1％ ± 0.6％ and 2.8％ ± 0.9％,with no significant difference among the 4 groups (F =2.881,P ＞ 0.05).The numbers of transmembrane cells of the interference group 1,interference group 2,negative control group and the blank control group were 135 ± 29,107 ± 18,240 ± 24 and 245 ± 22,with significant difference among the 4 groups (F =45.194,P ＜ 0.05).Conclusions BANCR was overexpressed in the HCT116 cells,and the BANCR overexpression was correlated with lymph node metastasis and tumor stage.BANCR can promote the migration of HCT116 cells.BANCR could be an important biomarker for the diagnosis and prognosis of colorectal cancer.

Molecular biology experiment is an important course for postgraduates in medical university.Through updating and optimizing what to teach(emphasizing study design,setting up comprehensive experiment and updating practical techniques),improving teaching mode(paying attention to experimental preparation,teacher qualification and postgraduates' capability training) and perfecting evaluation approaches(combining the experiment report with experimental summary and regular grades with experimental practice),good results have been obtained.

OBJECTIVETo explore the genetic causes for a child with multiple congenital malformations and epilepsy through analysis of copy number variations, and to correlate the genotype with the phenotype.

METHODSG-banding karyotyping was performed on the child and her parents. Single nucleotide polymorphisms array (SNP-array) was used to map the exact chromosomal breakpoints in the proband. The result was validated with fluorescence in situ hybridization (FISH).

RESULTSG banding analysis suggested that the proband had a karyotype of 46,XX,del(4)(p15), while both of his parents had a normal karyotype. SNP-array has identified a hemizygous deletion of 13.3 Mb on chromosome 4p16.3p15.33, which has been implicated in Wolf-Hirschhorn syndrome. FISH assay has confirmed the de novo origin of the deletion, with the karyotype and clinical phenotype of both parents taken into consideration.

CONCLUSIONA case of Wolf-Hirschhorn syndrome has been diagnosed by clinical manifestation and karyotyping analysis. Compared with conventional karyotyping analysis, SNP-array has greater resolution and accuracy, and can provide useful information for genetic counseling.

Chromosome Banding ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn ; Karyotyping ; Oligonucleotide Array Sequence Analysis ; Polymorphism, Single Nucleotide ; Wolf-Hirschhorn Syndrome ; genetics

Objective: Autologous lymph nodes fragmentary transplantation combined with vascular endothelial growth factor-C (VEGF-C) on athymic nude mice to explore the association between regeneration of lymphatic vessel and tumor cell migration. Methods: A total of 45 nude mice were randomly divided into 3 groups: Group A, simple autologous lymph nodes fragmentary transplantation, n=15; Group B, autologous lymph nodes fragmentary transplantation together with VEGF-C, n=15; Group C, without any intervention, n=15. At 1 month, 2 months and 6months after surgeries, the axillary lymph nodes of 5 mice in each group were dynamically examined by in vivo indocyanine green (ICG) fluorescence imaging respectively. The regenerated lymph nodes and relevant skin were evaluated using hematoxylin-eosin (H&E) staining and the skin was quantitatively analyzed via immunofluorescence staining for lymphatic vessel endothelial hyaluronan receptor 1(LYVE-1) as well. Results: One month after surgery, the right regenerated axillary lymph nodes in group B (5/5) were visible by in vivo ICG fluorescence imaging, whereas the same signals were not detected in group A (0/5). The results were the same at 2 and 6 months after surgery. HE staining showed that the cortical, paracortical, and medullary regions of the right axillary lymph nodes of the experimental group B were clear, and the lymphatic vessel structure was present, accompanied by lymphocyte infiltration. Immunofluorescence staining of the right upper limb showed that the expression of LYVE-1 in the lymphatic endothelium of the B group was significantly higher than that in group A (P<0.001) and the control group (both P<0.001). Conclusions Due to the promising consequence of regenerated lymph nodes, the procedure of autologous lymph nodes fragmentary transplantation combined with VEGF-C in athymic nude mice provides a reliable animal model for the next stage research.

Objective:To investigate the clinical effect of reconstructed tragus with hinge-shaped cartilage flap during the third stage reconstructive surgery in patients with microtia.Methods:From January 2018 to January 2019, the data of the patients with unilateral microtia of residual ear Type Ⅲ treated in the Seventh Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences were analyzed retrospectively. The residual ear tissue preserved in the second stage operation was used to reconstruct the tragus during the third stage operation, and the external auditory meatus was simulated with medium-thickness skin grafting. Postoperative changes were observed in the width, height, and symmetry of the tragus.Results:A total of 31 children with microtia were included, including 19 males and 12 females, aged from 6 to 14 years (mean 8 years), right ear ( n=16) and left ear ( n=15). Postoperative blood supply was good, and the incisions healed primarily. The patient’s immediate postoperative tragus width and height were slightly greater than the preoperative design requirements. After follow-up of 3-6 months, the width of the contralateral tragus was (1.05±0.12) cm, while the reconstructed tragus width was (0.98±0.17) cm. The difference was not statistically significant ( P=0.328). The height of the contralateral tragus was (0.53±0.08) cm, while the reconstructed tragus width was (0.55±0.08) cm. The difference was not statistically significant ( P=0.741). The reconstructed tragus had a good three-dimensional structure and stable shape, and the simulated shape of the external auditory meatus was good. Patients can wear earphones to meet the needs of daily life. Conclusions:Using residual ear cartilage to reconstruct the tragus by flapping hinge-cartilage can improve the three-dimensional structure of the auricle and skin grafting to the concha cavity posterior to the tragus can simulate external auditory meatus. The reconstructed ear resembles the healthy one and meets the need of wearing earphones.

Objective:To investigate the clinical effect of auricle reconstruction using an expanded postauricular flap with double pedicle in the ear reconstruction.Methods:From September 2016 to August 2017, the clinical data of all patients with congenital microtia treated in the Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences were analyzed retrospectively. The surgical procedures: The expander was implanted in the first stage. The ear framework was covered by the retroauricular double-pedicled expanded flap with free skin grafting at the retroauricular superior margin in the second stage, and the helix was wrapped by the retroauricular fascia. The patients were followed up at 1, 6 months after operation and before the third stage operation, and the appearance of the reconstructed auricle was evaluated by two junior doctors before the third stage of the operations. If there were any differences, the final evaluation would be carried out by a senior doctor. The evaluation results were good, medium and poor.Results:A total of 46 patients (49 ears) with congenital microtia were included, including 36 males (39 ears) and 10 females (10 ears), with the age range of 6-23 years. After operation, the blood supply was good, and the skin grafts survived completely. Three ears had postoperative hematomas under the double-pedicled expanded skin flap when the drainage was removed 5 days after operation. The hematomas recovered well after the negative pressure suction and dressing changed. During the follow-up of 6 months and 18 months, 43 cases (46 ears) were evaluated well with symmetrical and clear three-dimensional structure of reconstructed auricles, uniformed color, and hidden scar behind the ear. Three cases (3 ears) were evaluated as medium. There was hematoma when the drainage tube was removed 5 days after operation, which survived completely after negative suction and dressing change. No cartilage framework infection and exposure were found.Conclusions:The double-pedicled expanded flap has many advantages, such as reliable blood supply, natural auricle appearance, concealed scar, and less skin color difference. It is a good choice for soft tissue wrapping and covering during auricle reconstruction.

Objective:To construct a system HE-01 for the treatment of microtia based on HoloLens and verify the feasibility the application of HE-01 in ear reconstruction.Methods:Six volunteers and six patients with microtia were recruited from Plastic Surgery Hospital of Chinese Academy of Medical Sciences from August to September 2021. Mimics Research and 3-matic research software were used to construct a virtual three-dimensional auricle model. Based on HoloLens and occlusal splint navigation marker device, the virtual-real interactive system HE-01 for the treatment of microtia was constructed. The registration accuracy, tracking delay and display effect of HE-01 were evaluated through repeated experiments of volunteers. The operation process was defined through repeated experiments of patients and the feasibility of applying HE-01 in auricular reconstruction was verified.Results:HE-01 was built successfully and the operation flow was clear. The experiment of volunteers showed that under different angles, the registration accuracy was high (less than 2.7% of the bilateral auricle size difference of normal people), the tracking delay was low (less than 0.1s), and the green and red display effect of virtual auricle guide was the best. Experiments of patients with microtia showed that HE-01 could be applied to auricular reconstruction, and there were no obvious complications in wearing the navigation marker device, which had repeatability and stability.Conclusion:This study has completed the conceptual design of augmented-reality assisted auricular reconstruction based on navigation device, CT data and HoloLens. HE-01 has good registration accuracy, tracking speed and display effect, laying a good foundation for further software development and clinical transformation.

Objective:To construct a virtual-real interactive system HE-01 for the treatment of microtia based on HoloLens and verify the feasibility of applying HE-01 in ear reconstruction.Methods:Six volunteers (3 males and 3 females, average age: 20.5 years old) and six patients with microtia(3 males and 3 females, average age: 7.6 years old) were recruited from Plastic Surgery Hospital of Chinese Academy of Medical Sciences from August to September 2021. The Mimics Research 21.0 software and 3-matic research software were used to construct a virtual three-dimensional auricle model. Based on HoloLens and occlusal splint navigation marker device, the virtual-real interactive system HE-01 for the treatment of microtia was constructed. The registration accuracy, tracking delay and display effect of different colors (red, green, blue and skin color) of HE-01 were evaluated through repeated experiments of volunteers. The operation process was defined through repeated experiments of patients and the feasibility of applying HE-01 in auricular reconstruction was verified.Results:HE-01 was built successfully and the operation flow was clear. The experiment of six volunteers (12 ears) showed a high registration accuracy (average error rate 2.3%-2.4%, less than 2.7% of the bilateral auricle size difference of normal people), low tracking delay (six volunteers were all less than 0.1 s), and best green and red display effect of virtual auricle guide under different angles (-60°, -30°, 0°, 30°, 60°). Experiments of patients with microtia showed that HE-01 could be applied to auricular reconstruction, and there were no obvious complications in wearing the navigation marker device, which had repeatability and stability.Conclusion:This study has completed the conceptual design of augmented-reality assisted auricular reconstruction based on navigation device, CT data and HoloLens. HE-01 has good registration accuracy, tracking speed and display effect, laying a good foundation for further software development and clinical transformation.

Objective:To investigate the clinical effect of reconstructed tragus with hinge-shaped cartilage flap during the third stage reconstructive surgery in patients with microtia.Methods:From January 2018 to January 2019, the data of the patients with unilateral microtia of residual ear Type Ⅲ treated in the Seventh Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences were analyzed retrospectively. The residual ear tissue preserved in the second stage operation was used to reconstruct the tragus during the third stage operation, and the external auditory meatus was simulated with medium-thickness skin grafting. Postoperative changes were observed in the width, height, and symmetry of the tragus.Results:A total of 31 children with microtia were included, including 19 males and 12 females, aged from 6 to 14 years (mean 8 years), right ear ( n=16) and left ear ( n=15). Postoperative blood supply was good, and the incisions healed primarily. The patient’s immediate postoperative tragus width and height were slightly greater than the preoperative design requirements. After follow-up of 3-6 months, the width of the contralateral tragus was (1.05±0.12) cm, while the reconstructed tragus width was (0.98±0.17) cm. The difference was not statistically significant ( P=0.328). The height of the contralateral tragus was (0.53±0.08) cm, while the reconstructed tragus width was (0.55±0.08) cm. The difference was not statistically significant ( P=0.741). The reconstructed tragus had a good three-dimensional structure and stable shape, and the simulated shape of the external auditory meatus was good. Patients can wear earphones to meet the needs of daily life. Conclusions:Using residual ear cartilage to reconstruct the tragus by flapping hinge-cartilage can improve the three-dimensional structure of the auricle and skin grafting to the concha cavity posterior to the tragus can simulate external auditory meatus. The reconstructed ear resembles the healthy one and meets the need of wearing earphones.

Objective:To investigate the clinical effect of auricle reconstruction using an expanded postauricular flap with double pedicle in the ear reconstruction.Methods:From September 2016 to August 2017, the clinical data of all patients with congenital microtia treated in the Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences were analyzed retrospectively. The surgical procedures: The expander was implanted in the first stage. The ear framework was covered by the retroauricular double-pedicled expanded flap with free skin grafting at the retroauricular superior margin in the second stage, and the helix was wrapped by the retroauricular fascia. The patients were followed up at 1, 6 months after operation and before the third stage operation, and the appearance of the reconstructed auricle was evaluated by two junior doctors before the third stage of the operations. If there were any differences, the final evaluation would be carried out by a senior doctor. The evaluation results were good, medium and poor.Results:A total of 46 patients (49 ears) with congenital microtia were included, including 36 males (39 ears) and 10 females (10 ears), with the age range of 6-23 years. After operation, the blood supply was good, and the skin grafts survived completely. Three ears had postoperative hematomas under the double-pedicled expanded skin flap when the drainage was removed 5 days after operation. The hematomas recovered well after the negative pressure suction and dressing changed. During the follow-up of 6 months and 18 months, 43 cases (46 ears) were evaluated well with symmetrical and clear three-dimensional structure of reconstructed auricles, uniformed color, and hidden scar behind the ear. Three cases (3 ears) were evaluated as medium. There was hematoma when the drainage tube was removed 5 days after operation, which survived completely after negative suction and dressing change. No cartilage framework infection and exposure were found.Conclusions:The double-pedicled expanded flap has many advantages, such as reliable blood supply, natural auricle appearance, concealed scar, and less skin color difference. It is a good choice for soft tissue wrapping and covering during auricle reconstruction.