Objective To investigate the effect of nuclear factor erythroid-2-related factor 2(Nrf2) on the anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells(MSCs). Methods Human embryonic kidney cells(293FT) were transfected with recombinant plasmid which overexpressed Nrf2 and helper plasmid. High-titer lentivirus which overexpressed Nrf2 were obtained. MSCs were transfected with lentivirus with Nrf2 overexpression and empty lentiviral vector to establish Nrf2-MSCs which stably overexpressed Nrf2 (Nrf2 overexpression group) and green fluorescent protein (GFP)-MSCs(control group). The expression of green fluorescent in 2 groups was observed by fluorescence microscope. The expression level of Nrf2 protein in 2 groups was measured by Western Blot. The anti-hypoxia ability of 2 groups was observed by light microscope. The anti-apoptotic ability of 2 groups was measured by flow cytometry. Results Nrf2-MSCs which stably overexpressed Nrf2 were successfully established. Western Blot analysis revealed that the expression level of Nrf2 protein in the Nrf2 overexpression group was significantly higher than that in the control group(P<0.01). After 15 h hypoxia treatment, the cell activity in the Nrf2 overexpression group was significantly higher than that in the control group. Flow cytometry showed that the apoptosis rate in the Nrf2 overexpression group was (30.9±1.4)%, significantly lower than (61.3±1.3)% in the control group(P<0.05). Conclusions Nrf2-MSCs which can stably overexpress Nrf2 possess certain anti-hypoxia and anti-apoptotic ability in hypoxia environment.

Objective To establish a NOD/SCID mouse model with human immune reconstitution and observe its immune response to human triple-negative breast cancer xenograft. Methods Twenty-four NOD/SCID mice without immune leakage were subjected to cyclophosphamide (CTX) treatment 3 days prior to immune reconstitution with human peripheral blood mononuclear cell (PBMC) injection and subcutaneous transplantation of human triple-negative breast cancer MDA-MB-231 cells, CTX treatment and PBMC injection without tumor cell transplantation, MDA-MB-231 cell transplantation only, or no treatments. The tumor growth and immune responses of the mice were observed at regular intervals. Results Compared with the tumor-bearing mice, the tumor-bearing mice with immune reconstitution showed prolonged incubation period of tumor formation, slower tumor growth rate and increased survival rate. Human IgG and CD3+T cells were detected in the peripheral blood of the mice 1 week after human PBMC injection. The percentage of CD3+T cells in the spleen cells was 55.3%at 9 weeks in tumor-bearing mice with immune reconstitution and 52.7% in tumor-bearing mice without immune reconstitution. The spleen index of the tumor-bearing mice with immune reconstitution was much higher than that in mice with only immune reconstitution and the control mice (9.64 vs 3.82±0.31 and 1.51±0.14 mg/g). Conclusion A stable NOD/SCID mouse model with immune reconstitution has been established successfully, which shows immune responses to triple-negative breast cancer xenografts and allows studies of immunological therapy study of triple-negative breast cancer.

Objective To establish a NOD/SCID mouse model with human immune reconstitution and observe its immune response to human triple-negative breast cancer xenograft. Methods Twenty-four NOD/SCID mice without immune leakage were subjected to cyclophosphamide (CTX) treatment 3 days prior to immune reconstitution with human peripheral blood mononuclear cell (PBMC) injection and subcutaneous transplantation of human triple-negative breast cancer MDA-MB-231 cells, CTX treatment and PBMC injection without tumor cell transplantation, MDA-MB-231 cell transplantation only, or no treatments. The tumor growth and immune responses of the mice were observed at regular intervals. Results Compared with the tumor-bearing mice, the tumor-bearing mice with immune reconstitution showed prolonged incubation period of tumor formation, slower tumor growth rate and increased survival rate. Human IgG and CD3+T cells were detected in the peripheral blood of the mice 1 week after human PBMC injection. The percentage of CD3+T cells in the spleen cells was 55.3%at 9 weeks in tumor-bearing mice with immune reconstitution and 52.7% in tumor-bearing mice without immune reconstitution. The spleen index of the tumor-bearing mice with immune reconstitution was much higher than that in mice with only immune reconstitution and the control mice (9.64 vs 3.82±0.31 and 1.51±0.14 mg/g). Conclusion A stable NOD/SCID mouse model with immune reconstitution has been established successfully, which shows immune responses to triple-negative breast cancer xenografts and allows studies of immunological therapy study of triple-negative breast cancer.

BACKGROUND: Due to adipose-derived stem cells (ASCs) being easy to obtain, their rapid proliferation rate, and their multidirectional differentiation capabilities, they have been widely used in the field of regenerative medicine. With the progress of decellularized adipose tissue (DAT) and adipose tissue engineering research, the role of DAT in promoting angiogenesis has gradually been emphasized. METHODS: We examined the biological characteristics and biosafety of DAT and evaluated the stem cell maintenance ability and promotion of growth factor secretion through conducting in vitro and in vivo studies. RESULTS: The tested ASCs showed high rat:es of proliferation and adhered well to DAT. The expression levels of essential genes for cell stem maintenance, including OCT4, SOX2, and Nanog were low at 2–24 h and much higher at 48 and 96 h. The Adipogenic expression level of markers for ASCs proliferation including PPARγ, C/EPBα, and LPL increased from 2 to 96 h. Co-culture of ASCs and DAT increased the secretion of local growth factors, such as VEGF, PDGF-bb, bFGF, HGF, EGF, and FDGF-bb, and secretion gradually increased from 0 to 48 h. A model of full-thickness skin defects on the back of nude mice was established, and the co-culture of ASCs and DAT showed the best in vivo treatment effect. CONCLUSION The application of DAT promotes wound healing, and DAT combined with ASCs may be a promising material in adipose tissue engineering and regenerative medicine.

BACKGROUND: Due to adipose-derived stem cells (ASCs) being easy to obtain, their rapid proliferation rate, and their multidirectional differentiation capabilities, they have been widely used in the field of regenerative medicine. With the progress of decellularized adipose tissue (DAT) and adipose tissue engineering research, the role of DAT in promoting angiogenesis has gradually been emphasized. METHODS: We examined the biological characteristics and biosafety of DAT and evaluated the stem cell maintenance ability and promotion of growth factor secretion through conducting in vitro and in vivo studies. RESULTS: The tested ASCs showed high rat:es of proliferation and adhered well to DAT. The expression levels of essential genes for cell stem maintenance, including OCT4, SOX2, and Nanog were low at 2–24 h and much higher at 48 and 96 h. The Adipogenic expression level of markers for ASCs proliferation including PPARγ, C/EPBα, and LPL increased from 2 to 96 h. Co-culture of ASCs and DAT increased the secretion of local growth factors, such as VEGF, PDGF-bb, bFGF, HGF, EGF, and FDGF-bb, and secretion gradually increased from 0 to 48 h. A model of full-thickness skin defects on the back of nude mice was established, and the co-culture of ASCs and DAT showed the best in vivo treatment effect. CONCLUSION The application of DAT promotes wound healing, and DAT combined with ASCs may be a promising material in adipose tissue engineering and regenerative medicine.

Objective:To improve and optimize the modified surgical strategies for patents with Simon Ⅰ and Simon Ⅱ type of gynecomastia, based on their magnetic resonance imaging (MRI) features.Methods:Clinical data of 190 patients aged 12 to 56 years (mean age 26.7±8.6) with gynecomastia who underwent modified surgeries in the Department of Plastic Surgery of Peking Union Medical College Hospital from January 2017 to January 2023 were retrospectively analyzed. The study measured breast MRI images of 44 male patients to calculate the area ratio of the glands on the transverse plane via nipple. This provided insight into the physiological structure and the tissue distribution of gynecomastia for improving the modified surgical strategies. All 190 patients were treated with enhanced liposuction and " Pull-Through and Bottom-Up" techniques. The duration of operation, amount of liposuction and amount of glandular resection, postoperative complications were recorded, and postoperative satisfaction was investigated by questionnaire surveys.Results:MRI images of the 87-side male breast showed that the average area ratio of the glands on the transverse plane via nipple was (10.9±12.5) %. The most prevalent subtype of gynecomastia was branch pattern, accounting for 46.0%. The median surgical duration of 190 patients was 95 (65-210) minutes, the median liposuction volume was 300 (50-1 400) ml, and the median glandular removal was 19.9 (1.5-157.0) g. Eighty-eight patients (46.3%) went through followed up for more than 6 months post-posterative with an overall satisfaction score of 4.68±0.53. Postoperative complications occurred in 19 sides (5.1%), and the reoperation rate was 1.3%.Conclusions:The MRI imaging of gynecomastia shows that the component of patients′ breasts are mainly fat, with a small amount of glandular tissue and the dominant subtype is branch pattern. Enhanced liposuction combined with " Pull-Through and Bottom-Up" stab incision technique can be an effective treatment for Simon grades Ⅰ and Ⅱ gynecomastia. The method results in high patient satisfaction with fewer postoperative complications.

Objective:To evaluate the differences and aesthetic meaning of stable vitiligo treatment in the face and neck region using suction blister transplantation or noncultured autologous suspension of epidermal cells.Methods:Sixty-four stable vitiligo patients (25 males and 39 females with age ranges from 10 to 46 years, average 25 years) in the face and neck region were randomly divided into two groups (32 patients in each group): one group received suction blister transplantation, while other group received noncultured autologous suspension of epidermal cells. Patients′treatment effectiveness, pigmentation and piecing deformity were evaluated in postoperative 3 months and 6 months.Results:In the postoperative 3 months and 6 months, the effectiveness of suction blister transplantation group was 68.75% (22/32) and 90.63% (29/32), respectively, while the effectiveness of noncultured autologous suspension of epidermal cells group was 59.37% (19/32) and 87.50% (28/32), respectively, in which no significant differences were found between two groups ( P>0.05). No obvious pigmentation and piecing deformity were found in noncultured autologous suspension of epidermal cells group, which were much better than the suction blister transplantation group in postoperative 3 months and 6 months. Conclusions:Both suction blister transplantation and noncultured autologous suspension of epidermal cells could bring good treatment effectiveness for patients of stable vitiligo in the face and neck region. Compared with suction blister transplantation, noncultured autologous suspension of epidermal cells could offer better aesthetic appearance.

Objective:To investigate the clinical significance of suction blister transplantation in improving the efficacy of ReCell technique in the treatment of vitiligo.Methods:Patients were divided into three groups, namely, vitiligo patients without history of suction blister therapy, patients with ineffective suction blister therapy and patients with effective suction blister therapy. There were 30 patients in each group. All patients were treated with standard procedure of ReCell technique. The color recovery effect of leukoplakia was observed 3 and 6 months after operation, and the incidence of complications was also observed.Results:The effective rate of color recovery 3 and 6 months after operation were as follow: in patients without history of suction blister group, the effective rate of three months was 53.3%, and that of six months was 63.3%; in patients with ineffective suction blister group, the effective rate was 43.3% in three months and 50.0% in six months, and in patients with effective suction blister group, the effective rate was 76.7% in three months and 90.0% in six months. No obvious complications were observed in the three groups.Conclusions:For the treatment of stable vitiligo with ReCell technique, suction blister method is a simple and effective method for screening patients.

Objective To compare the effect of different size needle gauges to the degenerative response in rat caudal discs.Methods A total of 40 Sprague Dawley (SD) rats,level 5/6,7/8 and 9/10 interverbral discs of rat caudal spine were punctured with 18 or 21 or 25-gauge needles respectively.Radiographs and magnetic resonance imaging (MRI) were obtained at 1,2,4 and 6 weeks postsurgery.At each time point,ten rats from each group were sacrificed for histological analysis.Real-time fluorescent quantitative polymerase chain reaction (qPCR) was used to examine mRNA expression level.Results Significant differences were identified in the disc height index (DHI ％) and MRI grade between 18 G and normal group,MRI grade,histological score between 21 G and normal group at 2,4,and 6 weeks postsurgery.Significant differences were also identified in the histological score and mRNA expression levels between 18 G and normal group,alcian blue stain and hypoxia inducible factor-1α (HIF-1 α) mRNA expression level between 21G and normal group at all time point postsurgery.Significant differences existed in DHI％,type Ⅱ collagen and aggrecan mRNA expression levels between 21 G and normal group,all type mRNA expression levels between 25 G and normal group at 4,6 weeks.There were significant differences in MRI grade and histological score between 25 G and normal group at 6 weeks.Significant differences existed in almost all parameters compared between 18 G and 25 G at all time point.There were significant differences in DHI％,MRI grade,histological score and HIF-1α mRNA expression levels between 18 G and 21 G at 4,6 weeks.There were significant differences in type Ⅱ collagen and aggrecan mRNA expression levels between 18 G and 21 G at all time point.Significant differences exist in DHI％ and HIF-1α mRNA expression level between 21 G and 25 G at 6 weeks.Compared with the 25 G group,the DHI％ and Pfirrmann scores and the pathological score of each time at 2,4 and 6 weeks after operation in 18 G group have significant difference (P ＜ 0.05).Conclusions The middle size needle (21G) is better to induce disc degeneration.The 2-week time point may be the better time frame to further experimental treatments.

Objective:To compare the pros and cons of harvesting ear cartilage through anterior and posterior auricular approaches during rhinoplasty.Methods:From January 2017 to December 2018, 63 patients with otochondral rhinoplasty in our hospital were enrolled in this study, 60 were female and 3 were male; the average age was 31.6 years (range, 18 to 43) . They were randomly divided into anterior auricular approach group with 32 cases (64 sides) and posterior auricular approach group with 31 cases (62 sides). Surgical duration, complications and postoperative scar of the two methods were analyzed.Results:The average time for harvesting the ear cartilage through posterior auricular approach and anterior auricular approach was (20.8±1.7) min and (12.6±1.1) min, respectively ( P<0.01). The overall complication rate was 15.6% for posterior auricular approach group and 4.8% for anterior auricular approach group. The wound healed well in both groups, and there was no significant difference in postoperative scar between the two groups during an average 13 months follow-up period. Conclusions:While both the anterior and the posterior auricular approaches present with similar inconspicuous scarring, the use of anterior auricular approach alone to harvest ear cartilage during rhinoplasty provides both the surgeons and the patients with easier access, shorter surgical duration, and fewer complications. Hence, we believe that the anterior auricular approach possesses greater advantages than the posterior auricular approach.