Objective:To investigate the influence and mechanisms of metformin on the proliferation and apoptosis of human keloid fibroblasts (Fbs).Methods:This study was an experimental research. The keloid tissue was collected from 7 keloid patients (2 males and 5 females, aged 20-65 years, with a disease course of more than 1 year) who underwent keloid excision surgery at the Department of Plastic, Cosmetic and Maxillofacial Surgery of the First Affiliated Hospital of Xi'an Jiaotong University from September 2020 to September 2023. The primary Fbs were isolated and cultured, and cells from passages 3 to 6 were used for experiments. The cells were divided into control group and metformin group, and were cultured in complete medium. The medium for metformin group was supplemented with metformin at a final molarity of 60 mmol/L. The cell counting kit-8 was used to assess the proliferation activity of cells in two groups after 12 and 24 hours of culture, and the proliferation inhibition rate of cells in metformin group after 12 and 24 hours of culture was calculated, with a sample size of 6. The apoptosis detection kit was used to detect the apoptotic distribution of cells in control group after 0 hour (immediately) of culture and in metformin group after 12 and 24 hours of culture, with a sample size of 3. The cell cycle detection kit was used to detect the cycle distribution of cells in two groups after 12 and 24 hours of culture, with a sample size of 3. The eukaryotic mRNA sequencing was performed on suitable number of cells of two groups after 24 hours of culture, and the Kyoto encyclopedia of genes and genomes functional annotation analysis and functional enrichment analysis were performed after screening for differentially expressed genes (DEGs) with significantly differential expression between two groups. Western blotting was conducted to detect the protein expressions of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt), and phosphorylated mammalian target of rapamycin (p-mTOR) in the PI3K/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway of cells in two groups after 24 hours of culture, with a sample size of 3.Results:After 12 and 24 hours of culture, the proliferation activity of cells in metformin group was significantly lower than that in control group (with t values of 4.70 and 24.02, respectively, P<0.05); the proliferation activity of cells in metformin group after 24 hours of culture was significantly lower than that after 12 hours of culture within the group ( t=4.73, P<0.05). Compared with that after 12 hours of culture within the group, the proliferation inhibition rate of cells in metformin group was significantly increased after 24 hours of culture ( t=5.29, P<0.05). Compared with that in control group after 0 hour of culture, the proportion of early apoptotic cells in metformin group was significantly increased (with t values of 6.62 and 4.58, respectively, P<0.05), and the proportion of early and late apoptotic cells was significantly increased after 12 and 24 hours of culture (with t values of 4.84 and 3.75, respectively, P<0.05). After 24 hours of culture, the proportion of late apoptotic cells in metformin group was significantly higher than that after 12 hours of culture within the group ( t=4.55, P<0.05). After 12 hours of culture, the proportion of S-phase cells in metformin group was significantly lower than that in control group ( t=5.90, P<0.05). After 24 hours of culture, compared with that in control group, the proportion of G0/G1-phase cells in metformin group was significantly increased ( t=5.36, P<0.05), while the proportion of G2/M-phase cells was significantly decreased ( t=17.63, P<0.05). The proportion of S-phase cells in metformin group after 24 hours of culture was significantly higher than that after 12 hours of culture within the group ( t=7.60, P<0.05). After 24 hours of culture, 4 814 DEGs with significantly differential expression were detected in the cells of metformin group compared with control group. The significantly upregulated and downregulated DEGs were mainly involved in biological functions related to signal transduction, cell growth and death, transport and catabolism, the endocrine system, the immune system, and cancer. The pathways that were significantly enriched with DEGs with significantly differential expression included the cell cycle and DNA replication, with the highest number of genes in the PI3K/Akt signaling pathway. After 24 hours of culture, the protein expressions of PI3K, p-Akt, and p-mTOR of cells in metformin group were 0.190±0.017, 0.170±0.017, and 0.247±0.005, respectively, which were significantly lower than 0.440±0.026, 0.300±0.060, and 0.547±0.025 in control group (with t values of 13.69, 3.61, and 20.12, respectively, P values all <0.05). Conclusions:Metformin can significantly inhibit the proliferation of human keloid Fbs through the PI3K/Akt/mTOR signaling pathway and effectively induce its apoptotic process, thereby exerting antifibrotic effects.

Objective:To investigate the influence and mechanisms of metformin on the proliferation and apoptosis of human keloid fibroblasts (Fbs).Methods:This study was an experimental research. The keloid tissue was collected from 7 keloid patients (2 males and 5 females, aged 20-65 years, with a disease course of more than 1 year) who underwent keloid excision surgery at the Department of Plastic, Cosmetic and Maxillofacial Surgery of the First Affiliated Hospital of Xi'an Jiaotong University from September 2020 to September 2023. The primary Fbs were isolated and cultured, and cells from passages 3 to 6 were used for experiments. The cells were divided into control group and metformin group, and were cultured in complete medium. The medium for metformin group was supplemented with metformin at a final molarity of 60 mmol/L. The cell counting kit-8 was used to assess the proliferation activity of cells in two groups after 12 and 24 hours of culture, and the proliferation inhibition rate of cells in metformin group after 12 and 24 hours of culture was calculated, with a sample size of 6. The apoptosis detection kit was used to detect the apoptotic distribution of cells in control group after 0 hour (immediately) of culture and in metformin group after 12 and 24 hours of culture, with a sample size of 3. The cell cycle detection kit was used to detect the cycle distribution of cells in two groups after 12 and 24 hours of culture, with a sample size of 3. The eukaryotic mRNA sequencing was performed on suitable number of cells of two groups after 24 hours of culture, and the Kyoto encyclopedia of genes and genomes functional annotation analysis and functional enrichment analysis were performed after screening for differentially expressed genes (DEGs) with significantly differential expression between two groups. Western blotting was conducted to detect the protein expressions of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt), and phosphorylated mammalian target of rapamycin (p-mTOR) in the PI3K/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway of cells in two groups after 24 hours of culture, with a sample size of 3.Results:After 12 and 24 hours of culture, the proliferation activity of cells in metformin group was significantly lower than that in control group (with t values of 4.70 and 24.02, respectively, P<0.05); the proliferation activity of cells in metformin group after 24 hours of culture was significantly lower than that after 12 hours of culture within the group ( t=4.73, P<0.05). Compared with that after 12 hours of culture within the group, the proliferation inhibition rate of cells in metformin group was significantly increased after 24 hours of culture ( t=5.29, P<0.05). Compared with that in control group after 0 hour of culture, the proportion of early apoptotic cells in metformin group was significantly increased (with t values of 6.62 and 4.58, respectively, P<0.05), and the proportion of early and late apoptotic cells was significantly increased after 12 and 24 hours of culture (with t values of 4.84 and 3.75, respectively, P<0.05). After 24 hours of culture, the proportion of late apoptotic cells in metformin group was significantly higher than that after 12 hours of culture within the group ( t=4.55, P<0.05). After 12 hours of culture, the proportion of S-phase cells in metformin group was significantly lower than that in control group ( t=5.90, P<0.05). After 24 hours of culture, compared with that in control group, the proportion of G0/G1-phase cells in metformin group was significantly increased ( t=5.36, P<0.05), while the proportion of G2/M-phase cells was significantly decreased ( t=17.63, P<0.05). The proportion of S-phase cells in metformin group after 24 hours of culture was significantly higher than that after 12 hours of culture within the group ( t=7.60, P<0.05). After 24 hours of culture, 4 814 DEGs with significantly differential expression were detected in the cells of metformin group compared with control group. The significantly upregulated and downregulated DEGs were mainly involved in biological functions related to signal transduction, cell growth and death, transport and catabolism, the endocrine system, the immune system, and cancer. The pathways that were significantly enriched with DEGs with significantly differential expression included the cell cycle and DNA replication, with the highest number of genes in the PI3K/Akt signaling pathway. After 24 hours of culture, the protein expressions of PI3K, p-Akt, and p-mTOR of cells in metformin group were 0.190±0.017, 0.170±0.017, and 0.247±0.005, respectively, which were significantly lower than 0.440±0.026, 0.300±0.060, and 0.547±0.025 in control group (with t values of 13.69, 3.61, and 20.12, respectively, P values all <0.05). Conclusions:Metformin can significantly inhibit the proliferation of human keloid Fbs through the PI3K/Akt/mTOR signaling pathway and effectively induce its apoptotic process, thereby exerting antifibrotic effects.

Objective:To investigate the clinical effect of repairing unilateral incomplete cleft lip functionally and sub-regionally with modified micro-triangular skin flap and orbicularis oris muscle flaps.Methods:A retrospective study was conducted in the patients with unilateral incomplete cleft lip, who underwent repair at Department of Plastic, Aesthetic and Maxillofacial Surgery, the First Affiliated Hospital of Xi’an Jiaotong University from April 2020 to February 2023. Surgical landmarks were fixed according to the anatomical structure of non-cleft side. Short straight skin incisions were designed along the philtral column. A micro-triangular flap was designed above the vermilion margin of the affected lip, which was inserted into the contralateral lip peak to lower the lip peak and lengthen the philtral column. The orbicularis oris muscle was reconstructed with five muscle flaps in three areas to create a good sub-structure of the upper lip and the nasal floor. The vermilion tubercle and philtral column were reconstructed. Deviation of nasal columella was corrected and the nasal floor was elevated. The outcomes were assessed through subjective evaluation and objective anthropometric measurements.(1) Subjective outcomes were assessed by two plastic surgeons together who were not included in this study. The following parameters were assessed: scar appearance, Cupid’s bow continuity, lip pick height, alar base width, nostril symmetry, philtral ridge contour. Each parameter was graded from 1 point (poor), 2 points (average), or 3 points (good). (2) Objective measurements were taken by one plastic surgeon who was not included in this study using the Image-Pro Plus 6.0. Measurements were included bilateral vermilion thickness, bilateral length of lip pick to cheilion, bilateral philtral column length, bilateral length of Cupid’s pick to ala nasi, bilateral alar base width. Asymmetry ratio = |non-cleft counts-cleft counts|/non-cleft counts×100%, and a value closer to 0 would mean the less different, the more symmetrical. Statistical analysis was performed using descriptive methods. Non-normal distributed measurement datas were expressed by M( Q1, Q3). Results:A total of 32 patients of unilateral incomplete cleft lip were enrolled, including 19 males and 13 females, aged 3-18 months. All patients were primary healing, and no serious complications (i.e., infection, hematoma, wound dehiscence) occurred. The postoperative follow-up time was 6-24 months. The patients were satisfied with the results, including favorable red lip contour, good continuity, obvious vermilion, cubical philtrum column, good symmetry of bilateral structure and sub-structure and light scar. The overall score of the subjective evaluation was 2.66 points. Cupid’s bow continuity got the highest score(2.84 points), and nostril symmetry got the lowest score(2.38 points). Objective measurements indicated excellent parameters were bilateral alar base width [2.60%(1.02%, 7.08%)] and bilateral philtral length[3.95%(2.03%, 5.98%)].Conclusion:Repairing unilateral incomplete cleft lip functionally and sub-regionally with modified micro-triangular skin flap and orbicularis oris muscle flaps can create a good sub-structure of the upper lip contour, and bring a significant improvement in the upper lip and the nasal floor symmetry, which is an effective method for incomplete unilateral cleft lip repair.

Objective:To investigate the clinical effect of repairing unilateral incomplete cleft lip functionally and sub-regionally with modified micro-triangular skin flap and orbicularis oris muscle flaps.Methods:A retrospective study was conducted in the patients with unilateral incomplete cleft lip, who underwent repair at Department of Plastic, Aesthetic and Maxillofacial Surgery, the First Affiliated Hospital of Xi’an Jiaotong University from April 2020 to February 2023. Surgical landmarks were fixed according to the anatomical structure of non-cleft side. Short straight skin incisions were designed along the philtral column. A micro-triangular flap was designed above the vermilion margin of the affected lip, which was inserted into the contralateral lip peak to lower the lip peak and lengthen the philtral column. The orbicularis oris muscle was reconstructed with five muscle flaps in three areas to create a good sub-structure of the upper lip and the nasal floor. The vermilion tubercle and philtral column were reconstructed. Deviation of nasal columella was corrected and the nasal floor was elevated. The outcomes were assessed through subjective evaluation and objective anthropometric measurements.(1) Subjective outcomes were assessed by two plastic surgeons together who were not included in this study. The following parameters were assessed: scar appearance, Cupid’s bow continuity, lip pick height, alar base width, nostril symmetry, philtral ridge contour. Each parameter was graded from 1 point (poor), 2 points (average), or 3 points (good). (2) Objective measurements were taken by one plastic surgeon who was not included in this study using the Image-Pro Plus 6.0. Measurements were included bilateral vermilion thickness, bilateral length of lip pick to cheilion, bilateral philtral column length, bilateral length of Cupid’s pick to ala nasi, bilateral alar base width. Asymmetry ratio = |non-cleft counts-cleft counts|/non-cleft counts×100%, and a value closer to 0 would mean the less different, the more symmetrical. Statistical analysis was performed using descriptive methods. Non-normal distributed measurement datas were expressed by M( Q1, Q3). Results:A total of 32 patients of unilateral incomplete cleft lip were enrolled, including 19 males and 13 females, aged 3-18 months. All patients were primary healing, and no serious complications (i.e., infection, hematoma, wound dehiscence) occurred. The postoperative follow-up time was 6-24 months. The patients were satisfied with the results, including favorable red lip contour, good continuity, obvious vermilion, cubical philtrum column, good symmetry of bilateral structure and sub-structure and light scar. The overall score of the subjective evaluation was 2.66 points. Cupid’s bow continuity got the highest score(2.84 points), and nostril symmetry got the lowest score(2.38 points). Objective measurements indicated excellent parameters were bilateral alar base width [2.60%(1.02%, 7.08%)] and bilateral philtral length[3.95%(2.03%, 5.98%)].Conclusion:Repairing unilateral incomplete cleft lip functionally and sub-regionally with modified micro-triangular skin flap and orbicularis oris muscle flaps can create a good sub-structure of the upper lip contour, and bring a significant improvement in the upper lip and the nasal floor symmetry, which is an effective method for incomplete unilateral cleft lip repair.

Objective To study the effect of autogeneic platelet-rich fibrin (PRF) on proliferation and adipogenic differentiation of human adipose-derived stem cells (ADSCs) in vitro.Methods ADSCs were isolated from adipose tissue obtained from donors undergoing liposuction and were cultured,and underwent identification.ADSCs at passage 3 were divided into three groups:test groups were cultured with 1PRFM and 2PRFM,and control group was cultured without PRF membrane.Then the growth of the cells was observed by inverted microscope.MTT method was used to observe cell proliferation activity at days 1,2,3,4,5,6 and 7 after culture.Adipogenic differentiation of ADSCs was observed and quantified by oil red O staining at days 3,5,7,9,11 and 14.Results Cell proliferation and adipogenic differentiation would be increased with the PRFM,There were significant differences among three groups.Conclusions PRF could significantly promote proliferation and adipogenic differentiation of ADSCs.

Objective To explore a better method to repair cervical cicatricle contracture deformity.Methods The cervicai cicatrical contraction deformity was repaird by the cervico-shoulder island skin flap with blood supply crossing and abundant anastomotic branches of the neck-shoulder blood vessels. After removing the cervical scar, the flaps were designed according to the size of the wound to be repaired, and the clinical anatomy of crossing and abundant anastomotic branches of the neck-shoulder blood vessels. The secondary wound of donor site was closed directly if the donor region was small, or closed by skin graft. Results Sixteen patients in this clinic were operated with this method, and venous return dysfunction occurred in 1 case, but blood circulation was improved after treatment. All the flaps survived with good appearance and texture. The extensive flap was created and transferred to the anterior neck by one stage without pre-expansion which met some patients'requirement of repairing cervical cicatricle contracture deformity. Conclusion The cervico-shoulder island skin flap, with abundant blood supply, is a new method to repair cervical cicatricle contracture deformity.

Objective To explore the effect of body-weight support treadmill training (BWSTT)on ambulatory capacity in individuals following incomplete spinal cord injury (ISCI).Methods 36 cases of incomplete SCI patients were divided randomly into control and BWSTT groups who were classified as ASIA C or D.All received comprehensive rehabilitation, and patients in BWSTT group also received BWSTT for 3 months when the physical condition was permitted.They were assessed before and after treatment.The evaluation items included measurement of ASIA lower-extremity motor assessment, functional independence measure (FIM) ambulatory function measure, 10meter ambulatory velocity and modified Barthel Index.Results The scores of ASIA lower-extremity motor assessment, FIM ambulatory function, the ambulatory 10-meter velocity in BWSTT group improved significantly when compared with the control group (P<0.05).The scores of modified Barthel Index were no difference between two groups before and after treatment (P<0.05).Conclusion BWSTT could improve ambulatory capacity for incomplete SCI patients

Objective To observe the effects of comprehensive rehabilitation treatment on ambulation and balance function in patients with heavy brain injury when the starting time of rehabilitation was delayed.Methods Twelve heavy brain-injured patients received comprehensive rehabilitation treatment,including ambulation and balance training.They were evaluated with Fugl-Meyer Lower Extremity Assessment Scale,Functional Independence Measure(FIM)Ambulation Function Scale,Berg Balance Scale and Modified Barthel Index(MBI).One-way ANOVA was used to assess the functional status of patients during the course of treatment.Results After 3～6 months of treatment,All the scores of the assessment items above scales in all of the patients increased significantly compared with those at first evaluation(P<0.05).Conclusion The heavy brain-injured patients can still acquire contented ambulation and balance function after accepting comprehensive rehabilitation,though the starting time of rehabilitation is delayed.