Objective:To classify the extensive defects after upper lip cancer surgery and explore the efficacy of nasolabial flap with Abbe flap combined or vermilion elastic flap in extensive defects after upper lip cancer ablation.Methods:Twenty-seven patients with upper lip cancer who underwent surgical resection in the Department of Oral and Maxillofacial, The Second Xiangya Hospital, Central South University from January 2010 to December 2021 were recruited, including 13 males and 14 females aged from 19 to 78 years old. The defects of upper lip were more than 50%, and local flaps (Abbe flap, nasolabial flap or vermilion elastic flap) were used to reconstruct the defects of upper lip cancer. According to the extent of defects and the application type of flaps, the defects were classified into 3 sub-types: ①Type Ⅰ defect: median upper lip defect, with more than 50% defect of vermilion and nearly complete defect of upper lip skin, which can be repaired by Abbe flap combined with double nasolabial flap.②Type Ⅱ defect: median upper lip defect, with more than 50% defect of vermilion and upper lip skin defect, repaired by Abbe flap alone. ③Type Ⅲ defect: lateral upper lip defect, with more than 50% defect of vermilion and upper lip skin defect. The nasolabial flap was used to reconstruct the skin defect, and the vermilion elastic flap was used to repair the labial redness defect. A four-point scale was used to evaluate the function and aesthetic outcome of the lips 3 months after surgery, and postoperative complications were recorded.Results:Among the included patients with lip cancer, there were 2 cases of type Ⅰ, 9 cases of type Ⅱ, and 16 cases of type Ⅲ. All Abbe flap and nasolabial flap survived, with tip necrosis occurring in 2 Abbe flaps and have been resected. The average functional score was 2.89±0.80, and the average cosmetic score was 3.04±0.76.Conclusions:The reconstructive method based on Abbe flap and nasolabial flap provides a novel surgical technique for the reconstruction of extensive defects after upper lip cancer surgery.

Objective:To analyze the prognostic factors of tongue squamous cell carcinoma(TSCC)and provide a clinical refer-ence for the treatment of TSCC.Methods:329 patients underwent radical resection of TSCC primary tumor and neck dissection were included in this study.Results:The overall 5-year survival rate was 73.01％and the local control rate was 84.74％.Cox multivari-ate analysis showed that age(P=0.045 1),histological grade(P＜0.000 1)and clinical stage(P=0.004 9)were closely related to the 5-year overall survival rate of the patients with TSCC.Multivariate linear regression analysis showed that histopathological grade(P=0.000 3),T stage(P=0.009 7)and lymph node metastasis(P=0.040 6)was closely related to local disease control in the patients.Conclusion:Age,histological grade and clinical stage are important prognostic factors,the histological grade,T stage and lymph node metastasis are closely related to local disease control in patients with TSCC.

Objective:To classify the extensive defects after upper lip cancer surgery and explore the efficacy of nasolabial flap with Abbe flap combined or vermilion elastic flap in extensive defects after upper lip cancer ablation.Methods:Twenty-seven patients with upper lip cancer who underwent surgical resection in the Department of Oral and Maxillofacial, The Second Xiangya Hospital, Central South University from January 2010 to December 2021 were recruited, including 13 males and 14 females aged from 19 to 78 years old. The defects of upper lip were more than 50%, and local flaps (Abbe flap, nasolabial flap or vermilion elastic flap) were used to reconstruct the defects of upper lip cancer. According to the extent of defects and the application type of flaps, the defects were classified into 3 sub-types: ①Type Ⅰ defect: median upper lip defect, with more than 50% defect of vermilion and nearly complete defect of upper lip skin, which can be repaired by Abbe flap combined with double nasolabial flap.②Type Ⅱ defect: median upper lip defect, with more than 50% defect of vermilion and upper lip skin defect, repaired by Abbe flap alone. ③Type Ⅲ defect: lateral upper lip defect, with more than 50% defect of vermilion and upper lip skin defect. The nasolabial flap was used to reconstruct the skin defect, and the vermilion elastic flap was used to repair the labial redness defect. A four-point scale was used to evaluate the function and aesthetic outcome of the lips 3 months after surgery, and postoperative complications were recorded.Results:Among the included patients with lip cancer, there were 2 cases of type Ⅰ, 9 cases of type Ⅱ, and 16 cases of type Ⅲ. All Abbe flap and nasolabial flap survived, with tip necrosis occurring in 2 Abbe flaps and have been resected. The average functional score was 2.89±0.80, and the average cosmetic score was 3.04±0.76.Conclusions:The reconstructive method based on Abbe flap and nasolabial flap provides a novel surgical technique for the reconstruction of extensive defects after upper lip cancer surgery.

Objective:To analyze the prognostic factors of tongue squamous cell carcinoma(TSCC)and provide a clinical refer-ence for the treatment of TSCC.Methods:329 patients underwent radical resection of TSCC primary tumor and neck dissection were included in this study.Results:The overall 5-year survival rate was 73.01％and the local control rate was 84.74％.Cox multivari-ate analysis showed that age(P=0.045 1),histological grade(P＜0.000 1)and clinical stage(P=0.004 9)were closely related to the 5-year overall survival rate of the patients with TSCC.Multivariate linear regression analysis showed that histopathological grade(P=0.000 3),T stage(P=0.009 7)and lymph node metastasis(P=0.040 6)was closely related to local disease control in the patients.Conclusion:Age,histological grade and clinical stage are important prognostic factors,the histological grade,T stage and lymph node metastasis are closely related to local disease control in patients with TSCC.

Objective:To explore the mechanism of miR-15b-5p involving in depression-like behavior in Parkinson's disease (PD).Methods:(1) Eighteen C57BL/6N mice were randomly divided into PD group, intervention group and control group ( n=6). PD models in PD group were established by stereotaxically injecting 0.25 mg/kg rotenone into the right striatum; mice in the intervention group were injected with 0.25 mg/kg rotenone and miR-15b-5p inhibitor lentivirus, while mice in the control group were injected with equal volume of PBS. Four weeks after that, open field test and rotarod test were performed to evaluate the motor ability, and sucrose preference and forced swimming tests were performed to evaluate the depression-like behaviors. And then, proteins and miRNAs in the substantia nigra were extracted; real-time fluorescent quantitative reverse transcription PCR (qRT-PCR) was used to detect the miR-15b-5p expression,and Western blotting and immunofluorescent staining were used to detect the tyrosine hydroxylase (TH), brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), and postsynaptic density protein 95 (PSD95) protein expressions. (2) The 100 nmol/L miR-15b-5p mimic/inhibitor and their negative control sequences were transfected into SH-SY5Y cells on 6-well plates (named miR-15b-5p mimic group, miR-15b-5p mimic control group, miR-15b-5p inhibitor group and miR-15b-5p inhibitor control group, respectively); 48 h after that, BDNF, TrkB and PSD95 protein expressions were detected by Western blotting and miR-15b-5p expression by qRT-PCR. (3) The 100 ng BDNF 3'-UTR wild-type or mutant luciferase reporter vector plasmids and 100 nmol/L miR-15b-5p mimic/inhibitor or their negative control sequences were co-transfected into SH-SY5Y cells on 24-well plates, and luciferase reporter activity assay was performed 48 h after co-transfection to detect the luciferase activity. Results:(1) Compared with the control group, the PD group had significantly reduced movement speed, shortened rotarod drop latency, decreased percentage of sucrose preference, and prolonged immobility time ( P<0.05); compared with the PD group, the intervention group had significantly increased movement speed, prolonged rotarod drop latency, increased percentage of sucrose preference, and shortened immobility time ( P<0.05). (2) Compared with the miR-15b-5p mimic control group, the miR-15b-5p mimic group had significantly increased miR-15b-5p expression, and decreased BDNF, TrkB and PSD95 protein expressions (100.00±5.75 vs. 66.79±5.90; 100.00±5.95 vs. 84.46±5.77; 100.00±7.02 vs. 80.43±3.25, P<0.05). Compared with the miR-15b-5p inhibitor control group, the miR-15b-5p inhibitor group had significantly decreased miR-15b-5p expression, and increased BDNF, TrkB and PSD95 expressions (100.00±6.81 vs. 119.90±5.66; 100.00±2.88 vs. 110.10±4.15; 100.00±2.19 vs. 124.60±11.69, P<0.05). Compared with the control group, PD group had significantly increased miR-15b-5p expression, and significantly decreased TH, BDNF, TrkB and PSD95 expressions and BDNF fluorescent intensity (100.00±9.20 vs. 63.60±12.80; 100.00±9.88 vs. 71.95±10.00; 100.00±5.16 vs. 70.37±8.43; 100.00±7.01 vs. 68.12±10.22; 100.00±12.99 vs. 48.23±12.58) in the substantia nigra ( P<0.05); compared with the PD group, the intervention group had significantly lower miR-15b-5p expression and increased TH, BDNF, TrkB and PSD95 expressions and BDNF fluorescent intensity (63.60±12.80 vs. 90.69±9.84; 71.95±10.00 vs. 93.31±4.50; 70.37±8.43 vs. 88.11±4.10; 68.12±10.22 vs. 89.59±5.93; 48.23±12.58 vs. 83.65±10.52) in the substantia nigra ( P<0.05). (3) Compared with the BDNF 3'-UTR wild-type+miR-15b-5p mimic control group, the BDNF 3'-UTR wild-type+miR-15b-5p mimic group had significantly decreased luciferase activity (100.00±5.07 vs. 90.59±1.75, P<0.05); compared with the BDNF 3'-UTR wild-type+miR-15b-5p inhibitor control group, the BDNF 3'-UTR wild-type+miR-15b-5p inhibitor group had significantly increased luciferase activity (100.00±5.08 vs. 152.20±31.87, P<0.05). Conclusion:MiR-15b-5p is involved in depression-like behavior in PD by down-regulating the BDNF/TrkB/PSD95 expressions.

Objective:To observe the effect of α-synuclein (α-Syn) aggregates incubated with plasma from patients with Parkinson's disease (PD) and multiple system atrophy (MSA) on mitochondrial and lysosomal functions, and further clarify the role of α-Syn in pathological mechanism of PD and MSA.Methods:Human neuroblastoma cells (SH-SY5Y) were treated with 10 μg/mL α-Syn aggregates incubated with phosphate buffered solution (PBS), and plasma from healthy controls (HCs), PD patients, and MSA patients, respectively; and the untreated SH-SY5Y cells were used as control group. Mito-Tracker Red CMXRos fluorescent staining was used to detect the mitochondrial membrane potential, Western blotting was used to detect the cytochrome C (Cyt C) expression in the mitochondria and cytoplasm, and adenosine triphosphate (ATP) test kit was used to detect the intracellular ATP level. In addition, the lysosomal associated membrane protein-1 (LAMP1) expression was detected by Western blotting, and cathepsin B activity was detected by Magic Red fluorescent staining.Results:(1) Effects of different α-Syn aggregates on mitochondrial function damage: compared with the control group, the PBS, HC, PD and MSA groups had significantly decreased relative fluorescent intensity (Mito-Tracker Red CMXRos staining) in the SH-SY5Y intracellular mitochondria ( P<0.05); the PD group and MSA group had significantly decreased relative fluorescent intensity compared with PBS group and HC group ( P<0.05); the MSA group had significantly decreased relative fluorescent intensity compared with the PD group (41.27%±5.97% vs. 60.05%±4.24%, P<0.05). Compared with the control group, the HC, PD and MSA groups had significantly decreased mitochondrial Cyt C protein expression, while the PBS, HC, PD and MSA groups had significantly increased cytoplasmic Cyt C protein expression ( P<0.05); the PD group and MSA group had significantly decreased mitochondrial Cyt C protein expression compared with the PBS group and HC group ( P<0.05); the MSA group had significantly increased cytoplasmic Cyt C protein expression compared with the PBS group ( P<0.05); the MSA group had significantly decreased mitochondrial Cyt C protein expression (51.50%±4.22%) and increased cytoplasmic Cyt C protein expression (199.40%±6.73%) compared with the PD group (65.52%±2.18% and 174.00%±16.18%, P<0.05). Compared with the control group, the PBS group, HC group, PD group and MSA group had significantly decreased ATP level ( P<0.05); the PD group and MSA group had significantly decreased ATP level compared with the PBS group and HC group ( P<0.05); the MSA group had significantly decreased ATP level (1.81±0.13 nmol/mg) compared with PD group (2.96±0.29 nmol/mg, P<0.05). (2) Effects of different α-Syn aggregates on lysosomal functions: compared with the control group, the PD group and MSA group had significantly decreased LAMP1 protein expression ( P<0.05); the MSA group had significantly decreased LAMP1 protein expression (60.74%±7.94%) compared with the PBS group, HC group, and PD group (79.70%±4.34%, P<0.05). Compared with the control group, the PBS, HC, PD and MSA groups had significantly enhanced relative fluorescent intensity of lysosomal cathepsin B ( P<0.05); the PD group and MSA group had significantly enhanced relative fluorescent intensity of lysosomal cathepsin B compared with PBS group and HC group ( P<0.05); the MSA group had significantly increased relative fluorescent intensity of lysosomal cathepsin B compared with PD group (399.00%±35.54% vs. 305.90%±24.87%, P<0.05). Conclusion:The α-Syn aggregates incubated with plasma from PD and MSA patients have damage to mitochondrial and lysosome functions, especially those incubated with plasma from MSA patients.

Objective:To analyze the characteristics of cervical lymph node metastasis in tongue squamous cell carcinoma (TSCC).Methods:A retrospective study was conducted. A total of 329 patients with TSCC who underwent en bloc resection of primary tumor and neck dissection in the Second Xiangya Hospital of Central South University from June 2010 to March 2018 were included. There were 283 males and 46 females, aged from 26 to 80 years. All patients underwent the modified neck dissection. The main difference between the modified neck dissection and the traditional neck dissection lay in the managements of unconventional lymph nodes. The lymphatic adipose tissues adjacent to the superior thyroid artery, the base of facial artery and the branches of external carotid artery were thoroughly dissected. The primary tumor as well as lingual artery, tissues along the lingual artery and lymph nodes in the mouth floor were resected. χ 2 test was used for comparison of count data, and linear regression model was used for multivariate analysis. Results:Cervical lymph node metastases were found in 136 patients (41.3%). Among 142 patients (T1-2cN0) with supraomohyoid neck dissection, 22 patients had pathologically occult lymph node metastases (15.5%), with a 5-year overall survival rate of 90.2%, which was similar to the 5-year overall survival rate of 92.1% in 120 patients without lymph node metastasis (χ 2=0.156, P=0.693). Multivariate linear regression analysis showed that T stage, clinical stage and unconventional lymph node metastasis were important factors for cervical lymph node metastasis in tongue cancer patients ( P<0.05). Unconventional lymph node metastases occurred in 30 patients (9.1%), including the metastases of lymph nodes in the floor of mouth (3.0%), the lingual artery (2.4%), the base of the external maxillary artery (2.1%), the superior thyroid artery (0.9%), and the external carotid artery (0.6%). There were significant differences in the unconventional lymph node metastasis rates between patients with negative and positive conventional lymph node metastases [4.9%(10/203) vs. 15.9%(20/126), χ 2=11.242, P=0.001] and also between patients with depth of invasion ≤5 mm, 5 mm 10 mm [3.1%(2/64) vs. 5.7%(6/106) vs. 13.8%(22/159), χ 2=7.907, P=0.005]. Conclusion:Supraomohyoid neck dissection can achieve reliable control efficacy in patients with cN0 tongue cancer. All patients with lymph node dissection should undergo unconventional lymph node dissection. Unconventional lymph node dissection is strongly recommended for patients with conventional lymph node metastasis.

Objective:To investigate the effect of anatomical unit resection on the overall survival rate of patients with T4 stage posterior buccal carcinoma.Methods:We conducted a retrospective study on 79 patients with T4 stage posterior buccal squamous cell carcinoma underwent radical surgery for buccal mucosa cancer and reconstruction with free anterolateral thigh flap in the Department of Oral and Maxillofacial Surgery, the Second Xiangya Hospital, Central South University from March 2014 to December 2019. The 74 patients were males and the 5 patients were females, aged from 28 to 72 years old. The 40 patients in control group underwent traditional extended resection (1.5-2.0 cm safe margin), and 39 patients in the experimental group underwent anatomical unit resection. The clinicopathological parameters of patients were collected and followed up regularly. The overall survival rate was assessed by Log-rank and Cox proportional risk regression model. Multivariate analysis was performed by linear regression model.Results:There were significant differences between experimental group and control group in overall survival rates (61.53% vs 27.50%, χ2=6.624, P=0.010) and local disease control rates (74.36% vs 27.50%, χ2=17.350, P<0.001). Multiple linear regression analysis showed that local disease control rate was significantly correlated with anatomical unit resection ( t=3.880, P<0.001) and lymph node metastasis ( t=2.619, P=0.011), and also Cox proportional hazards regression model analysis identified that overall survival rate was significantly correlated with anatomical unit resection ( Z=2.421, P=0.016) and lymph node metastasis ( Z=2.793, P=0.005). Conclusion:For posterior buccal carcinoma with multiple anatomical units involved, anatomical unit resection can significantly reduce local recurrence and improve overall survival rate of patients.

Objective:To observe the effect of α-synuclein (α-Syn) aggregates incubated with plasma from patients with Parkinson's disease (PD) and multiple system atrophy (MSA) on mitochondrial and lysosomal functions, and further clarify the role of α-Syn in pathological mechanism of PD and MSA.Methods:Human neuroblastoma cells (SH-SY5Y) were treated with 10 μg/mL α-Syn aggregates incubated with phosphate buffered solution (PBS), and plasma from healthy controls (HCs), PD patients, and MSA patients, respectively; and the untreated SH-SY5Y cells were used as control group. Mito-Tracker Red CMXRos fluorescent staining was used to detect the mitochondrial membrane potential, Western blotting was used to detect the cytochrome C (Cyt C) expression in the mitochondria and cytoplasm, and adenosine triphosphate (ATP) test kit was used to detect the intracellular ATP level. In addition, the lysosomal associated membrane protein-1 (LAMP1) expression was detected by Western blotting, and cathepsin B activity was detected by Magic Red fluorescent staining.Results:(1) Effects of different α-Syn aggregates on mitochondrial function damage: compared with the control group, the PBS, HC, PD and MSA groups had significantly decreased relative fluorescent intensity (Mito-Tracker Red CMXRos staining) in the SH-SY5Y intracellular mitochondria ( P<0.05); the PD group and MSA group had significantly decreased relative fluorescent intensity compared with PBS group and HC group ( P<0.05); the MSA group had significantly decreased relative fluorescent intensity compared with the PD group (41.27%±5.97% vs. 60.05%±4.24%, P<0.05). Compared with the control group, the HC, PD and MSA groups had significantly decreased mitochondrial Cyt C protein expression, while the PBS, HC, PD and MSA groups had significantly increased cytoplasmic Cyt C protein expression ( P<0.05); the PD group and MSA group had significantly decreased mitochondrial Cyt C protein expression compared with the PBS group and HC group ( P<0.05); the MSA group had significantly increased cytoplasmic Cyt C protein expression compared with the PBS group ( P<0.05); the MSA group had significantly decreased mitochondrial Cyt C protein expression (51.50%±4.22%) and increased cytoplasmic Cyt C protein expression (199.40%±6.73%) compared with the PD group (65.52%±2.18% and 174.00%±16.18%, P<0.05). Compared with the control group, the PBS group, HC group, PD group and MSA group had significantly decreased ATP level ( P<0.05); the PD group and MSA group had significantly decreased ATP level compared with the PBS group and HC group ( P<0.05); the MSA group had significantly decreased ATP level (1.81±0.13 nmol/mg) compared with PD group (2.96±0.29 nmol/mg, P<0.05). (2) Effects of different α-Syn aggregates on lysosomal functions: compared with the control group, the PD group and MSA group had significantly decreased LAMP1 protein expression ( P<0.05); the MSA group had significantly decreased LAMP1 protein expression (60.74%±7.94%) compared with the PBS group, HC group, and PD group (79.70%±4.34%, P<0.05). Compared with the control group, the PBS, HC, PD and MSA groups had significantly enhanced relative fluorescent intensity of lysosomal cathepsin B ( P<0.05); the PD group and MSA group had significantly enhanced relative fluorescent intensity of lysosomal cathepsin B compared with PBS group and HC group ( P<0.05); the MSA group had significantly increased relative fluorescent intensity of lysosomal cathepsin B compared with PD group (399.00%±35.54% vs. 305.90%±24.87%, P<0.05). Conclusion:The α-Syn aggregates incubated with plasma from PD and MSA patients have damage to mitochondrial and lysosome functions, especially those incubated with plasma from MSA patients.

Oligoasthenospermia is the primary cause of infertility. However, there are still enormous challenges in the screening of critical candidates and targets of oligoasthenospermia owing to its complex mechanism. In this study, stem cell factor (SCF), c-kit, and transient receptor potential vanilloid 1 (TRPV1) biosensors were successfully established and applied to studying apoptosis and autophagy mechanisms. Interestingly, the detection limit reached 2.787 × 10^-15 g/L, and the quantitative limit reached 1.0 × 10^-13 g/L. Furthermore, biosensors were used to investigate the interplay between autophagy and apoptosis. Schisandrin A is an excellent candidate to form a system with c-kit similar to SCF/c-kit with a detection constant (K_D) of 5.701 × 10^-11 mol/L, whereas it had no affinity for SCF. In addition, it also inhibited autophagy in oligoasthenospermia through antagonizing TRPV1 with a K_D of up to 4.181 × 10^-10 mol/L. In addition, in vivo and in vitro experiments were highly consistent with the biosensor. In summary, high-potency schisandrin A and two potential targets were identified, through which schisandrin A could reverse the apoptosis caused by excessive autophagy during oligoasthenospermia. Our study provides promising insights into the discovery of effective compounds and potential targets via a well-established in vitro-in vivo strategy.