Objective:To explore the static and dynamic improvement effects of dual nerve transfer (hypoglossal nerve-temporal bone trunk end-to-side anastomosis, masseteric nerve-buccal branch end-to-end anastomosis) in the treatment of peripheral facial paralysis after acoustic neuroma resection surgery.Methods:A retrospective analysis was conducted on the clinical data of patients with peripheral facial paralysis after acoustic neuroma surgery who underwent dual nerve transfer surgery in the Department of Plastic Surgery of China-Japan Friendship Hospital from January 2022 to January 2024. The clinical data included standardized photographs and videos before and 1 year after surgery. House-Brackmann (H-B) grading of facial paralysis (Grade Ⅰ to Ⅵ, higher grade indicates worse facial nerve function), electronic facial assessment by computer evaluation (eFACE) (0 to 100 points, higher scores indicate better facial function), facial disability index-physical (FDIP) (-25 to 100 points, higher scores indicate better facial function), facial clinimetric evaluation (FaCE) (0 to 100 points, higher scores indicate better facial function) were used as the effectiveness evaluation indicator, and safety data were collected through patient reports. The measurement data of normal distribution were expressed as Mean±SD and the paired t-test was used. The grade data were analyzed using the Wilcoxon signed-rank test. When conducting correlation tests, the Pearson correlation test was used for normally distributed data. Results:A total of 20 patients were enrolled, including 7 males and 13 females, with an average age of (33.4 ± 6.5) years (22 to 44 years). The duration of facial paralysis was (10.3 ± 4.5) months (1 to 20 months). Before the operation, the H-B grades of all patients were grade Ⅵ. At the 1-year follow-up after the operation, all patients showed significant improvement in facial static tension and smile movement. The H-B grades of the affected sides of all patients improved. Among them, 18 cases decreased from grade Ⅵ to grade Ⅲ, and 2 cases decreased from grade Ⅵ to grade Ⅱ. The differences in H-B grades of the affected sides before and after the operation were statistically significant ( P < 0.01). Compared with those before the operation, the static eFACE scores [(64.55±12.62) points vs. (84.25±9.08) points], dynamic eFACE scores [(34.85±9.31) points vs. (68.70±5.36) points], FDIP scores [(28.50±8.13) points vs. (59.50±5.36) points], and FaCE scores [(23.33±9.23) points vs. (61.92±9.65) points] of the affected sides significantly increased, and the differences were statistically significant (all P<0.01). Correlation analysis showed that there was a negative correlation between age and the postoperative eFACE static score ( r=-0.61, P < 0.01). Five patients complained of mild chewing weakness and bilateral chewing asymmetry, which did not affect their daily lives, no patients had tongue atrophy, tongue extension weakness, speech or swallowing disorders. Conclusion:Dual nerve transfer surgery provides both facial static tension and smile dynamics, and has remarkable clinical efficacy in treating peripheral facial paralysis after acoustic neuroma surgery.

ObjectiveTo explore the molecular mechanism by which gypenoside L （Gyp-L） promotes apoptosis and inhibits ovarian cancer （OC） through the FK506-binding protein （FKBP） prolyl isomerase 8 （FKBP8）/B-cell lymphoma-2 （Bcl-2） axis， with the piR-hsa-2804461 pathway as a breakthrough point. MethodsThe effects of different concentrations of Gyp-L and cis-platinum on the proliferation of OVCAR3 cells were determined by the cell count kit-8 method to identify the appropriate intervention concentration for subsequent experiments. OVCAR3 cells were allocated into blank， low-dose Gyp-L （Gyp-L-L， 50 µmol·L^-1）， high-dose Gyp-L （Gyp-L-H， 100 µmol·L^-1）， and cis-platinum （15 µmol·L^-1） groups. The migration， colony formation， and apoptosis of OVCAR3 cells were detected by the cell scratch assay， colony formation assay， and flow cytometry， respectively. The mRNA levels of piR-hsa-2804461 and FKBP8/Bcl-2 axis-related genes in OVCAR3 cells were determined by Real-time PCR， and the expression levels of FKBP8/Bcl-2 axis-related proteins were determined by simple Western blot. Further， an OVCAR3 cell model with piR-hsa-2804461 knocked out was constructed. The cells were allocated into blank， NC-inhibitor， inhibitor， NC-inhibitor+Gyp-L， and inhibitor+Gyp-L groups. The colony formation of OVCAR3 cells was detected by the colony formation assay. The mRNA levels of piR-hsa-2804461 and FKBP8/Bcl-2 axis-related genes and the expression levels of FKBP8/Bcl-2 axis-related proteins were determined by Real-time PCR and simple Western blotting， respectively. ResultsGyp-L inhibited the migration and proliferation （P<0.01）， promoted the apoptosis （P<0.05）， up-regulated the mRNA level of piR-hsa-2804461 （P<0.05）， and down-regulated the mRNA and protein levels of FKBP8 and Bcl-2 （P<0.05） in OVCAR3 cells. Furthermore， Gyp-L increased the mRNA and protein levels of Bcl-2-associated X protein （Bax）， cysteinyl aspartate-specific proteinase （Caspase）-3， and Caspase-9， which are related to the FKBP8/Bcl-2 axis （P<0.05）. ConclusionGyp-L may promote apoptosis by regulating the piR-hsa-2804461/FKBP8/Bcl-2 axis， thus affecting the occurrence of ovarian cancer.

ObjectiveTo explore the molecular mechanism of gypenoside L （Gyp-L） in the treatment of ovarian cancer （OC） by taking the glycolytic pathway of OC as the key point. MethodsThe proliferation activity of OVCAR3 cells was measured by the cell counting kit-8 （CCK-8） assay to determine the appropriate intervention concentration for subsequent experiments. The cell clone formation assay and the scratch healing assay were employed to assess the proliferation and migration capabilities of OVCAR3 cells. OVCAR3 cells were divided into a blank group， a Gyp-L-L group （low concentration of Gyp-L， 50 µmol·L^-1）， a Gyp-L-H group （high concentration of Gyp-L， 100 µmol·L^-1）， and a cisplatin （15 µmol·L^-1） group. The glucose consumption in OC cells was determined using a kit， and the expression levels of related mRNAs in OVCAR3 cells were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expressions of related proteins were detected by Wes automatic Western blot quantitative analysis. ResultsValidated by the cell scratch assay， the scratch healing rate of OVCAR3 cells was decreased after Gyp-L intervention， reflecting that Gyp-L could significantly inhibit the migration of OVCAR3 cells （P<0.05）. According to the clone formation assay， the cell colony formation ability of the Gyp-L-L group， Gyp-L-H group， and cisplatin group was significantly lower than that of the control group （P<0.05）. In OVCAR3 cells， compared with those in the control group， the levels of glucose consumption and lactate generation in the Gyp-L-L group and Gyp-L-H group were significantly reduced （P<0.05）， indicating that Gyp-L could effectively inhibit the glycolysis level of OC cells. Meanwhile， Gyp-L could suppress the expression levels of glucokinase （GCK）， phosphofructokinase liver （PFKL）， signal transducer and activator of transcription 3 （STAT3）， lactate dehydrogenase D （LDHD）， phosphoglycerate mutase 1 （PGAM1）， mRNAs， and proteins related to the glycolytic pathway in OC cells. ConclusionGyp-L may inhibit the occurrence and development of OC by influencing the GCK-mediated glycolytic pathway.

ObjectiveTo explore the molecular mechanism by which gypenoside L （Gyp-L） promotes apoptosis and inhibits ovarian cancer （OC） through the FK506-binding protein （FKBP） prolyl isomerase 8 （FKBP8）/B-cell lymphoma-2 （Bcl-2） axis， with the piR-hsa-2804461 pathway as a breakthrough point. MethodsThe effects of different concentrations of Gyp-L and cis-platinum on the proliferation of OVCAR3 cells were determined by the cell count kit-8 method to identify the appropriate intervention concentration for subsequent experiments. OVCAR3 cells were allocated into blank， low-dose Gyp-L （Gyp-L-L， 50 µmol·L^-1）， high-dose Gyp-L （Gyp-L-H， 100 µmol·L^-1）， and cis-platinum （15 µmol·L^-1） groups. The migration， colony formation， and apoptosis of OVCAR3 cells were detected by the cell scratch assay， colony formation assay， and flow cytometry， respectively. The mRNA levels of piR-hsa-2804461 and FKBP8/Bcl-2 axis-related genes in OVCAR3 cells were determined by Real-time PCR， and the expression levels of FKBP8/Bcl-2 axis-related proteins were determined by simple Western blot. Further， an OVCAR3 cell model with piR-hsa-2804461 knocked out was constructed. The cells were allocated into blank， NC-inhibitor， inhibitor， NC-inhibitor+Gyp-L， and inhibitor+Gyp-L groups. The colony formation of OVCAR3 cells was detected by the colony formation assay. The mRNA levels of piR-hsa-2804461 and FKBP8/Bcl-2 axis-related genes and the expression levels of FKBP8/Bcl-2 axis-related proteins were determined by Real-time PCR and simple Western blotting， respectively. ResultsGyp-L inhibited the migration and proliferation （P<0.01）， promoted the apoptosis （P<0.05）， up-regulated the mRNA level of piR-hsa-2804461 （P<0.05）， and down-regulated the mRNA and protein levels of FKBP8 and Bcl-2 （P<0.05） in OVCAR3 cells. Furthermore， Gyp-L increased the mRNA and protein levels of Bcl-2-associated X protein （Bax）， cysteinyl aspartate-specific proteinase （Caspase）-3， and Caspase-9， which are related to the FKBP8/Bcl-2 axis （P<0.05）. ConclusionGyp-L may promote apoptosis by regulating the piR-hsa-2804461/FKBP8/Bcl-2 axis， thus affecting the occurrence of ovarian cancer.

ObjectiveTo explore the molecular mechanism of gypenoside L （Gyp-L） in the treatment of ovarian cancer （OC） by taking the glycolytic pathway of OC as the key point. MethodsThe proliferation activity of OVCAR3 cells was measured by the cell counting kit-8 （CCK-8） assay to determine the appropriate intervention concentration for subsequent experiments. The cell clone formation assay and the scratch healing assay were employed to assess the proliferation and migration capabilities of OVCAR3 cells. OVCAR3 cells were divided into a blank group， a Gyp-L-L group （low concentration of Gyp-L， 50 µmol·L^-1）， a Gyp-L-H group （high concentration of Gyp-L， 100 µmol·L^-1）， and a cisplatin （15 µmol·L^-1） group. The glucose consumption in OC cells was determined using a kit， and the expression levels of related mRNAs in OVCAR3 cells were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expressions of related proteins were detected by Wes automatic Western blot quantitative analysis. ResultsValidated by the cell scratch assay， the scratch healing rate of OVCAR3 cells was decreased after Gyp-L intervention， reflecting that Gyp-L could significantly inhibit the migration of OVCAR3 cells （P<0.05）. According to the clone formation assay， the cell colony formation ability of the Gyp-L-L group， Gyp-L-H group， and cisplatin group was significantly lower than that of the control group （P<0.05）. In OVCAR3 cells， compared with those in the control group， the levels of glucose consumption and lactate generation in the Gyp-L-L group and Gyp-L-H group were significantly reduced （P<0.05）， indicating that Gyp-L could effectively inhibit the glycolysis level of OC cells. Meanwhile， Gyp-L could suppress the expression levels of glucokinase （GCK）， phosphofructokinase liver （PFKL）， signal transducer and activator of transcription 3 （STAT3）， lactate dehydrogenase D （LDHD）， phosphoglycerate mutase 1 （PGAM1）， mRNAs， and proteins related to the glycolytic pathway in OC cells. ConclusionGyp-L may inhibit the occurrence and development of OC by influencing the GCK-mediated glycolytic pathway.

Objective:To explore the static and dynamic improvement effects of dual nerve transfer (hypoglossal nerve-temporal bone trunk end-to-side anastomosis, masseteric nerve-buccal branch end-to-end anastomosis) in the treatment of peripheral facial paralysis after acoustic neuroma resection surgery.Methods:A retrospective analysis was conducted on the clinical data of patients with peripheral facial paralysis after acoustic neuroma surgery who underwent dual nerve transfer surgery in the Department of Plastic Surgery of China-Japan Friendship Hospital from January 2022 to January 2024. The clinical data included standardized photographs and videos before and 1 year after surgery. House-Brackmann (H-B) grading of facial paralysis (Grade Ⅰ to Ⅵ, higher grade indicates worse facial nerve function), electronic facial assessment by computer evaluation (eFACE) (0 to 100 points, higher scores indicate better facial function), facial disability index-physical (FDIP) (-25 to 100 points, higher scores indicate better facial function), facial clinimetric evaluation (FaCE) (0 to 100 points, higher scores indicate better facial function) were used as the effectiveness evaluation indicator, and safety data were collected through patient reports. The measurement data of normal distribution were expressed as Mean±SD and the paired t-test was used. The grade data were analyzed using the Wilcoxon signed-rank test. When conducting correlation tests, the Pearson correlation test was used for normally distributed data. Results:A total of 20 patients were enrolled, including 7 males and 13 females, with an average age of (33.4 ± 6.5) years (22 to 44 years). The duration of facial paralysis was (10.3 ± 4.5) months (1 to 20 months). Before the operation, the H-B grades of all patients were grade Ⅵ. At the 1-year follow-up after the operation, all patients showed significant improvement in facial static tension and smile movement. The H-B grades of the affected sides of all patients improved. Among them, 18 cases decreased from grade Ⅵ to grade Ⅲ, and 2 cases decreased from grade Ⅵ to grade Ⅱ. The differences in H-B grades of the affected sides before and after the operation were statistically significant ( P < 0.01). Compared with those before the operation, the static eFACE scores [(64.55±12.62) points vs. (84.25±9.08) points], dynamic eFACE scores [(34.85±9.31) points vs. (68.70±5.36) points], FDIP scores [(28.50±8.13) points vs. (59.50±5.36) points], and FaCE scores [(23.33±9.23) points vs. (61.92±9.65) points] of the affected sides significantly increased, and the differences were statistically significant (all P<0.01). Correlation analysis showed that there was a negative correlation between age and the postoperative eFACE static score ( r=-0.61, P < 0.01). Five patients complained of mild chewing weakness and bilateral chewing asymmetry, which did not affect their daily lives, no patients had tongue atrophy, tongue extension weakness, speech or swallowing disorders. Conclusion:Dual nerve transfer surgery provides both facial static tension and smile dynamics, and has remarkable clinical efficacy in treating peripheral facial paralysis after acoustic neuroma surgery.

AIM:Using bioinformatics analysis and experiment validation to explore the differential expres-sion genes related to abnormal lipid metabolism in hepatocellular carcinoma(HCC)and the molecular mechanism of pachymaran affecting pyroptosis through squalene epoxidase(SQLE)/nucleotide-binding oligomerization domain-like re-ceptor protein 3(NLRP3)/gasdermin D(GSDMD)signaling pathway.METHODS:(1)The GEO,GSEA,DAVID,STRING and GEPIA databases were employed to screen abnormal lipid metabolism-related differentially expressed genes in HCC.(2)The tumor tissues from HCC patients(n=9)were collected to verify the differential expression of SQLE.(3)The inhibitory effect of pachymaran on the viability of human HCC cell line HepG2 was measured by CCK-8 assay.(4)The HepG2 cells were divided into control group and pachymaran(800 mg/L)group.The cell migration was analyzed by wound-healing assay,and RT-qPCR was used to measure SQLE mRNA expression.(5)The HepG2 cells with overexpres-sion of SQLE(OE-SQLE)were divided into 5 groups as follows:control group,overexpression negative control(OE-NC)group,OE-SQLE group,OE-NC+pachymaran group,and OE-SQLE+pachymaran group.The mRNA and protein expres-sion levels of SQLE and pyroptosis-related factors were determined by RT-qPCR and Western blot.Colorimetric method and ELISA were used to measure lactate dehydrogenase(LDH),interleukin-1β(IL-1β)and IL-18 levels.The necrosis of HepG2 cells was analyzed by flow cytometry.RESULTS:The SQLE gene was screened through bioinformatics analysis,and its mRNA expression was significantly increased in tumor tissues from HCC patients(P＜0.01).In cell experiments,treatment with 800 mg/L pachymaran for 48 h had a significant inhibitory effect on HepG2 cell viability,and the expres-sion of SQLE mRNA was reduced(P＜0.01).After overexpression of SQLE,the mRNA and protein levels of pyroptosis-re-lated factors,necrotic rate,and LDH,IL-1β and IL-18 levels were significantly decreased(P＜0.05).After treatment with pachymaran,the above indicators were significantly increased(P＜0.05).CONCLUSION:The SQLE is abnormal-ly highly expressed in HCC,and pachymaran can affect the growth of HCC cells by activating the NLRP3/GSDMD pyropto-sis pathway through SQLE.

OBJECTIVETo explore the expression of KIF18A gene protein in gastric cancer tissues and its association with the prognosis of patients.

METHODSTwenty fresh paired gastric cancer specimens and adjacent normal mucosa(at least 5 cm from the edge of tumor) from 20 gastric cancer patients undergoing operation in Department of General Surgery at the First Affiliated Hosptial of Anhui Medical University between March 2015 and July 2015 were collected. Real-time PCR was used to examine KIF18A mRNA expression in above specimens. Meanwhile, paraffin embedded cancer tissue samples from 129 gastric cancer patients undergoing operation and 23 samples of randomly selected normal gastric tissue(adjacent non-cancer tissue) were collected to establish the microarray. Immunohistochemistry method was applied to detect the KIF18A protein expression in the microarray after confirmation by pathologists. Association of KIF18A expression with clinicopathological features in gastric cancer patients was evaluated. Cox proportional hazard model was used to identify prognostic risk factors.

RESULTSAmong 20 fresh paired gastric cancer specimens, mRNA expression of KIF18A in 16 specimens was obviously lower than that in adjacent normal tissues. The positive rate of KIF18A protein expression in gastric cancer tissues was significantly lower than that in normal gastric tissues in microarray[45.0%(58/129) vs. 69.6%(16/23), P=0.041]. KIF18A protein expression was significantly associated with invasion depth (P=0.008) and TNM staging (P=0.032). The median overall survival of all the 129 patients was 44.0(95% CI: 39.78-49.24) months. The three-year survival rates of patients with high and low KIF18A expression were 67.2% and 36.6% respectively(P=0.020). Cox regression analysis showed that KIF18A expression was an independent protective factor of the prognosis of gastric cancer patients (HR=0.570, 95% CI:0.335 to 0.970).

CONCLUSIONSKIF18A expression is down-regulated in gastric cancer tissue, which may play a critical role in gastric cancer carcinogenesis. Lower expression of KIF18A is associated with poor prognosis of gastric cancer patients. KIF18A may be a potential prognostic marker of gastric cancer.

Biomarkers, Tumor ; metabolism ; Humans ; Immunohistochemistry ; Kinesin ; metabolism ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Real-Time Polymerase Chain Reaction ; Regression Analysis ; Stomach Neoplasms ; diagnosis ; metabolism ; Survival Rate