Objective:To investigate the effect of the antioxidant Tempol on the skin depigmentation of a vitiligo-like mouse model induced by the combination of the endogenous reactive oxygen species (ROS) producer AAPH and tyrosinase-related protein 2 (TRP2) -180 nonapeptide.Methods:A vitiligo-like skin depigmentation model was established by immunizing mice with injections of a TRP2-180 nonapeptide mixture into the foot pads twice and into the tails twice, with the injection interval being 1 week. After the first injection, 12 immune-induced mouse models of vitiligo were randomly divided into 4 groups (3 mice per group) : a model group, an AAPH group, a Tempol group, and a combined treatment group; additionally, 3 untreated mice injected with an ovalbumin (OVA) 257-264 peptide served as a sham control group. Mice in the AAPH group, the Tempol group, and the combined treatment group were subcutaneously injected with AAPH into the tails, intraperitoneally injected with Tempol, and received the above both treatments, respectively, while mice in the model group and the sham control group received phosphate-buffered saline injections into the tail and/or abdomen. Drug interventions were carried out 3 times per week for 3 consecutive weeks. Six weeks after the last immunization, mice were sacrificed. The area of depigmented macules on the tail was measured using a point-counting method, X-gal staining and double immunofluorescence staining were performed to assess the distribution and number of melanocytes, mast cells, and CD8 + T cells in depigmented macules on the tail. HaCaT cells were in vitro co-cultured with AAPH and/or Tempol, and a conventional culture group served as the control. Cellular ROS levels were measured by dichlorofluorescin diacetate labeling and flow cytometry; Western blot analysis was performed to determine the expression of matrix metalloproteinase 9 (MMP9) and stem cell factor (SCF) in cell lysates, and to detect soluble SCF levels in the culture supernatant. Comparisons among multiple groups were conducted using one-way analysis of variance, and multiple comparisons were performed using least significant difference- t test. Results:Depigmented macules were observed on the tails of mice in all groups except the sham control group. The area of depigmented macules was significantly larger in the AAPH group (7.27 ± 0.31 cm 2) than in the model group and combined treatment group (3.53 ± 0.21 cm 2, 4.07 ± 0.40 cm 2; t = 13.48, 11.56, respectively, both P < 0.001), while there was no significant difference between the Tempol group (3.30 ± 0.40 cm 2) and the model group ( P = 0.424). X-gal staining and double immunofluorescence staining showed that the number of melanocytes in the normal skin around the depigmented macules was significantly lower in the AAPH group and the combined treatment group than in the model group ( t = 6.31, 5.16, respectively, both P < 0.001), and no significant difference was observed between the AAPH group and the combined treatment group ( P = 0.516). The numbers of CD8 + T cells and mast cells were significantly higher in the AAPH group than in the model group and the combined treatment group (all P < 0.001). The numbers of the 3 types of cells mentioned above in the Tempol group did not differ from those in the model group (all P > 0.05). The ROS levels in HaCaT cells in the AAPH group were the highest, and significantly higher than those in the control group and the combined treatment group (both P < 0.001). Western blot analysis showed that the MMP9 level in the cell lysates and soluble SCF level in the culture supernatant were significantly higher in the AAPH group than in the control group and the combined treatment group (all P < 0.05) ; no significant difference was observed in the membrane-bound SCF level in cell lysates among the groups ( F = 0.06, P = 0.977) . Conclusion:The antioxidant Tempol could inhibit the formation of skin depigmented macules in vitiligo-like mouse models under AAPH-induced oxidative stress.

Objective:To investigate effects of rutin on ultraviolet irradiation (UVR) -induced human dermal fibroblast (FB) senescence and melanogenesis in mouse ear skin.Methods:The third- to fifth-passage FBs were divided into 4 groups: a blank control group, a UVR group, a rutin group, and a combined treatment group. In the UVR group, FBs were irradiated using an ultraviolet irradiator at a single dose of 0.6 J/cm 2 UVA combined with 0.03 J/cm 2 UVB once daily for 5 consecutive days; FBs in the rutin group were cultured in Dulbecco's modified Eagle medium containing 50 μmol/L rutin for 5 days; the combined treatment group received both UVR and the treatment with 50 μmol/L rutin for 5 days; the blank control group underwent no treatment. β-Galactosidase staining was performed to assess the senescence of FBs, real-time quantitative PCR (qPCR) to measure the telomere length in FBs, and Western blot analysis to detect the expression levels of stem cell factor (SCF) in FB cell lysates and culture supernatants. FB culture supernatants were collected from each group, and mixed with M254 medium at a ratio of 3∶1 to prepare conditioned medium, which was then used to treat PIG1 melanocytes for 24 hours. Western blot analysis was conducted to determine the tyrosinase (TYR) expression in PIG1 melanocytes in each group, while the 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay was applied to assess the proliferative activity of PIG1 cells in each group. Ten Dct-LacZ transgenic mice were divided into a control group and a UVR group. For each mouse, 5% rutin-containing cream was topically applied to the right ear after UVR, while the left ear treated with the cream base alone served as a control. Skin biopsies were performed after 4 weeks, followed by X-gal staining and Avidin/fluorescein isothiocyanate (FITC) staining to count the numbers of melanocytes and mast cells in mouse ear skin. Results:In the UVR group, the number of senescent FBs (25.67 ± 2.89), relative protein expression levels of SCF (1.95 ± 0.22), and relative levels of SCF in the cell culture supernatant (1.52 ± 0.34) were all significantly higher than those in the blank control group (5.67 ± 1.56, 0.95 ± 0.11, 1.01 ± 0.31, respectively), while these indicators were significantly lower in the combined treatment group (12.00 ± 1.63, 1.32 ± 0.19, 1.15 ± 0.32, respectively) than in the UVR group (all P < 0.05). The relative telomere length in FBs was significantly shorter in the UVR group (0.49 ± 0.12) than in the blank control group (0.94 ± 0.11; LSD- t = 3.15, P = 0.021), but significantly longer in the combined treatment group (0.81 ± 0.13) than in the UVR group (LSD- t = 4.30, P = 0.034). After the treatment with FB conditioned medium, the relative expression level of TYR in PIG1 melanocytes and the number of EdU-positive cells were significantly higher in the UVR group (2.54 ± 0.21, 33.54 ± 3.21, respectively) than in the blank control group (0.97 ± 0.19, 21.45 ± 2.51, respectively; both P < 0.001), but significantly lower in the combined treatment group (1.63 ± 0.12, 18.54 ± 3.87, respectively) than in the UVR group (both P < 0.001). X-gal staining and Avidin/FITC staining showed that the numbers of melanocytes and mast cells in the mouse left ear skin in the UVR group (5.00 ± 1.22, 98.60 ± 8.47, respectively) were significantly higher than those in the mouse left ear skin in the control group (1.80 ± 0.45, 53.80 ± 5.76, respectively) and those in the mouse right ear skin treated with the rutin-containing cream in the UVR group (2.80 ± 0.45, 69.60 ± 8.89, respectively) (all P < 0.05) . Conclusion:Rutin may inhibit UVR-induced skin melanogenesis by suppressing the senescence of dermal FBs and paracrine secretion of SCF.

Objective:By combining image memory with educational games, electrocardiogram (ECG) images were integrated with card games to simplify complex ECG knowledge and visualize abstract concepts. This approach stimulated students' motivation to learn within the game, thereby enhancing their cognitive abilities and aiding them in mastering ECG-related knowledge.Methods:Through literature search and brainstorming, ECG learning flash cards and game process were designed. Using the convenience sampling method, 55 medical students from a university were selected to conduct the self-designed questionnaire before and after using ECG flash cards to evaluate the efficacy of flash cards in enhancing ECG learning outcomes. A descriptive qualitative study was conducted on the satisfaction with the flash cards.Results:Before the intervention, there was no significant difference in the score of students from different majors. For students who had studied ECG within 1 month and 6 months, the difference in correct recognition rate between them was relatively small, which were 57.50% and 54.51%, respectively. For students who had not studied ECG, the correct recognition rate after training was only 18.06%. After the intervention, 42 valid questionnaires were collected. The overall correct rate of ECG recognition increased by 17.86%. The correct recognition rate of ventricular fibrillation increased significantly from 30.91% to 64.29%; the correct recognition rate of supraventricular tachycardia was improved only by 5.50%. The correct ECG recognition rate was significantly higher after the intervention than before the intervention ( P<0.01). The results of qualitative interviews indicated that the ECG game cards were interesting, practical, and generalizable. However, there was room for improvement in the distinctiveness of key graphics, the richness of information on cards, and the diversity of rule design. Conclusions:ECG recognition is hard to master for beginners. The use of ECG flash cards simplifies the process of ECG recognition and stimulates students' initiative in learning, thus improving the overall correct ECG recognition rate.

Objective:To investigate the effect of the antioxidant Tempol on the skin depigmentation of a vitiligo-like mouse model induced by the combination of the endogenous reactive oxygen species (ROS) producer AAPH and tyrosinase-related protein 2 (TRP2) -180 nonapeptide.Methods:A vitiligo-like skin depigmentation model was established by immunizing mice with injections of a TRP2-180 nonapeptide mixture into the foot pads twice and into the tails twice, with the injection interval being 1 week. After the first injection, 12 immune-induced mouse models of vitiligo were randomly divided into 4 groups (3 mice per group) : a model group, an AAPH group, a Tempol group, and a combined treatment group; additionally, 3 untreated mice injected with an ovalbumin (OVA) 257-264 peptide served as a sham control group. Mice in the AAPH group, the Tempol group, and the combined treatment group were subcutaneously injected with AAPH into the tails, intraperitoneally injected with Tempol, and received the above both treatments, respectively, while mice in the model group and the sham control group received phosphate-buffered saline injections into the tail and/or abdomen. Drug interventions were carried out 3 times per week for 3 consecutive weeks. Six weeks after the last immunization, mice were sacrificed. The area of depigmented macules on the tail was measured using a point-counting method, X-gal staining and double immunofluorescence staining were performed to assess the distribution and number of melanocytes, mast cells, and CD8 + T cells in depigmented macules on the tail. HaCaT cells were in vitro co-cultured with AAPH and/or Tempol, and a conventional culture group served as the control. Cellular ROS levels were measured by dichlorofluorescin diacetate labeling and flow cytometry; Western blot analysis was performed to determine the expression of matrix metalloproteinase 9 (MMP9) and stem cell factor (SCF) in cell lysates, and to detect soluble SCF levels in the culture supernatant. Comparisons among multiple groups were conducted using one-way analysis of variance, and multiple comparisons were performed using least significant difference- t test. Results:Depigmented macules were observed on the tails of mice in all groups except the sham control group. The area of depigmented macules was significantly larger in the AAPH group (7.27 ± 0.31 cm 2) than in the model group and combined treatment group (3.53 ± 0.21 cm 2, 4.07 ± 0.40 cm 2; t = 13.48, 11.56, respectively, both P < 0.001), while there was no significant difference between the Tempol group (3.30 ± 0.40 cm 2) and the model group ( P = 0.424). X-gal staining and double immunofluorescence staining showed that the number of melanocytes in the normal skin around the depigmented macules was significantly lower in the AAPH group and the combined treatment group than in the model group ( t = 6.31, 5.16, respectively, both P < 0.001), and no significant difference was observed between the AAPH group and the combined treatment group ( P = 0.516). The numbers of CD8 + T cells and mast cells were significantly higher in the AAPH group than in the model group and the combined treatment group (all P < 0.001). The numbers of the 3 types of cells mentioned above in the Tempol group did not differ from those in the model group (all P > 0.05). The ROS levels in HaCaT cells in the AAPH group were the highest, and significantly higher than those in the control group and the combined treatment group (both P < 0.001). Western blot analysis showed that the MMP9 level in the cell lysates and soluble SCF level in the culture supernatant were significantly higher in the AAPH group than in the control group and the combined treatment group (all P < 0.05) ; no significant difference was observed in the membrane-bound SCF level in cell lysates among the groups ( F = 0.06, P = 0.977) . Conclusion:The antioxidant Tempol could inhibit the formation of skin depigmented macules in vitiligo-like mouse models under AAPH-induced oxidative stress.

Objective:To investigate effects of rutin on ultraviolet irradiation (UVR) -induced human dermal fibroblast (FB) senescence and melanogenesis in mouse ear skin.Methods:The third- to fifth-passage FBs were divided into 4 groups: a blank control group, a UVR group, a rutin group, and a combined treatment group. In the UVR group, FBs were irradiated using an ultraviolet irradiator at a single dose of 0.6 J/cm 2 UVA combined with 0.03 J/cm 2 UVB once daily for 5 consecutive days; FBs in the rutin group were cultured in Dulbecco's modified Eagle medium containing 50 μmol/L rutin for 5 days; the combined treatment group received both UVR and the treatment with 50 μmol/L rutin for 5 days; the blank control group underwent no treatment. β-Galactosidase staining was performed to assess the senescence of FBs, real-time quantitative PCR (qPCR) to measure the telomere length in FBs, and Western blot analysis to detect the expression levels of stem cell factor (SCF) in FB cell lysates and culture supernatants. FB culture supernatants were collected from each group, and mixed with M254 medium at a ratio of 3∶1 to prepare conditioned medium, which was then used to treat PIG1 melanocytes for 24 hours. Western blot analysis was conducted to determine the tyrosinase (TYR) expression in PIG1 melanocytes in each group, while the 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay was applied to assess the proliferative activity of PIG1 cells in each group. Ten Dct-LacZ transgenic mice were divided into a control group and a UVR group. For each mouse, 5% rutin-containing cream was topically applied to the right ear after UVR, while the left ear treated with the cream base alone served as a control. Skin biopsies were performed after 4 weeks, followed by X-gal staining and Avidin/fluorescein isothiocyanate (FITC) staining to count the numbers of melanocytes and mast cells in mouse ear skin. Results:In the UVR group, the number of senescent FBs (25.67 ± 2.89), relative protein expression levels of SCF (1.95 ± 0.22), and relative levels of SCF in the cell culture supernatant (1.52 ± 0.34) were all significantly higher than those in the blank control group (5.67 ± 1.56, 0.95 ± 0.11, 1.01 ± 0.31, respectively), while these indicators were significantly lower in the combined treatment group (12.00 ± 1.63, 1.32 ± 0.19, 1.15 ± 0.32, respectively) than in the UVR group (all P < 0.05). The relative telomere length in FBs was significantly shorter in the UVR group (0.49 ± 0.12) than in the blank control group (0.94 ± 0.11; LSD- t = 3.15, P = 0.021), but significantly longer in the combined treatment group (0.81 ± 0.13) than in the UVR group (LSD- t = 4.30, P = 0.034). After the treatment with FB conditioned medium, the relative expression level of TYR in PIG1 melanocytes and the number of EdU-positive cells were significantly higher in the UVR group (2.54 ± 0.21, 33.54 ± 3.21, respectively) than in the blank control group (0.97 ± 0.19, 21.45 ± 2.51, respectively; both P < 0.001), but significantly lower in the combined treatment group (1.63 ± 0.12, 18.54 ± 3.87, respectively) than in the UVR group (both P < 0.001). X-gal staining and Avidin/FITC staining showed that the numbers of melanocytes and mast cells in the mouse left ear skin in the UVR group (5.00 ± 1.22, 98.60 ± 8.47, respectively) were significantly higher than those in the mouse left ear skin in the control group (1.80 ± 0.45, 53.80 ± 5.76, respectively) and those in the mouse right ear skin treated with the rutin-containing cream in the UVR group (2.80 ± 0.45, 69.60 ± 8.89, respectively) (all P < 0.05) . Conclusion:Rutin may inhibit UVR-induced skin melanogenesis by suppressing the senescence of dermal FBs and paracrine secretion of SCF.

Objective:By combining image memory with educational games, electrocardiogram (ECG) images were integrated with card games to simplify complex ECG knowledge and visualize abstract concepts. This approach stimulated students' motivation to learn within the game, thereby enhancing their cognitive abilities and aiding them in mastering ECG-related knowledge.Methods:Through literature search and brainstorming, ECG learning flash cards and game process were designed. Using the convenience sampling method, 55 medical students from a university were selected to conduct the self-designed questionnaire before and after using ECG flash cards to evaluate the efficacy of flash cards in enhancing ECG learning outcomes. A descriptive qualitative study was conducted on the satisfaction with the flash cards.Results:Before the intervention, there was no significant difference in the score of students from different majors. For students who had studied ECG within 1 month and 6 months, the difference in correct recognition rate between them was relatively small, which were 57.50% and 54.51%, respectively. For students who had not studied ECG, the correct recognition rate after training was only 18.06%. After the intervention, 42 valid questionnaires were collected. The overall correct rate of ECG recognition increased by 17.86%. The correct recognition rate of ventricular fibrillation increased significantly from 30.91% to 64.29%; the correct recognition rate of supraventricular tachycardia was improved only by 5.50%. The correct ECG recognition rate was significantly higher after the intervention than before the intervention ( P<0.01). The results of qualitative interviews indicated that the ECG game cards were interesting, practical, and generalizable. However, there was room for improvement in the distinctiveness of key graphics, the richness of information on cards, and the diversity of rule design. Conclusions:ECG recognition is hard to master for beginners. The use of ECG flash cards simplifies the process of ECG recognition and stimulates students' initiative in learning, thus improving the overall correct ECG recognition rate.

Objective:To investigate the quantification assessment of segmental volume after thigh liposuction utilizing three-dimensional(3D) scanning technology.Methods:This retrospective study was performed with the analysis of 3D scanning images of patients who had undergone bilateral thigh liposuction in Body Contouring & Fat Grafting Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2018 to September 2022. Preoperative and postoperative 3D scanning were performed to build visual 3D models of bilateral thighs. From top to the bottom, bilateral thighs were segmented into seven sections with a space of 5 cm in acquired 3D model. Certain measurements and calculation of preoperative and postoperative volume and volume change rate of the overall thigh and each segment were conducted, to validate the efficiency of liposuction (EOL). Additionally, EOL of each thigh segment was computed and the symmetry of bilateral thighs was analyzed before and after surgery. The volume differences were computed using the paired Wilcoxon rank-sum test, and intraclass correlation coefficient (ICC) was exerted to assess the symmetry of bilateral thighs before and after surgery.Results:A total of 36 female patients were included in the study, with an age range of 18 to 49 years and a mean age of (28.6±7.6) years. Follow-up duration ranged from 23 to 1 133 days postoperatively, with a mean follow-up period of 274.7 days. The results of 3D scanning measurements indicated significant changes ( P<0.01) in both the overall volume of the thigh and the volumes of each segment before and after surgery. The highest volume change rate and EOL were observed in the uppermost segment, and the volumetric change rate and EOL exhibited a descending trend across the segments of the thigh from the uppermost to the lowermost segments. The ICC of the volume of each segment consistently surpassed 0.950 whether preoperatively or postoperatively, indicating a high level of symmetry between the bilateral thighs, and the ICC of overall volume showed a notable increase from 0.992 preoperatively to 0.997 postoperatively. Conclusion:3D scanning technology can be exerted to quantify the volume changes before and after thigh liposuction. This study provided quantitative and objective evidence to confirm the efficacy of thigh liposuction procedure, elucidating that the most significant liposuction effects observed in the uppermost segment of the thigh. Moreover, postoperative assessments reveal a further enhancement in bilateral thigh symmetry.

Obesity and related metabolic syndromes have been recognized as important disease risks,in which the role of adipokines cannot be ignored.Adiponectin(ADP)is one of the key adipokines with various beneficial effects,including improving glucose and lipid metabolism,enhancing insulin sensitivity,reducing oxidative stress and inflammation,promoting ceramides degradation,and stimulating adipose tissue vascularity.Based on those,it can serve as a positive regulator in many metabolic syndromes,such as type 2 diabetes(T2D),cardiovascular diseases,non-alcoholic fatty liver disease(NAFLD),sarcopenia,neurodegenerative diseases,and certain cancers.Therefore,a promising therapeutic approach for treating various metabolic diseases may involve elevating ADP levels or activating ADP receptors.The modulation of ADP genes,multimerization,and secretion covers the main processes of ADP generation,providing a comprehensive orientation for the development of more appropriate therapeutic strategies.In order to have a deeper understanding of ADP,this paper will provide an all-encompassing review of ADP.

Polycystic ovary syndrome（PCOS） is a common reproductive endocrine and metabolic disorder， affecting about 6%-10% of women of childbearing age worldwide， and has always been a major challenge in clinical treatment. Guizhi Fulingwan， derived from the Synopsis of the Golden Chamber（《金匮要略》）， has the effect of promoting blood circulation， removing blood stasis， and alleviating blockages. In recent years， many studies have found that it has a good therapeutic effect on PCOS. By sorting out the research literature on the mechanism of Guizhi Fulingwan treating PCOS， it is found that this formula can decrease the autophagy and apoptosis of ovarian granulosa cells， correct endocrine hormone disorder， improve the inflammatory environment， alleviate oxidative stress. Moreover， it regulates phosphatidylinositide 3-kinase（PI3K）/protein kinase B（Akt）/mammalian target of rapamycin（mTOR）， lncRNA H19/miR-29b-3p， nuclear factor E₂-related factor 2（Nrf2）/heme oxygenase-1（HO-1）， PI3K/Akt/nuclear factor-κB（NF-κB）， and other signaling pathways. And also effects the regulator expression of matrix metallopeptidase-9（MMP-9）， matrix metalloproteinase inhibitor-1（TIMP-1）， cytochrome P45019A1 enzyme（CYP19A1）， glucose transporter protein 4（GLUT4）， regulated on activation normal T cell expressed and secreted（RANTES）， monocyte chemoattractant protein-1（MCP-1）. Thus， Guizhi Fulingwan is confirmed to treat PCOS based on these molecular mechanisms. Besides， the main active ingredients of the formula such as cinnamic aldehyde， （+）-catechin， stigmasterol， also have the effect of anti-oxidant， anti-inflammatory， regulating mitochondrial function， regulating endocrine， and regulating metabolism. According to the summary of clinical literature on Guizhi Fulingwan， it is found that the use of the formula alone or in combination with other drugs can significantly alleviate the clinical symptoms and complications of PCOS. Thus， it is safe with no adverse reactions in long-term use， which is worthily further promoted in clinical application. This paper comprehensively analyzed the current research status of Guizhi Fulingwan for the treatment of PCOS through the mechanism and clinical studies， summarized the limitations in current research， and put forward suggestions， aiming to provide the strong literature support for the future in-depth research on and clinical application of the formula.

Objective:To investigate the quantification assessment of segmental volume after thigh liposuction utilizing three-dimensional(3D) scanning technology.Methods:This retrospective study was performed with the analysis of 3D scanning images of patients who had undergone bilateral thigh liposuction in Body Contouring & Fat Grafting Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2018 to September 2022. Preoperative and postoperative 3D scanning were performed to build visual 3D models of bilateral thighs. From top to the bottom, bilateral thighs were segmented into seven sections with a space of 5 cm in acquired 3D model. Certain measurements and calculation of preoperative and postoperative volume and volume change rate of the overall thigh and each segment were conducted, to validate the efficiency of liposuction (EOL). Additionally, EOL of each thigh segment was computed and the symmetry of bilateral thighs was analyzed before and after surgery. The volume differences were computed using the paired Wilcoxon rank-sum test, and intraclass correlation coefficient (ICC) was exerted to assess the symmetry of bilateral thighs before and after surgery.Results:A total of 36 female patients were included in the study, with an age range of 18 to 49 years and a mean age of (28.6±7.6) years. Follow-up duration ranged from 23 to 1 133 days postoperatively, with a mean follow-up period of 274.7 days. The results of 3D scanning measurements indicated significant changes ( P<0.01) in both the overall volume of the thigh and the volumes of each segment before and after surgery. The highest volume change rate and EOL were observed in the uppermost segment, and the volumetric change rate and EOL exhibited a descending trend across the segments of the thigh from the uppermost to the lowermost segments. The ICC of the volume of each segment consistently surpassed 0.950 whether preoperatively or postoperatively, indicating a high level of symmetry between the bilateral thighs, and the ICC of overall volume showed a notable increase from 0.992 preoperatively to 0.997 postoperatively. Conclusion:3D scanning technology can be exerted to quantify the volume changes before and after thigh liposuction. This study provided quantitative and objective evidence to confirm the efficacy of thigh liposuction procedure, elucidating that the most significant liposuction effects observed in the uppermost segment of the thigh. Moreover, postoperative assessments reveal a further enhancement in bilateral thigh symmetry.