Objective This study aimed to investigate whether photodynamic therapy(PDT)with methylene blue(MB)as a photosensitizer can induce ferroptosis in melanoma cells and its potential mechanisms.Methods The paraffin sections of malignant melanoma patients(5 cases)and malignant melanoma patients with lymph node metastasis(5 cases)were collected.Prussian blue,TUNEL and immunohistochemical staining were performed to compare the degree of ferroptosis between the two groups.CCK-8 assay and scratch assay were used to detect the cytotoxic effect of methylene blue and methylene-blue-mediated photodynamic therapy on B16F10 under different treatment conditions and the effect of cell migration ability.The expression of ferroptosis-related proteins(glutathione peroxidase 4,GPX4、solute carrier family 7 member 11,SLC7A11)in B16F10 cells was detected by Western blot.The expression levels of intracellular malondialdehyde(MDA)and GSH/GSSG were detected by microplate reader.The expression levels of reactive oxygen species(ROS),Fe2+and lipid peroxide(LPO)in B16F10 cells were detected by flow cytometry.Results Compared with melanoma cells transferred to lymph nodes,the non-metastatic group had a higher degree of ferroptosis and a lower expression level of SLC7A11.The cytotoxicity of MB and MB-PDT on B16F10 was dose-dependent(P＜0.000 1).The results of scratch assay showed that MB-PDT inhibited the migration ability of B16F10 cells(P＜0.001).Compared with the control group,after MB-PDT treatment,the expression levels of GPX4,SLC7A11 and GSH/GSSG were decreased(P＜0.05),and the expression levels of ROS,Fe2+,LPO and MDA were increased(P＜0.05)in B16F10 cells.Pretreatment with Ferrostatin-1,an inhibitor of ferroptosis,inhibited MB-PDT-induced cytotoxicity and ferroptosis in B16F10 cells(P＜0.05).Conclusion MB-PDT can induce ferroptosis of melanoma cells by inhibiting SLC7A11 expression,thereby inhibiting the progression of melanoma.

Objective This study aimed to investigate whether photodynamic therapy(PDT)with methylene blue(MB)as a photosensitizer can induce ferroptosis in melanoma cells and its potential mechanisms.Methods The paraffin sections of malignant melanoma patients(5 cases)and malignant melanoma patients with lymph node metastasis(5 cases)were collected.Prussian blue,TUNEL and immunohistochemical staining were performed to compare the degree of ferroptosis between the two groups.CCK-8 assay and scratch assay were used to detect the cytotoxic effect of methylene blue and methylene-blue-mediated photodynamic therapy on B16F10 under different treatment conditions and the effect of cell migration ability.The expression of ferroptosis-related proteins(glutathione peroxidase 4,GPX4、solute carrier family 7 member 11,SLC7A11)in B16F10 cells was detected by Western blot.The expression levels of intracellular malondialdehyde(MDA)and GSH/GSSG were detected by microplate reader.The expression levels of reactive oxygen species(ROS),Fe2+and lipid peroxide(LPO)in B16F10 cells were detected by flow cytometry.Results Compared with melanoma cells transferred to lymph nodes,the non-metastatic group had a higher degree of ferroptosis and a lower expression level of SLC7A11.The cytotoxicity of MB and MB-PDT on B16F10 was dose-dependent(P＜0.000 1).The results of scratch assay showed that MB-PDT inhibited the migration ability of B16F10 cells(P＜0.001).Compared with the control group,after MB-PDT treatment,the expression levels of GPX4,SLC7A11 and GSH/GSSG were decreased(P＜0.05),and the expression levels of ROS,Fe2+,LPO and MDA were increased(P＜0.05)in B16F10 cells.Pretreatment with Ferrostatin-1,an inhibitor of ferroptosis,inhibited MB-PDT-induced cytotoxicity and ferroptosis in B16F10 cells(P＜0.05).Conclusion MB-PDT can induce ferroptosis of melanoma cells by inhibiting SLC7A11 expression,thereby inhibiting the progression of melanoma.

Objective To investigate the relationship between the expression of programmed death ligand 1(PD-L1)and hypoxia inducible factor-1α(HIF-1 α)with the clinical pathological characteristics and prognosis of gastric cancer patients.Methods The cancer tissues of 100 gastric cancer patients who underwent radical gastrectomy at the First Hospital of Handan City from July 2019 to July 2020 were selected as the research subjects,and their adjacent tissues(normal tissues ≥ 5 cm from the cancer tissues)were as the control group.Immunohistochemical detection of PD-L1 and HIF-1 α was performed by SP method.Spearman correlation analysis was used to analyze PD-L1 and HIF-1 α in gastric cancer tissues.Kaplan-Meier method was used to analyze the 3-year survival relationship of gastric cancer patients.The influencing factors of prognosis and death in patients with gastric cancer were analyzed by univariate and multivariate Cox regression.Results Among 100 gastric cancer patients,52 were PD-L1 positive and 48 were negative;67 were HIF-1 α positive and 33 were HIF-1 α negative,the positive expression rates of PD-L1 and HIF-1α in gastric cancer tissues were 52.00％and 67.00％,respectively,which were obviously higher than those in adjacent tissues(11.00％、18.00％),the difference was statistically significant(P＜0.05).Spearman correlation analysis showed that the expression of PD-L1 was positively correlated with that of HIF-1α in gastric cancer tissues(r=0.730,P＜0.001).The expressions of PD-L1 and HIF-1α in patients with gastric cancer were correlated with TNM stage,lymph node metastasis and local invasion(P＜0.05).The 3-year overall survival rate of gastric cancer patients was 48.00％after surgery,and the 3-years survival rate of patients with positive expression of PD-L1 and HIF-1α were 28.85％and 31.34％,which were lower than those of patients with negative expression of PD-L1 and HIF-1α(68.75％and 81.82％)(Log rank x2=25.155,P＜0.001.Log rank x2=24.552,P＜0.001).Moreover,positive expression of PD-L1 and HIF-1α,TNM staging of Ⅲ-Ⅳ,lymph node metastasis,and local infiltration were independent risk factors for prognosis and death in gastric cancer patients(P＜0.05).Conclusion Both PD-L1 and HIF-1α are highly expressed in cancer tissues of gastric cancer patients,and they are positively correlated.They are also associated with clinical pathological features such as TNM staging,lymph node metastasis,and poor prognosis.

Objective To investigate the promoting effect of quercetin(QR)on diabetic wound repair and its possible mechanism.Methods HUVEC cell injury model was induced by 33.36 mmol/L high glucose.The cells were divided into normal control group(NG group),mannitol group(MA group),high glucose group(HG group),low dose quercetin+high glucose group(QR 5 μmol/L+HG group),high dose quercetin+high glucose group(QR 20 μmol/L+HG group).Treatment for 24 hours;CCK-8 and Edu were used to measure cell viability.Cell scratch test and Transwell migration assay were used to detect cell migration ability.Matrigel tube formation assay was used to measure the angiogenesis ability.The levels of IL-1β,IL-18 and TNF-α were detected by ELISA.Network pharmacology was used to screen the potential targets of QR on diabetic wounds.Western blot was used to detect the expression of NLRP3 inflammasome,PI3K/Akt/GSK-3β pathway and its phosphorylation.The diabetic wound model of C57 mice was established and divided into STZ group and QR+STZ group.The wound healing rate within 14 days was calculated and analyzed,and the wound tissue was stained with HE and Masson on the 14th day.Results CCK-8 and Edu results showed that a high dose of QR could improve the viability of huvecs induced by high glucose.Scratch test and Transwell migration test showed that high dose of QR could enhance the migration ability of HUVEC cells induced by high glucose.Matrigel tube formation assay showed that high dose of QR could enhance the angiogenesis ability of HUVEC cells induced by high glucose.ELISA results showed that high dose of QR could inhibit the secretion of IL-1β,IL-18 and TNF-α induced by high glucose.Network pharmacology screening results showed that PI3K/Akt/GSK-3β pathway was significantly enriched.Western blot showed that 20 μmol/L QR could increase the phosphorylation level of PI3K/Akt/GSK-3β pathway and inhibit the expression of NLRP3 inflammasome,which could be reversed by the addition of PI3K phosphorylation inhibitor LY294002.Animal experiments have shown that QR can promote diabetic wound healing by enhancing granulation tissue growth,epithelial regeneration,and collagen deposition.Conclusion QR can improve HUVEC injury induced by high glucose and promote wound healing in diabetic mice,and its mechanism may be related to inhibiting NLRP3 expression through PI3K/Akt/GSK-3β pathway.

Objective To investigate the promoting effect of quercetin(QR)on diabetic wound repair and its possible mechanism.Methods HUVEC cell injury model was induced by 33.36 mmol/L high glucose.The cells were divided into normal control group(NG group),mannitol group(MA group),high glucose group(HG group),low dose quercetin+high glucose group(QR 5 μmol/L+HG group),high dose quercetin+high glucose group(QR 20 μmol/L+HG group).Treatment for 24 hours;CCK-8 and Edu were used to measure cell viability.Cell scratch test and Transwell migration assay were used to detect cell migration ability.Matrigel tube formation assay was used to measure the angiogenesis ability.The levels of IL-1β,IL-18 and TNF-α were detected by ELISA.Network pharmacology was used to screen the potential targets of QR on diabetic wounds.Western blot was used to detect the expression of NLRP3 inflammasome,PI3K/Akt/GSK-3β pathway and its phosphorylation.The diabetic wound model of C57 mice was established and divided into STZ group and QR+STZ group.The wound healing rate within 14 days was calculated and analyzed,and the wound tissue was stained with HE and Masson on the 14th day.Results CCK-8 and Edu results showed that a high dose of QR could improve the viability of huvecs induced by high glucose.Scratch test and Transwell migration test showed that high dose of QR could enhance the migration ability of HUVEC cells induced by high glucose.Matrigel tube formation assay showed that high dose of QR could enhance the angiogenesis ability of HUVEC cells induced by high glucose.ELISA results showed that high dose of QR could inhibit the secretion of IL-1β,IL-18 and TNF-α induced by high glucose.Network pharmacology screening results showed that PI3K/Akt/GSK-3β pathway was significantly enriched.Western blot showed that 20 μmol/L QR could increase the phosphorylation level of PI3K/Akt/GSK-3β pathway and inhibit the expression of NLRP3 inflammasome,which could be reversed by the addition of PI3K phosphorylation inhibitor LY294002.Animal experiments have shown that QR can promote diabetic wound healing by enhancing granulation tissue growth,epithelial regeneration,and collagen deposition.Conclusion QR can improve HUVEC injury induced by high glucose and promote wound healing in diabetic mice,and its mechanism may be related to inhibiting NLRP3 expression through PI3K/Akt/GSK-3β pathway.

Objective:To explore the training methods to improve the cosmetic suture technique of junior surgeons.Methods:The training course of cosmetic suture techniques was carried out for junior residents such as plastic surgery professional graduate students and standardized training residents, through the study of the basic knowledge of cosmetic suture and the practical training of minimally invasive principles, to help them master cosmetic suture techniques systematically for the need of future clinical practice. Compared with the level of operation skills before and after cosmetic suture training in different groups (graduate students of plastic surgery and other majors, standardized training residents of plastic surgery and other majors), the residents' performance had been significantly improved. The incisions healing after half a year follow-up in cases they sutured were observed.Results:After cosmetic suture training, the suture skills of the students were significantly improved, and there was a significant correlation with the time of clinical practice. The incidence of incisions inflammation and postoperative scar formation in the cases sutured by the students with practice for 6 months was significantly lower than those for 1 month (6.42% vs 21.67%, 1.13% vs 5.56% respectively).Conclusions:The training of cosmetic suture techniques is very important to improve the professional ability of junior residents in plastic and cosmetic surgery, and it needs a long time practice to master the suture skills.

This study introduces that we have established a clinical case database of scar prevention and treatment with medical histories, diagnosis and treatment ideas, and case expansion according to the scar diseases that professional postgraduates of plastic surgery need to master, and applied it to the practical teaching for plastic surgery postgraduates. Through the comprehensive evaluation from three aspects of mastering basic knowledge, applying knowledge and developing discipline, it's suggested that the professional postgraduates can better grasp the knowledge of scar prevention and treatments after learning from the clinical scar database, and the application of the teaching case database has further improved students' teaching and learning participations and innovative thinking abilities.

Objective:To investigate the clinical effect of botulinum toxin type A injection in the prevention and treatment of frontal traumatic scars immediately after operation.Methods:From January 2016 to January 2018, among the 152 patients with frontal trauma admitted to the Department of Plastic Surgery, Daping Hospital, Army Military Medical University, patients with wounds almost perpendicular to the Langer line were selected. Immediately after debridement and suture, botulinum toxin type A was injected with a concentration of 2 U botulinum toxin per 0.10 ml of solution, 0.05 to 0.10 ml (1 to 2 U) per 1 cm was injected into the frontal muscle according to frontal muscle strength.With reference to the observer scar assessment scale and Vancouver scar scale, the color, width, texture, thickness, pain, and itching of the postoperative scar of the frontal wound were evaluated at follow-up.Results:A total of 44 patients were included, 22 males and 22 females, aged (31.5 ± 8.4) years old, with a wound length of 1-12 cm(average, 3.5 cm) and a Botox injection volume around wounds of 0.10-0.90 ml (2-18 U) . The patients were followed up for 6 to 24 months. One patient had mild to moderate ptosis, which completely disappeared within one month. Within 6 weeks after operation, the scars in 44 patients were pink and hard. 3-6 months later, the scars gradually became soft, narrow and flat with similiar skin color, no pain and itching left in 42 patients. The satisfaction rate was 95.5% (42/44). Two patients were dissatisfied because of scar pigmentation and width.Conclusions:Immediate injection of botulinum toxin type A after debridement and suture of the frontal trauma wound can effectively prevent and improve scar appearance with a high rate of satisfaction.

Objective:To investigate the clinical effect of botulinum toxin type A injection in the prevention and treatment of frontal traumatic scars immediately after operation.Methods:From January 2016 to January 2018, among the 152 patients with frontal trauma admitted to the Department of Plastic Surgery, Daping Hospital, Army Military Medical University, patients with wounds almost perpendicular to the Langer line were selected. Immediately after debridement and suture, botulinum toxin type A was injected with a concentration of 2 U botulinum toxin per 0.10 ml of solution, 0.05 to 0.10 ml (1 to 2 U) per 1 cm was injected into the frontal muscle according to frontal muscle strength.With reference to the observer scar assessment scale and Vancouver scar scale, the color, width, texture, thickness, pain, and itching of the postoperative scar of the frontal wound were evaluated at follow-up.Results:A total of 44 patients were included, 22 males and 22 females, aged (31.5 ± 8.4) years old, with a wound length of 1-12 cm(average, 3.5 cm) and a Botox injection volume around wounds of 0.10-0.90 ml (2-18 U) . The patients were followed up for 6 to 24 months. One patient had mild to moderate ptosis, which completely disappeared within one month. Within 6 weeks after operation, the scars in 44 patients were pink and hard. 3-6 months later, the scars gradually became soft, narrow and flat with similiar skin color, no pain and itching left in 42 patients. The satisfaction rate was 95.5% (42/44). Two patients were dissatisfied because of scar pigmentation and width.Conclusions:Immediate injection of botulinum toxin type A after debridement and suture of the frontal trauma wound can effectively prevent and improve scar appearance with a high rate of satisfaction.

OBJECTIVE:To prepare and characterize Ibuprofen (IBU) nano-powder,and to investigate its transdermal ability in vitro. METHODS:Using chloroform-ethanol(7:3,V/V)as organic phase,deionized water as aqueous phase and polysorbate 80 as surfactant,the emulsification method was used to prepare IBU nano-powder. Laser granulometric analysis,Fourier transform in-frared spectroscopy(FT-IR),X-ray diffraction(XRD),differential scanning calorimetry(DSC)were used to characterize IBU na-no-powder. IBU nano-powder was compared with bulk drug in respects of saturation solubility,dissolution rate and transdermal rate in vitro. RESULTS:The optimum condition was as follows that the concentration of polysorbate 80 was 5 mg/mL;the volume ra-tio of water phase-organic phase was 40:1;the concentration of IBU was 250 mg/mL;homogenate speed was 5000 r/min;homog-enate time was 2 min. Prepared IBU nano-powder was polyporous crumbly coralliform,and its chemical structure kept stable;the nano-powder changed from crystal to amorphous state;the particle size was 179.6 nm,and drug-loading amount was 8.99%;satu-ration solubility,dissolution rate and transdermal rate of IBU nano-powder were 148,1.23 and 4.08 times of bulk drug. CONCLU-SIONS:The prepared IBU nano-powder shows good water-solubility and percutaneous permeability.