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 clarify the effect of methotrexate on blood uric acid levels and the incidence of hyperuricemia in patients with rheumatic and musculoskeletal diseases (RMDs).Methods:The clinical data were collected from 349 patients with RMDs who took methotrexate for more than 52 weeks and 429 patients with RMDs who did not take methotrexate, who were treated at Anqing Medical Center of Auhui Medical University from June 1, 2022 to June 30, 2024, to compare the differences in serum uric acid concentration and the incidence of hyperuricemia before and after 24 weeks of methotrexate administration in the two groups of patients with RMDs. The changes in serum uric acid concentration and serum creatinine value in the MTX na?ve patients who had taking MTX for 0, 24 and 52 weeks were compared. The relationship between serum uric acid concentration and methotrexate dosage was analyzed. Measurement data were compared using t-test or ANOVA, repeated measures analysis of variance, and count data were compared using χ2 test. Results:①At week 0, there was no significant difference in serum uric acid concentration [(300±63)μmol/L vs. (306±64)μmol/L, t=-1.416, P=0.157] and the incidence of hyperuricemia [9.3%(40/429) vs. 10.3%(36/349) , χ2=0.215, P=0.643] between the two groups. At week24, the serum uric acid concentration (307±70)μmol/L vs. (246±89)μmol/L was statistically significantly ( t=10.909, P<0.001) different. The incidence of hyperuricemia (11.0%, 47/429) vs. (4.6%, 16/349), was statistically significantly different ( χ2=10.497, P<0.001). There was a statistically significant difference in serum uric acid concentration between week 0 and week 24 in the methotrexate group ( t=10.237, P<0.001), and there was a statistically significant difference in the incidence of hyperuricemia ( χ2=8.312, P=0.004). ②The overall serum uric acid concentrations at week 0, weeks 24, and weeks 52 were (306±64)μmol/L, (246±89)μmol/L, and (247±66)μmol/L, respectively. The difference in overall serum uric acid concentration was statistically significant ( F= 29.506, P<0.001). There was no significant difference in serum uric acid concentration between weeks 24 and 52 ( P=1.000). There were significant differences in serum creatinine levels between weeks 0, 24 and 52 ( P<0.001). There was no significant difference in serum creatinine levels between weeks 0 ,52, weeks 24 and 52 ( P=0.077, P=1.000). There were statistically significant differences in the overall serum uric acid concentration and serum creatinine value at weeks 0, 24 and 52 of medication ( P<0.001).③ There was no significant difference in serum uric acid concentration before and after taking hydroxychloroquine, cyclosporine, tripterygium wilfordii, mycophenolate mofetil, tofacitinib, etanercept and adalimumab alone for weeks 0 and 24(all P>0.05). ④There was no significant difference in serum uric acid concentration between patients taking different doses of methotrexate (7.5 mg once weekly, 10 mg once weekly, 12.5 mg once weekly, 15 mg once weekly) at weeks 0 and 24 weeks(all P>0.05). Conclusion:MTX, as an anti-rheumatic drug, reduces the serum uric acid level and the incidence of hyperuricemia in patients with RMDs during the treatment.

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 clarify the effect of methotrexate on blood uric acid levels and the incidence of hyperuricemia in patients with rheumatic and musculoskeletal diseases (RMDs).Methods:The clinical data were collected from 349 patients with RMDs who took methotrexate for more than 52 weeks and 429 patients with RMDs who did not take methotrexate, who were treated at Anqing Medical Center of Auhui Medical University from June 1, 2022 to June 30, 2024, to compare the differences in serum uric acid concentration and the incidence of hyperuricemia before and after 24 weeks of methotrexate administration in the two groups of patients with RMDs. The changes in serum uric acid concentration and serum creatinine value in the MTX na?ve patients who had taking MTX for 0, 24 and 52 weeks were compared. The relationship between serum uric acid concentration and methotrexate dosage was analyzed. Measurement data were compared using t-test or ANOVA, repeated measures analysis of variance, and count data were compared using χ2 test. Results:①At week 0, there was no significant difference in serum uric acid concentration [(300±63)μmol/L vs. (306±64)μmol/L, t=-1.416, P=0.157] and the incidence of hyperuricemia [9.3%(40/429) vs. 10.3%(36/349) , χ2=0.215, P=0.643] between the two groups. At week24, the serum uric acid concentration (307±70)μmol/L vs. (246±89)μmol/L was statistically significantly ( t=10.909, P<0.001) different. The incidence of hyperuricemia (11.0%, 47/429) vs. (4.6%, 16/349), was statistically significantly different ( χ2=10.497, P<0.001). There was a statistically significant difference in serum uric acid concentration between week 0 and week 24 in the methotrexate group ( t=10.237, P<0.001), and there was a statistically significant difference in the incidence of hyperuricemia ( χ2=8.312, P=0.004). ②The overall serum uric acid concentrations at week 0, weeks 24, and weeks 52 were (306±64)μmol/L, (246±89)μmol/L, and (247±66)μmol/L, respectively. The difference in overall serum uric acid concentration was statistically significant ( F= 29.506, P<0.001). There was no significant difference in serum uric acid concentration between weeks 24 and 52 ( P=1.000). There were significant differences in serum creatinine levels between weeks 0, 24 and 52 ( P<0.001). There was no significant difference in serum creatinine levels between weeks 0 ,52, weeks 24 and 52 ( P=0.077, P=1.000). There were statistically significant differences in the overall serum uric acid concentration and serum creatinine value at weeks 0, 24 and 52 of medication ( P<0.001).③ There was no significant difference in serum uric acid concentration before and after taking hydroxychloroquine, cyclosporine, tripterygium wilfordii, mycophenolate mofetil, tofacitinib, etanercept and adalimumab alone for weeks 0 and 24(all P>0.05). ④There was no significant difference in serum uric acid concentration between patients taking different doses of methotrexate (7.5 mg once weekly, 10 mg once weekly, 12.5 mg once weekly, 15 mg once weekly) at weeks 0 and 24 weeks(all P>0.05). Conclusion:MTX, as an anti-rheumatic drug, reduces the serum uric acid level and the incidence of hyperuricemia in patients with RMDs during the treatment.

Objective To explore the protective role of DNA damage induced transcription factor 4(DDIT4)in oxidative stress injury in dermal papilla cells and its underlying mechanisms.Methods Dermal papilla cells were exposed to UVA and H2O2 to establish cellular model of oxidative stress.CCK-8 assay was used to detect cell viability under different treatment conditions,and the production of intracellular reactive oxygen species(ROS)was detected using 2',7'-dichlorofluorescein diacetate(DCFH-DA).Autophagic vesicles were observed with electron microscopy.Western blotting was employed to measure the expression of DDIT4 and autophagy-related molecules,including microtubule-associated protein 1 light chain 3(LC3),ubiquitin-binding protein(P62),mammalian target of rapamycin(mTOR),and p-mTOR.Results UVA and H2O2 resulted in more production of ROS(P＜0.05)and decreased viability of dermal papilla cells(P＜0.05).DDIT4 expression was increased in dermal papilla cells under oxidative stress(P＜0.05),and the antioxidant N-acetyl-L-cysteine(NAC)could effectively inhibit this effect(P＜0.05).After treatment with UVA or H2O2,cell autophagy was enhanced in dermal papilla cells,characterized by an increase in the number of autophagosomes and an increased LC3 Ⅱ/Ⅰ ratio(P＜0.05),a decrease in P62 expression(P＜0.05),and 3-methyladenine(3-MA)blocking autophagy led to further reduced cell viability(P＜0.05)and increased intracellular ROS production(P＜0.05).Conversely,rapamycin(RAPA)increased autophagy level and improved the viability of dermal papilla cells under oxidative conditions(P＜0.05),and reduced the generation of intracellular ROS(P＜0.05).Additionally,down-regulation of DDIT4 weakened autophagy in dermal papilla cells under oxidative stress,reduced LC3 Ⅱ/Ⅰ(P＜0.05),increased p-mTOR/mTOR and P62(P＜0.05),inhibited cell viability(P＜0.05),and enhanced intracellular ROS production(P＜0.05).Conclusion DDIT4 may alleviate oxidative stress injury in dermal papilla cells through autophagy.

Objective:To analyze the operational efficiency and Total Factor Productivity (TFP) of township health centers in Ningxia from 2013 to 2022,in order to provide decision-making basis for optimize the resource allocation of township health centers in Ningxia. Methods:The BCC model of Data Envelopment Analysis (DEA) and Malmquist productivity index were used to analyze the operation efficiency and TFP of township health centers of Ningxia. Results:(1) From 2013 to 2022,the personnel input in Ningxia township health centers increased by 45.80％,the number of outpatient visits had no significant increase,and the number of inpatient visits decreased by 45.40％. (2) The average comprehensive technical efficiency of resource allocation was 0.658,the average of pure technical efficiency and scale efficiency were 0.747 and 0.880. (3) The average value of TFP index was greater than 1 in 7 counties,and less than 1 in the other 15 counties. Conclusion:The efficiency of township health centers in Ningxia was low. It is of great significance to improve the resource allocation and management ability of township health centers and strengthen the policy support for improving the operation efficiency of township health centers.

Objective:To analyze the operational efficiency and Total Factor Productivity (TFP) of township health centers in Ningxia from 2013 to 2022,in order to provide decision-making basis for optimize the resource allocation of township health centers in Ningxia. Methods:The BCC model of Data Envelopment Analysis (DEA) and Malmquist productivity index were used to analyze the operation efficiency and TFP of township health centers of Ningxia. Results:(1) From 2013 to 2022,the personnel input in Ningxia township health centers increased by 45.80％,the number of outpatient visits had no significant increase,and the number of inpatient visits decreased by 45.40％. (2) The average comprehensive technical efficiency of resource allocation was 0.658,the average of pure technical efficiency and scale efficiency were 0.747 and 0.880. (3) The average value of TFP index was greater than 1 in 7 counties,and less than 1 in the other 15 counties. Conclusion:The efficiency of township health centers in Ningxia was low. It is of great significance to improve the resource allocation and management ability of township health centers and strengthen the policy support for improving the operation efficiency of township health centers.

OBJECTIVE：To ex plore the protective effects of Longbie capsule contained serum （called“LBJN”for short ）on the apoptosis of chondrocytes induced by YAP inhibitor verteporfin and its mechanism. METHODS ：Primary human knee osteoarthritis（OA）chondrocytes were extracted by two-step enzymatic digestion ，and then identif ied by toluidine blue staining and type Ⅱ collagen immunofluorescence staining. The effects of 2，5 μmol/L verteporfin alone or combined with 5％LBJN on cell apoptosis were detected by flow cytometry. Solvent control （0.1％ DMSO）and 5％ LBJN were set. Western blot assay was adopted to detect the expression of apoptosis related proteins （YAP，Bcl-2，cleaved-caspase-3） after treated with 0.1％DMSO（solvent control ），2 μmol/L verteporfin，2 μmol/L verteporfin+5％LBJN 和 0（blank control ），2.5％ LBJN and 5％ LBJN for 48 h. The expression of autophagy related proteins （mTOR，Beclin-1，LC3A/B） after treated with 0 （blank control ），2.5％，5％ LBJN for 48 h were det ected by Western blot assay. RESULTS ：The isolated cells accorded with the characteristics of chondrocytes. Compared with 0.1％DMSO， the apoptosis rates of cells were increased significantly after treated with 2，5 μmol/L verteporfin（P＜0.05），and the effects of the two concentrations were similar （P＞0.05）. Compared with verteporfin alone ，2，5 μmol/L verteporfin combined with 5％LBJN could significantly decrease the apoptotic rate of cells （P＜0.05）. Compared with 0.1％DMSO，the protein expression of YAP and Bcl-2 were decreased significantly after treated with 2 μ mol/L verteporfin （P＜0.05）， while the protein expression of cleaved-caspase-3 were increased significantly （P＜0.05）. Compared with 2 μmol/L verteporfin，protein expression of YAP and Bcl-2 were increased significantly after treated with 2 μmol/L verteporfin+5％LBJN（P＜0.05），while the protein expression of cleaved-caspase-3 were decreased significantly （P＜0.05）. Compared with blank control ，the protein expression of YAP ，Bcl-2 and Beclin-1 were increased significantly after treated with 2.5％，5％LBJN（P＜0.05），while protein expression of cleaved-caspase- 3 and mTOR were decreased significantly （P＜0.05）. CONCLUSIONS ：LBJN can block the apoptosis of chondrocytes induced by YAP inhibitor verteporfin ，and its mechanism may be related to regulating the expression of apoptosis related proteins and enhancing autophagy of chondrocytes.

Objective:To explore the efficacy of doctor-nurse co-led care involving education and engagement of patients on improving compliance of patients, and a treat-to-target urate-lowering rate for gout.Methods:Nurses were trained in practice management of gout. Patients diagnosed with gout in the departments of rheumatology and immunology of Anqing Municipal Hospital in Anhui Province were prospectively enrolled from January 1 to June 30, 2019. The patients were divided into the continuous-care group and the conventional management group by random number table method. The patients of continuous-care group received telephone follow-up, WeChat tracking and regular face-to-face communication. The patients of conventional management group were told to follow up regularly in the outpatient department, and the nurses did not follow up. Patients were evaluated before intervention and 12 months after intervention. The treat-to-target rate of blood uric acid and the frequency of gout flares were observed.Results:After 12 months of intervention, the patients of serum uric acid concentrations below 360 μmol/L were 92.39% (85/92) in the continuous-care group, and 26.74% (23/86) in the conventional management group. There was significant difference ( χ2 value was 80.282, P<0.001). After 12 months of intervention, the average serum uric acid concentration of patients in the continuous-care group was (301.6±61.4) μmol/L, and that in the conventional management group was (419.0±98.0) μmol/L, both of which were significantly lower than before intervention, continuous-care group (466.1±119.7) μmol/L, conventional management group (477.8±113.1) μmol/L. But the average serum uric acid concentration of patients in the continuous-care group was significantly lower than that in the conventional management group. There was significant difference between them ( t value was 96.678, P<0.001). At the end of 12 months, the patients of uric-acid-lowering therapy increased in both groups. The proportion of patients was 94.56% (87/92) in the continuous-care group, which was significantly higher than that in the conventional management group (58.14%, 50/86), there was significant difference ( χ2 value was 33.260, P<0.001). Conclusions:The mode of continuing nursing combined with specialized physician-led treatment can significantly improve the compliance and the control rate of treat-to-target for gout, and this management method is simple and feasible which provides a new management concept for clinical treatment of gout.

Objective: To investigate the prevalence of metabolic disorder among the high school students in Guangzhou. Methods: A cross-sectional study among 4 620 high school students selected with stratified cluster sampling was conducted in Guangzhou. Height, weight, blood pressure, fasting plasma glucose (FPG) and serum lipid of TC, TG were measured. Household socioeconomic status, medical history, physical activities as well as personal diet and study habits were acquired with structured questionnaire. Results: The prevalence of central obesity was 11.5%. The overall prevalence of IFG and DM was 6.9%. The overall prevalence of both hypercholesteremia and hypertriglyceridemia was 13.4%. The prevalence of hypertension was 11.6%. With multivariable logistic regression, the results showed that: fast eating was associated with increased risk of central obesity (OR=1.79, 95%CI=1.47-2.18) and hypercholesteremia/hypertriglyceridemia (OR=1.27，95%CI=1.04-1.54). While students who ate slow had low risk of central obesity (OR=0.47, 95%CI=0.33-0.68), hypercholesteremia/hypertriglyceridemia(OR=0.71, 95%CI=0.54-0.94) and hypertension (OR=0.65, 95%CI=0.49-0.88). Students with sugar-sweetened beverages once a day or more had higher risk of central obesity (OR=1.60, 95%CI=1.08-2.38) while students without sugar-sweetened beverages consumption had low risk of IFG/DM (OR=0.57, 95%CI= 0.43-0.77) than those who consumed 1-2 times/week. Students without eating out habit had lower risk of hypercholesteremia /hypertriglyceridemia than those with eating out for 1-2 times/week (OR=0.78, 95%CI=0.62-0.98), and students who had lunch out of home or canteen had higher risk of hypercholesteremia /hypertriglyceridemia than those who ate at canteen (OR=1.38，95%CI=1.06-1.81). Conclusion The prevalence of metabolic disorder among high school students in Guangzhou is at a relatively high level. High eating speed, heavy sugared beverages intake and frequently eating out habits are associated with risk of metabolic disorder.