Objective:To develop a self-locked DNAzyme nanoprobe-based fluorescence amplification strategy for wash-free and ultrasensitive detection of breast cancer-derived small extracellular vesicles (sEV).Method:A DNAzyme self-locked probe was designed to recognize the epithelial cell adhesion molecule (EpCAM) specifically expressed on breast cancer-derived sEVs. Upon binding to EpCAM, the DNAzyme-lock structure was opened, restoring the DNAzyme cleavage activity. The activated DNAzyme then cyclically cleaved the RNA site on the substrate strand. Fluorescently labeled substrate strands were used to detect sEVs at varying concentrations, and the detection limit and linear range were determined.Results:The DNAzyme self-locked probe successfully identified breast cancer-derived sEVs and generated a fluorescent signal through cyclic cleavage. The proposed method achieved wash-free detection of sEVs, with the fluorescence intensity showing a strong linear correlation with sEV concentration ( R2=0.98). The linear detection range was 1.0×10 2-1.0×1.0 7 particles/μl, with a detection limit of 59 particles/μl. Conclusion:This study established a wash-free and highly sensitive strategy for quantifying breast cancer-derived sEVs, which provides a promising technical approach for the early diagnosis of cancer.

Objective To investigate necroptosis-related diagnostic biomarkers of bronchopulmo-nary dysplasia(BPD)and their relationships with the immune microenvironment through the analysis of necroptosis-related genes(NRGs)in BPD.Methods The dataset GSE32472 was downloaded from the Gene Expression Omnibus(GEO)database,and NRGs were obtained from the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway and Gene Cards databases.Differentially expressed necroptosis-related genes(DE-NRGs)were screened,and their biological functions and pathways were explored through functional enrichment analysis.Machine learning algorithms,inclu-ding least absolute shrinkage and selection operator(LASSO)and support vector machine-recursive feature elimination(SVM-RFE),were applied to screen feature genes.The Cell-type Ⅰdentification By Estimating Relative Subsets of RNA Transcripts(CIBERSORT)algorithm and the Estimation of Stromal and Immune Cells in Malignant Tumor Tissues using Expression Data(ESTIMATE)algo-rithm were used to explore the immune infiltration characteristics of BPD.Spearman correlation anal-ysis between feature genes and immune cells was performed using the"corrplot"package in R lan-guage.Results A total of 19 DE-NRGs were identified.The main biological functions and path-ways of DE-NRGs included the regulation of necroptosis and inflammatory responses.Three feature genes,namely flotillin-2(FLOT2),CASP8 and FADD-like apoptosis regulators(CFLAR),and charged multivesicular body protein 7(CHMP7),were further screened to construct a nomogram.In the validation sets GSE8586 and GSE188944,the area under the curve(AUC)values were all greater than 0.7.CIBERSORT analysis revealed that BPD group presented a higher proportion of naive B cells,neutrophils,eosinophils and resting mast cells compared to control group(P＜0.05).Meanwhile,the proportion of CD8+T cells,CD4+naive T cells,CD4+resting memory T cells,regulatory T cells,resting natural killer(NK)cells,M0 macrophages,M2 macrophages and activated dendritic cells was lower than that in the control group(P＜0.05).ESTIMATE analysis showed that the stromal score in the BPD group was higher than that in the control group,while the immune score was lower,with statistically significant differences(P＜0.05).Correlation analysis between the three feature genes and ESTIMATE scores indicated that FLOT2 and CFLAR were posi-tively correlated with the stromal score and negatively correlated with the immune score,whereas CHMP7 was positively correlated with the immune score and negatively correlated with the stromal score.Conclusion The three necroptosis-related feature genes can serve as diagnostic biomarkers for BPD-related necroptosis,with high diagnostic efficacy.They may play an important roles through immune mechanisms,providing new insights and theoretical references for the early diagnosis and immune intervention of BPD.

Objective To explore causal relationship between 25-hydroxyvitamin D[25(OH)D]and ankylosing spondylitis(AS).Methods Genetic data of 25(OH)D and AS were extracted from the genome-wide association study.The causal effect of 25(OH)D on AS was estimated by MR-Egger regression method,weighted median,inverse variance weighted(IVW),simple mode and weighted mode,and sensitivity analysis was conducted for verification.Results The IVW results indicated that there was a causal relationship between 25(OH)D concentration and AS(OR=0.805,95％CI:0.686-0.944,P=0.008),and the maximum likelihood ratio(OR=0.799,95％CI:0.678-0.940,P=0.007)showed consistent results.The IVW results of the reverse Mendelian randomization study showed that there was no causal relationship between the two(OR=1.019,95％CI:0.995-1.043,P=0.110).In addition,MR-Egger intercept,Cochran Q test,"leave-one-out"and MR-PRESSO analysis showed no horizontal pleipotency or heterogeneity.Conclusion There may be a genetic causal relationship between the concentration of 25(OH)D and the onset of AS.AS cannot cause changes in the concentration of 25(OH)D in the body.

Objective To explore causal relationship between 25-hydroxyvitamin D[25(OH)D]and ankylosing spondylitis(AS).Methods Genetic data of 25(OH)D and AS were extracted from the genome-wide association study.The causal effect of 25(OH)D on AS was estimated by MR-Egger regression method,weighted median,inverse variance weighted(IVW),simple mode and weighted mode,and sensitivity analysis was conducted for verification.Results The IVW results indicated that there was a causal relationship between 25(OH)D concentration and AS(OR=0.805,95％CI:0.686-0.944,P=0.008),and the maximum likelihood ratio(OR=0.799,95％CI:0.678-0.940,P=0.007)showed consistent results.The IVW results of the reverse Mendelian randomization study showed that there was no causal relationship between the two(OR=1.019,95％CI:0.995-1.043,P=0.110).In addition,MR-Egger intercept,Cochran Q test,"leave-one-out"and MR-PRESSO analysis showed no horizontal pleipotency or heterogeneity.Conclusion There may be a genetic causal relationship between the concentration of 25(OH)D and the onset of AS.AS cannot cause changes in the concentration of 25(OH)D in the body.

Objective:To develop a self-locked DNAzyme nanoprobe-based fluorescence amplification strategy for wash-free and ultrasensitive detection of breast cancer-derived small extracellular vesicles (sEV).Method:A DNAzyme self-locked probe was designed to recognize the epithelial cell adhesion molecule (EpCAM) specifically expressed on breast cancer-derived sEVs. Upon binding to EpCAM, the DNAzyme-lock structure was opened, restoring the DNAzyme cleavage activity. The activated DNAzyme then cyclically cleaved the RNA site on the substrate strand. Fluorescently labeled substrate strands were used to detect sEVs at varying concentrations, and the detection limit and linear range were determined.Results:The DNAzyme self-locked probe successfully identified breast cancer-derived sEVs and generated a fluorescent signal through cyclic cleavage. The proposed method achieved wash-free detection of sEVs, with the fluorescence intensity showing a strong linear correlation with sEV concentration ( R2=0.98). The linear detection range was 1.0×10 2-1.0×1.0 7 particles/μl, with a detection limit of 59 particles/μl. Conclusion:This study established a wash-free and highly sensitive strategy for quantifying breast cancer-derived sEVs, which provides a promising technical approach for the early diagnosis of cancer.

Objective:To explore the clinical effects of anterograde sural neurovascular flap in repairing skin and soft tissue defect around the knee.Methods:Nine patients with skin and soft tissue defect around the knee admitted to Beijing Fengtai YouAnMen Hospital from May 2011 to December 2018, were included in this retrospective descriptive study, including 8 males and 1 female, aged 16 to 65 years. The wound area after debridement ranged from 8 cm×5 cm to 18 cm×10 cm. Anterograde sural neurovascular flap was used to repair the wounds in 9 patients, with the area ranging from 9 cm×6 cm to 20 cm×12 cm. The donor sits of flaps in 2 patients were closed and sutured directly, and the donor sits of flaps in 7 patients were repaired with medial split-thickness skin graft of the ipsilateral thigh. The flap survival, complications, and follow-up after operation were recorded.Results:The flaps survived and the blood supply was good in 8 patients and the wounds were closed. One patient developed skin ischemic necrosis which was cured after three weeks of dressing change. All the skin grafts in the donor site of flap in 7 patients survived. In 6 months to 5 years of follow-up after surgery, the skin flap had good texture, color, and shape, and normal sensation. Except for one patient whose knee had poor recovery of function, the knee joint function of the other patients recovered well.Conclusions:The anterograde sural neurovascular flap has the advantages of high survival rate, satisfactory appearance and functional recovery post surgery, and is an ideal flap for repairing the skin and soft tissue defect around the knee.

Objective:To explore the characteristics of illness perceptions and its effect on quality of life in adult epileptics.Methods:A cross-sectional study was conducted to evaluate 98 adult patients with epilepsy and 75 diabetic patients hospitalized at the same period.The Chinese illness perception questionnaire revised edition(CIPQ-R) was used to evaluate the differences in illness perception factors between the two groups.The quality of life in epilepsy-31(QOLIE-31) was used to assess the quality of life of 98 adult epilepsy patients and 55 healthy people, and the correlation analysis and multivariate regression analysis were used to investigate the effect of illness perceptions on the quality of life of epileptic patients.Results:The emotional representations factor score of CIPQ-R in the epilepsy group(21.73±5.79) was higher than that in the diabetes group (18.76±5.42) ( P<0.01). The scores of timeline acute/chronic, personal control and illness coherence factors of CIPQ-R in the epilepsy group((16.47±4.68), (20.91±4.03), (15.21±4.44)) were lower than those in the diabetes group ((19.16±4.75), (22.68±3.90), (16.56±3.73)) ( P<0.05). The scores of multiple factors in the quality of life scale of patients with epilepsy were lower than those in healthy controls( t=-2.264--8.203, P<0.01). Correlation analysis showed that the emotional statement dimension and consequences factors in CIPQ-R were significantly negatively correlated with the total scores of QOLIE-31 and all the other factors it belongs to ( r=-0.202--0.527, P<0.05); the identity, timeline acute/chronic, and timeline cyclical factors in CIPQ-R were negatively correlated with the total score of QOLIE-31 ( r=-0.272--0.342, P<0.01); the treatment control and illness coherence factors in CIPQ-R were positively correlated with the total score of QOLIE-31 ( r=0.259, r=0.307, P<0.01). Multiple stepwise linear regression showed that the identity, timeline acute/chronic, consequences, timeline cyclical, and emotional representations factors were important factors influencing the quality of life of adults with epilepsy. Conclusion:Adult epileptics have certain negative illness perceptions.Among them, identity, timeline acute/chronic, consequences, timeline cyclical and emotional representations factors have significant effects on the quality of life of adult epileptics.

Objective:To explore the clinical technique and method of reconstruction large soft tissue defect of plantar by skin soft tissue dilatation combined with thigh cross leg flap to improve the weight-bearing function of plantar standing.Methods:From February 2015 to June 2019, 8 cases with plantar soft tissue defects were repaired with cross leg flap expanded from the healthy side of the thigh. Right foot in 4 cases, left foot in 3 cases and bilateral in 1 case. There were 5 males and 3 females. Ages ranged from 16 to 56, with an average age of 31. Clinical features: scar ulcer in 4 cases, bone-sticking scar in 4 cases, bone exposure in 1 case. After debridement, the area of skin and soft tissue defect was 9 cm×16 cm-12 cm×29 cm, the area of harvest expanded flap was 11 cm×18 cm-14 cm×30 cm. All flaps were fixed by cross-leg with pedicle. The pedicle division were performed one month after the skin flap transmission.Results:Of the 9 flaps transferred, 8 survived, 1 died in the distal part of the flap and was repaired by secondary expanded flap, and the donor site healed in linear scar stage. After 6-24 months follow-up, the plantar flap was in good shape and could walk with weight-bearing, without ulceration, and the protective tactile sensation was gradually restored in 12 months.Conclusions:The large-area soft tissue defect of plantar was repaired with the expanded cross leg flap in the healthy thigh, which also provide sufficient thickness of soft tissue to increase the ability of bearing weight and wearing resistance of the sole. It is a good alternative method to repair large area soft tissue defect of plantar under the condition of limited transfer of perforator flap and free flap.

Objective:To explore the clinical technique and method of reconstruction large soft tissue defect of plantar by skin soft tissue dilatation combined with thigh cross leg flap to improve the weight-bearing function of plantar standing.Methods:From February 2015 to June 2019, 8 cases with plantar soft tissue defects were repaired with cross leg flap expanded from the healthy side of the thigh. Right foot in 4 cases, left foot in 3 cases and bilateral in 1 case. There were 5 males and 3 females. Ages ranged from 16 to 56, with an average age of 31. Clinical features: scar ulcer in 4 cases, bone-sticking scar in 4 cases, bone exposure in 1 case. After debridement, the area of skin and soft tissue defect was 9 cm×16 cm-12 cm×29 cm, the area of harvest expanded flap was 11 cm×18 cm-14 cm×30 cm. All flaps were fixed by cross-leg with pedicle. The pedicle division were performed one month after the skin flap transmission.Results:Of the 9 flaps transferred, 8 survived, 1 died in the distal part of the flap and was repaired by secondary expanded flap, and the donor site healed in linear scar stage. After 6-24 months follow-up, the plantar flap was in good shape and could walk with weight-bearing, without ulceration, and the protective tactile sensation was gradually restored in 12 months.Conclusions:The large-area soft tissue defect of plantar was repaired with the expanded cross leg flap in the healthy thigh, which also provide sufficient thickness of soft tissue to increase the ability of bearing weight and wearing resistance of the sole. It is a good alternative method to repair large area soft tissue defect of plantar under the condition of limited transfer of perforator flap and free flap.

Ten patients with perianal necrotizing fasciitis were admitted to our department from June to December 2016. There were 8 men and 2 women among the patients, aged 42 to 69 years. Early and complete debridement surgery and comprehensive supportive treatment during perioperative period were carried out to quickly stabilize the patient′s overall condition, and wounds were sutured directly or repaired with autologous scalps and or adjacent local random flaps. After debridement, wound areas ranged from 10 cm×8 cm to 54 cm×21 cm, and area of the flap was about 8 cm×5 cm. The donor site of flap was sutured directly. After the operation, all skin grafts and the flap survived, and wounds of all patients healed. During follow-up of six months to one year, there was no recurrence of perianal necrotizing fasciitis, and functions of the involved lower extremities didn′t be influenced.