OBJECTIVETo develop a method for extracting cytoskeletons and cytoskeleton-associated proteins for proteomic analysis.

METHODSA subcellular sequential proteome extraction method was exploited. The extraction procedure was optimized and controlled according to observed cell morphology changes and one- and two-dimensional electrophoresis images. The extraction efficiency and selectivity were evaluated by Western blotting and mass spectrometry.

RESULTSFour extracted fractions clearly displayed distinct patterns. Western blotting detected the fraction-marker proteins FAK, integrin-β1, histone H1 and cytokeratin 19 only in their expected fractions. About 90% of the protein spots in the cytoskeleton fraction were identified by mass spectrometry as cytoskeleton and/or its associated proteins.

CONCLUSIONThe subcellular proteome sequential fractionation method facilitates the detection of proteins of low abundance and shows a high reproducibility and selectivity, and thus can serve as an ideal pre-fractionation method prior to two-dimensional electrophoresis.

Cytoskeleton ; chemistry ; Electrophoresis, Gel, Two-Dimensional ; methods ; Humans ; Proteome ; analysis ; Proteomics ; methods ; Subcellular Fractions

Objective:To investigate the outcome of epicanthoplasty combined with double eyelidplasty in different aesthetic forms of double eyelids (inner type and outer type) using Z-epicanthoplasty and inverted L method.Methods:The clinical data of patients with single eyelids and epicanthus were retrospectively analyzed from November 2021 to October 2022 in Gaoxin Hospital of the First Affiliated Hospital of Nanchang University. Patients were divided into inner type group and outer type group based on their aesthetic requirements for inner double eyelids. Both groups underwent Z-epicanthoplasty or inverted L method for epicanthus correction. Postoperative follow-up was conducted to observe the incision healing, complications, and postoperative effects of the two groups. The correction effects [ineffective, effective, and markedly effective, with the effective rate calculated as (the number of markedly effective cases + the number of effective cases)/total number of cases × 100%], scar scores, and patient satisfaction scores were collected for more than 12 months after surgery. Independent sample t-test was used for the scar scores and patient satisfaction scores, and chi-square test was used for the epicanthus correction effects. P<0.05 indicated a statistically significant difference. Results:A total of 76 patients were included. Among them, 41 patients in the inner type group (4 males and 37 females, with an average age of 22.3 years, ranging from 18 to 28 years) underwent 19 cases of Z-epicanthoplasty and 22 cases of inverted L method; 35 patients in the outer type group (3 males and 32 females, with an average age of 24.2 years, ranging from 19 to 30 years) underwent 20 cases of Z-epicanthoplasty and 15 cases of inverted L method. After 12 to 16 months of follow-up, with an average of 14 months, all incisions in both groups healed in the first stage, with no significant scar hyperplasia or other complications. Most of the double eyelid shapes are natural after the surgery, and the correction of epicanthus is obvious. Comparison of the treatment efficacy between Z-epicanthoplasty and inverted L method showed no significant difference [94.87% (37/39) vs. 94.59% (35/37), P>0.05]. In the inner type group, the postoperative scar score of the inverted L method was lower than that of the Z-epicanthoplasty [(6.81±0.66) vs. (7.31±0.88)], and the satisfaction score was better than that of the Z-epicanthoplasty [(0.86±0.63) vs. (1.73±0.65)], with statistically significant differences (both P<0.05). In the outer type group, the scar score of the inverted L method was higher than that of the Z-epicanthoplasty [(7.66±0.89) vs. (7.00±0.64)], and the satisfaction score was worse than that of the Z-epicanthoplasty [(1.80±0.86) vs. (1.00±0.56)], with statistically significant differences (both P<0.05). Conclusion:For patients requiring inner type aesthetic form of medial double eyelids, inverted L method can achieve less postoperative scars and higher postoperative satisfaction compared with Z-epicanthoplasty. For patients requiring "outer type" aesthetic form, Z-epicanthoplasty can achieve less postoperative scars and higher postoperative satisfaction compared with inverted L method.

Objective:To investigate the outcome of epicanthoplasty combined with double eyelidplasty in different aesthetic forms of double eyelids (inner type and outer type) using Z-epicanthoplasty and inverted L method.Methods:The clinical data of patients with single eyelids and epicanthus were retrospectively analyzed from November 2021 to October 2022 in Gaoxin Hospital of the First Affiliated Hospital of Nanchang University. Patients were divided into inner type group and outer type group based on their aesthetic requirements for inner double eyelids. Both groups underwent Z-epicanthoplasty or inverted L method for epicanthus correction. Postoperative follow-up was conducted to observe the incision healing, complications, and postoperative effects of the two groups. The correction effects [ineffective, effective, and markedly effective, with the effective rate calculated as (the number of markedly effective cases + the number of effective cases)/total number of cases × 100%], scar scores, and patient satisfaction scores were collected for more than 12 months after surgery. Independent sample t-test was used for the scar scores and patient satisfaction scores, and chi-square test was used for the epicanthus correction effects. P<0.05 indicated a statistically significant difference. Results:A total of 76 patients were included. Among them, 41 patients in the inner type group (4 males and 37 females, with an average age of 22.3 years, ranging from 18 to 28 years) underwent 19 cases of Z-epicanthoplasty and 22 cases of inverted L method; 35 patients in the outer type group (3 males and 32 females, with an average age of 24.2 years, ranging from 19 to 30 years) underwent 20 cases of Z-epicanthoplasty and 15 cases of inverted L method. After 12 to 16 months of follow-up, with an average of 14 months, all incisions in both groups healed in the first stage, with no significant scar hyperplasia or other complications. Most of the double eyelid shapes are natural after the surgery, and the correction of epicanthus is obvious. Comparison of the treatment efficacy between Z-epicanthoplasty and inverted L method showed no significant difference [94.87% (37/39) vs. 94.59% (35/37), P>0.05]. In the inner type group, the postoperative scar score of the inverted L method was lower than that of the Z-epicanthoplasty [(6.81±0.66) vs. (7.31±0.88)], and the satisfaction score was better than that of the Z-epicanthoplasty [(0.86±0.63) vs. (1.73±0.65)], with statistically significant differences (both P<0.05). In the outer type group, the scar score of the inverted L method was higher than that of the Z-epicanthoplasty [(7.66±0.89) vs. (7.00±0.64)], and the satisfaction score was worse than that of the Z-epicanthoplasty [(1.80±0.86) vs. (1.00±0.56)], with statistically significant differences (both P<0.05). Conclusion:For patients requiring inner type aesthetic form of medial double eyelids, inverted L method can achieve less postoperative scars and higher postoperative satisfaction compared with Z-epicanthoplasty. For patients requiring "outer type" aesthetic form, Z-epicanthoplasty can achieve less postoperative scars and higher postoperative satisfaction compared with inverted L method.

Objective To investigate the plasma levels of signal peptide-CUB epidermal growth fac-tor domain inclusion protein-1(SCUBE-1),asymmetric dimethylarginine(ADMA)and insulin-like growth factor binding protein-2(IGFBP-2)in elderly patients with acute pulmonary embol-ism(PE)and their relationship with risk stratification and poor prognosis.Methods A total of 117 elderly patients with acute PE admitted to our hospital from January 2022 to December 2023 were retrospectively enrolled in this study.With risk stratification,there were 41 patients of high-risk,46 of medium-risk and 30 of low-risk.According to their clinical outcomes,they were divided into good prognosis group(85 cases)and bad prognosis group(32 cases).The plasma levels of SCUBE-1,ADMA,and IGFBP-2 were measured using ELISA.Multivariate logistic regression analysis was applied to identify the risk factors for poor prognosis,and ROC curves were drawn to analyze the predictive value of plasma levels of SCUBE-1,ADM A and IGFBP-2 for poor prognosis of these patients.Results The levels of cTnI,BNP,D-dimer,SCUBE-1,ADMA and IGFBP-2 were significantly higher in the poor prognosis group than the good prognosis group(0.95±0.28μg/L vs 0.30±0.08 μg/L,2017.62±308.15 μg/L vs 538.14±102.73 μg/L,5.24±1.13 μg/L vs 1.63±0.27 μg/L,22.50±10.14 μg/L vs 9.25±4.36 μg/L,2.01±0.72 μmol/L vs 0.61±0.17μmol/L,468.16±90.51 μg/L vs 155.27±30.74 μg/L,P＜0.01).The high-risk patients had high-er plasma levels of SCUBE-1,ADMA and IGFBP-2 than the medium-and low-risk patients in or-der(all P＜0.01).Multivariate logistic regression analysis showed that elevated levels of cTnI,BNP,D-dimer,SCUBE-1,ADMA and IGFBP-2 were risk factors for death in acute PE patients(P＜0.05).ROC curve analysis indicated that the AUC value of SCUBE-1,ADMA and IGFBP-2 combined together in predicting death for acute PE patients was the highest(0.927,95％CI:0.864-0.993),with an accuracy of 93.1％.The plasma levels of SCUBE-1,ADMA and IGFBP-2 were positively correlated with the levels of cTnI,BNP and D-dimer in the elderly patients(P＜0.01).Conclusion For the elderly patients with acute PE,elevated plasma levels of SCUBE-1,ADMA and IGFBP-2 are associated with risk stratification and poor prognosis.Combination of these three indicators has certain predictive value for poor prognosis in the patients.