Background Pneumoconiosis is the most serious occupational disease in China, among which silicosis accounts for more than 50%. microRNA (miRNA) plays an important role in the occurrence process of silicosis fibrosis, but the mechanism of it has not been fully clarified yet. Objective To explore the molecular mechanism by which miR-411-3p modulates the ubiquitination degradation of SMAD specific E3 ubiquitin protein ligase (SMURF) 2/Smad7, thereby suppressing epithelial-mesenchymal transition (EMT) in mouse alveolar type II epithelial cells and counteracting silica-induced pulmonary fibrosis. Methods Twenty-four 8-week-old SPF male C57BL/6J mice were randomly divided into four groups: Control group, silica group, silica +miR-411-3p agomir-NC group, and silica +miR-411-3p agomir group, with 6 mice in each group. Silicosis model was prepared by a one-time bronchial infusion of silicon dioxide (SiO₂) (200 mg·mL^-1, 50 μL). In vitro MLE-12 cells were divided into (1) control group and SiO₂ group, (2) SiO₂+negative control siRNA (siRNA-NC) group and SiO₂+Smurf2 gene silencing (si-Smurf2) group, (3) SiO₂+solvent (DMSO) group and SiO₂+protease inhibitor (MG132) group, (4) mutant sequence plasmid (Mut)+miR-411-3p mimic control (miR-NC) group, Mut+miR-411-3p mimic group, wild sequence plasmid (Wt)+miR-NC group, and Wt+miR-411-3p mimic group, (5) SiO₂+miR-NC group and SiO₂+miR-411-3p mimic group. The pathological morphology and collagen deposition of lung tissue were observed after staining. Detection of miR-411-3p and proteins was conducted by real-time fluorescent quantitative PCR and Western blot. The binding of SMURF2 to Smad7 protein and Smad7 to ubiquitin (Ub) were detected by co-immunoprecipitation (Co-IP) method. Dual-luciferase reporter gene assay was adopted to verify the regulatory effect of miR-411-3p on Smurf2. Results In the SiO₂-induced MLE-12 cells, compared to the control group, the SiO₂-treated group showed significantly upregulated expressions of N-cadherin (N-Cad), collagen I (CoL I), SMURF2, transforming growth factor-β1 (TGF-β1), and phosphorylated Smad2/3 (p-Smad2/3). In contrast, the expressions of E-cadherin (E-Cad), Smad7, and miR-411-3p were significantly downregulated (P<0.05). The dual-luciferase reporter gene assay revealed a regulatory effect of miR-411-3p on Smurf2 (P<0.05). Meanwhile, in the MLE-12 cells induced by SiO₂, the miR-411-3p mimic down-regulated the protein expressions of SMURF2, N-Cad, CoL I, TGF-β1, and p-Smad2/3, while up-regulated the protein expressions of E-Cad and Smad7 (P<0.05). The silenced Smurf2 gene inhibited the expressions of N-Cad, CoL I, and p-Smad2/3 proteins, while promoted the expressions of E-Cad and Smad7 proteins in the MLE-12 cells (P<0.05). The Co-IP results showed that the binding of SMURF2 to Smad7 was enhanced, and the ubiquitin binding ability of Smad7 was enhanced in the SiO₂ group. In the lung tissue of mice, the results of pathological observation with hematoxylin-eosin (HE) and sirius red (VG) staining showed that compared with the agomir-NC, the lesion was relieved in the lung tissue of the miR-411-3p agomir group. Meanwhile, the expressions of SMURF2, N-Cad, CoL I, TGF-β1, and p-Smad2/3 were significantly down-regulated, while the expressions of E-Cad and Smad7 were significantly up-regulated (P<0.05). Conclusion MiR-411-3p alleviates the EMT of alveolar type II epithelial cells and antagonizes silicosis fibrosis progression in mice by inhibiting SMURF2-mediated ubiquitination and degradation of Smad7.

Objective:To evaluate the prognostic value of sepsis-induced coagulopathy (SIC) in patients with sepsis.Methods:From January 2019 to December 2021, patients with sepsis admitted to the Intensive Care Unit of our hospital were retrospectively classified into the SIC group and non-SIC group according to SIC diagnostic criteria. The baseline clinical data, severity score, total length of hospital stay, length of ICU stay and 28-day survival were compared between the two groups. Kaplan-Meier was used to compare the 28-day survival of patients with sepsis between the two groups. Cox proportional hazard regression model was employed to analyze the risk factors of prognosis in patients with sepsis.Results:Totally 274 patients with sepsis were included in the analysis, including 139 patients in the SIC group and 135 patients in the non-SIC group. The two groups were compared in the perspectives of the Platelet count (PLT), prothrombin time (PT) , procalcitonin (PCT), D dimer, hematocrit, red blood cell distribution width, hemoglobin, acute kidney injury (AKI), the use of continuous renal replacement treatment (CRRT), the use of vasoactive drugs, sequential organ failure assessment (SOFA) score, acute physiology and chronic health evaluation (APACHEⅡ) score were compared between the two groups and the difference were statistically different (all P<0.05). Kaplan-Meier analysis showed that the 28-day mortality rate in the SIC group was significantly higher than that in the non-SIC group (32.4% vs. 14.1%, P<0.05). COX proportional hazard model showed that SIC score ( HR= 2.17, 95% CI: 1.15-3.91, P<0.05), APACHEⅡ score ( HR= 1.13, 95% CI: 1.09-1.17, P<0.05) and the use of vasoactive drugs ( HR=3.66, 95% CI: 1.53-8.75, P<0.05) were independent influencing factors for 28-day death in patients with sepsis. Conclusions:Patients with sepsis and SIC have more severe disease and increased mortality risk. SIC score exhibits good clinical value in predicting the prognosis of patients with sepsis.

OBJECTIVE： To optimize the ethanol extraction technology of total alkaloids from Shuanghu capsules. METHODS： Using dendrobine as control， the contents of total alkaloids from Dendrobium nobile and Dendrobium officinale in Shuanghu capsules were determined by acidic dyes colorimetry. Using comprehensive scores calculated by the yield of the extract and the contents of total alkaloids as evaluation indexes， the effects of soaking time， ethanol volume fraction， extraction time， solid-liquid ratio and extraction times were investigated with single factor tests. L9（34） orthogonal test was used to optimize ethanol volume fraction， extraction time， solid-liquid ratio and extraction times according to the results of single factor test. The optimized technology was validated. RESULTS： The linear range of dendrobine were 4.16-14.56 μg/mL （r＝0.999 2）. RSDs of repeatability and precision tests were all lower than 5％. Average recovery tests were 93.01％ （RSD＝1.97％， n＝6）. The optimal ethanol extraction technology included soaking for 12 h， ethanol volume fraction of 70％， solid-liquid ratio of 1 ∶ 12 （g/mL）， extracting for 28 min， extracting 3 times. Results of validation test showed that the average yield of extract was 12.80％ （RSD＝4.39％， n＝3）， and the content of alkaloids was 0.359 0 mg/g（RSD＝0.66％， n＝3）. CONCLUSIONS： Established acidic dyes colorimetry is simple， precise and accurate， which can be used for the content determination of total alkaloids. The optimized ethanol extraction technology is stable and feasible， and can be used for the extraction of total alkaloids from Shuanghu capsules.

OBJECTIVE： To optimize the water extraction technology of total polysaccharide of Shuanghu capsules. METHODS： The total alkaloid was firstly extracted from Dendrobium nobile and Dendrobium officinale mixture of Shuanghu capsules with ethanol， and then total polysaccharide was extracted with water. Using glucose as control， total polysaccharide was treated with phenol-sulfuric acid method and its content was determined at 488 nm. Using comprehensive score calculated with the yield of the extract and the content of total polysaccharide as index， the effects of material-liquid ratio， extraction temperature， extraction time and times on the extraction were investigated by single factor test. Then L9（34） orthogonal test was used to optimize solid-liquid ratio，extraction temperature，extraction time and extraction times according to the results of single factor test. The optimized technology was validated. RESULTS： The linear range of glucose were 0.041 4-0.207 0 mg/mL（r＝0.999 9）. RSDs of intra-day and inter-day ranged 3.61％-8.24％ （n＝3，n＝5）， and RSD of repeatability test was 1.49％ （n＝6）. Average recovery rate was 98.65％（RSD＝1.45％，n＝6）. The optimal water extraction technology included solid-liquid ratio of 1 ∶ 25（g/mL），extraction temperature of 100 ℃，extracting for 90 min， extracting once. Results of validation tests showed that average content of total polysaccharide was 379.292 8 mg/g （RSD＝1.93％，n＝3） and average yield of the extract was 22.75％（RSD＝2.41％，n＝3）. CONCLUSIONS： Established phenol-sulphuric acid method is simple， precise and accurate. The optimal water extraction technology is stable and feasible， which can be used for the extraction of total polysaccharides from Shuanghu capsules.