Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.

Objective Network pharmacology and molecular docking and molecular dynamics techniques were used to investigate the mechanism of action of Alhagi sparsifolia Shap.in the treatment of sepsis and to perform animal experimental verification.Methods First,we screened the effective ingredients and their action targets of Alhagi sparsifolia Shap.,meanwhile,screened relevant action targets for the treatment of sepsis,constructed a protein interaction(PPI)network,and performed topology analysis to draw a TCM disease target network diagram.Second,Kyoto Encyclopedia of genes and genomes enrichment analysis was performed for core targets in the network diagram,along with gene ontology functional enrichment analysis.This was followed by molecular docking and molecular dynamics simulation experiment validation of the core targets.Finally,mice were used for the verification of animal experiments.Results Thirty active components of Alhagi sparsifolia Shap.were screened out,and the top 5 ranked by degree value were quercetin,(-)-epigallocatechin,(-)-Epigallocatechin Gallate,genistein,kaempferol and epigallocatechin with 196 action targets;2144 disease-related targets for sepsis,105 targets for Alhagi sparsifolia Shap.-sepsis intersection,and the core targets were TNF,IL-6,AKT1,VEGFA,CASP3,IL-1β Et al.PI3K-Akt,TNF,HIF-1,AGE-RAGE,IL-17 and other signaling pathways are involved to mediate inflammatory responses,apoptosis and other biological processes to exert therapeutic effects on sepsis.Molecular docking results showed that camelina flavanoids bound equally well to each key target,among which the conformations with the lowest binding energy were(-)-Epigallocatechin Gallate-IL-6 and quercetin-IL-6.Molecular dynamics simulations were performed on the two pairs of complexes,and the results indicated that the stable binding could be achieved through a combination of electrostatic,van der Waals potential,and hydrogen bonding interactions.Animal experiments confirmed that Alhagi sparsifolia Shap.could inhibit the activation of PI3K/Akt signaling pathway,decrease the protein expression of Caspase-3,VEGF and reduced peripheral blood inflammatory factors secretion of TNF-α、IL-1βand IL-6,alleviating inflammatory injury in tissues and organs.Conclusion The therapeutic effect of Alhagi sparsifolia Shap.on sepsis is achieved through multi biological processes,multi targets,and multi pathways.It provides a certain theoretical basis for the clinical application of camel spines as well as sepsis treatment.

Objective:To establish a core competence evaluation index system for thoracic specialist nurses.Methods:From April to September 2022, based on the iceberg model of post competence, the primary evaluation index system of core competence of thoracic specialist nurses was developed through literature review and semi-structured interview. The core competence evaluation index system for thoracic specialist nurses was determined through three rounds of letter consultation with 20 experts by Delphi method.Results:The effective recovery rates of the three rounds of expert inquiry questionnaires were 100.0% (20/20), 95.0% (19/20) and 100.0% (19/19), respectively. The expert authority coefficients were 0.925, 0.958 and 0.968, and the Kendall's concordance coefficients were 0.270, 0.235 and 0.184, respectively ( P<0.01). Finally, a core competency evaluation index system for thoracic specialist nurses was formed, including 6 first level indicators, 18 second level indicators and 68 third level indicators. Conclusions:The core competency evaluation index system for thoracic specialist nurses constructed based on post competence in this study is certain scientific, reliable and practical, which can provide a basis for the training and assessment of thoracic specialist nurses.

Lower extremity deep vein thrombosis (DVT) is one of the main complications in patients with traumatic fractures, and for severe patients, the DVT can even affect arterial blood supply, resulting in insufficient limb blood supply. If the thrombus breaks off, pulmonary embolism may occur, with a high mortality. The treatment and rehabilitation strategies of thrombosis in patients with lower extremity fractures have its particularity. DVT in traumatic fractures patients has attracted extensive attention and been largely studied, and the measures for prevention and treatment of DVT are constantly developing. In recent years, a series of thrombosis prevention and treatment guidelines have been updated at home and abroad, but there are still many doubts about the prevention and treatment of DVT in patients with different traumatic fractures. Accordingly, on the basis of summarizing the latest evidence-based medical evidence at home and abroad and the clinical experience of the majority of experts, the authors summarize the clinical treatment and prevention protocols for DVT in patients with traumatic fractures, and make this consensus on the examination and assessment, treatment, prevention and preventive measures for DVT in patients with different fractures so as to provide a practicable approach suitable for China ′s national conditions and improve the prognosis and the life quality of patients.

Objective:To investigate the effects of mitochondrial arginyl-tRNA synthase (RARS2) on cell proliferation, invasion, migration and chemotherapy resistance of pancreatic cancer.Methods:Human pancreatic cancer cell lines AsPC-1 and PANC-1 were divided into negative control group, RARS2 interference group-1, RARS2 interference group-2, RARS2 overexpression control group and RARS2 overexpression group. Cell proliferation and sensitivity to gemcitabine were detected by CCK-8 assay, and cell invasion and migration were detected by Transwell assay. Western blot was used to detect the expression of RARS2 under different concentrations and different times of gemcitabine treatment. Western blot and PCR were used to detect the expression of RARS2 in gemcitabine-resistant AsPC cell.Results:Inhibition of RARS2 expression in AsPC-1 and PANC-1 cells significantly inhibited cell proliferation and enhanced sensitivity of gemcitabine to chemotherapy. Overexpression of RARS2 enhanced cell proliferation and decreased sensitivity to gemcitabine. In AsPC-1 cells, the number of migrated cells (100×) in negative control group, RARS2 interference group-1, RARS2 interference group-2, RARS2 overexpression control group and RARS2 overexpression group were (586.7±37.4) cells/field, (195.7±18.6) cells/field, (237.0±17.1) cells/field, (157.7±19.1) cells/field, (456.0±23.1) cells/field, the number of invasive cells were (87.7±13.2) cells/field, (24.7±6.5) cells/field, (31.7±6.1) cells/field, (29.3±4.5) cells/field, (94.3±9.3) cells/field, respectively. The migration and invasion ability of cells were decreased after the expression of RARS2 was decreased, and the migration and invasion ability of cells were enhanced after the expression of RARS2 was increased. PCR and Western blot assay showed that RARS2 expression in the gemcitabine-resistant AsPC-1 was higher than that in the common cell line. In AsPC-1 cells, the expression of RARS2 increased with increasing gemcitabine concentration and treatment time.Conclusion:RARS2 promotes cell proliferation, invasion, migration and chemoresistance of pancreatic cancer, and expression of RARS2 is positively correlated with gemcitabine concentration and treatment time.

Objective: To explore the protective effects of anthrahydroquinone-2,6-disulfonate (AH 2 QDS) on the kidneys of paraquat (PQ) poisoned rats via the apelin-APJ pathway. Methods: Male Sprague Dawley rats were divided into four experimental groups: control, PQ, PQ+sivelestat, and PQ+AH 2 QDS. The PQ+sivelestat group served as the positive control group. The model of poisoning was established via intragastric treatment with a 20% PQ pesticide solution at 200 mg/kg. Two hours after poisoning, the PQ+sivelestat group was treated with sivelestat, while the PQ+AH 2 QDS group was given AH 2 QDS. Six rats were selected from each group on the first, third, and seventh days after poisoning and dissected after anesthesia. The PQ content of the kidneys was measured using the sodium disulfite method. Hematoxylin-eosin staining of renal tissues was performed to detect pathological changes. Apelin expression in the renal tissues was detected using immunofluorescence. Western blotting was used to detect the expression levels of the following proteins in the kidney tissues: IL-6, TNF-α, apelin-APJ (the apelin-Angiotensin receptor), NF-κB p65, caspase-1, caspase-8, glucose-regulated protein 78 (GRP78), and the C/EBP homologous protein (CHOP). In in vitro study, a PQ toxicity model was established using human tubular epithelial cells treated with standard PQ. Twenty-four hours after poisoning, sivelestat and AH 2 QDS were administered. The levels of oxidative stress in human renal tubular epithelial cells were assessed using a reactive oxygen species fluorescence probe. Results: The PQ content in the kidney tissues of the PQ group was higher than that of the PQ+AH 2 QDS group. Hematoxylin-eosin staining showed extensive hemorrhage and congestion in the renal parenchyma of the PQ group. Vacuolar degeneration of the renal tubule epithelial cells, deposition of crescent-like red staining material in renal follicles, infiltration by a few inflammatory cells, and a small number of cast formation were also observed. However, these pathological changes were less severe in the PQ+sivelestat group and the PQ+AH 2 QDS group (P<0.05). On the third day after poisoning, immunofluorescence assay showed that the level of apelin in the renal tissues was significantly higher in the PQ+AH 2 QDS group than in the PQ group. Western blotting analysis results showed that IL-6, TNF-α, NF-κB p65, caspase-1, caspase-8, GRP78, and CHOP protein levels in the PQ group were higher than in the PQ+AH 2 QDS group (P<0.05). The expression of apelin-APJ proteins in the PQ+AH 2 QDS group was higher than in the PQ+sivelestat and PQ groups (P<0.05); this difference was significant on Day 3 and Day 7. The level of oxidative stress in the renal tubular epithelial cells of the PQ+AH 2 QDS group and the PQ+sivelestat group was significantly lower than in the PQ group (P<0.05). Conclusions: This study confirms that AH 2 QDS has a protective effect on PQ-poisoned kidneys and its positive effect is superior to that of sivelestat. The mechanism of the protective effects of AH 2 QDS may be linked to reduction in cellular oxidative stress, PQ content of renal tissue, inflammatory injury, endoplasmic reticulum stress, and apoptosis. AH 2 QDS may play a role in the treatment of PQ poisoning by upregulating the expression of the apelin-APJ.

With the development of modern imaging technology and improvement of physical examination consciousness of people, the incidence of pancreatic cystic neoplasm (PCN) has increased significantly. The overall prognosis of PCN is good, and great progress in surgical techniques and multidisciplinary teamwork has been made in pancreatic surgery. Although many guidelines have been released at home and abroad, there are controversies in the differential diagnosis, operative indications, surgical procedure, and the follow-up strategies toward patients with PCN. There are also significant differences in the level of diagnosis and treatment of medical centers, which brings certain challenges to the standardized diagnosis and treatment of PCN. Based on the latest clinical guidelines and new developments in clinical researches, the authors summarize the controversies of different guidelines in the diagnosis, treatment and follow-up of PCN in order to further improve the standardization of the diagnosis and treatment.

Pancreatic cancer is extremely malignant and has poor prognosis. Surgery is the only way for the therapy. In recent years, minimally invasive surgery has been developed rapidly in pancreatic surgery. However, pancreatic cancer is often accompanied with vascular aggression and inflammation, which greatly increasing the difficulty of minimally invasive surgery procedure. Its feasibility, safety and radical tumor cure are still controversial. In this article, combined with the existing guidelines and the latest clinical researches, we compared the application of minimally invasive and open surgery in pancreatic cancer and summarized the development status and controversy of minimally invasive treatment of pancreatic cancer. Surgeons should strictly grasp the minimally invasive surgical adaptation certificate, and further carry out clinical research on minimally invasive treatment of pancreatic cancer, so as to standardize and enhance the development of minimally invasive pancreatic surgery in China.

Objective:To determine the expression of the E2F6 transcription factor in human malignant melanoma tissues and cell lines, and to evaluate the effect of E2F6 on proliferation, migration and invasion of a malignant melanoma cell line A375.Methods:Frozen tissues and paraffin-embedded tissue sections were collected from 50 cases of cutaneous malignant melanoma and 30 cases of pigmented nevus in Department of Dermatology, Chongqing Traditional Chinese Medicine Hospital from January 2012 to December 2017. Quantitative reverse transcription-PCR (qRT-PCR) was performed to determine the mRNA expression of E2F6 in the malignant melanoma and pigmented nevus tissues, as well as in 7 malignant melanoma cell lines (HM, A375, WM451, WM35, SK-MEL-1, Hs-695T and MDA-MB-435s) and pigmented nevus cells, and immunohistochemical study and Western blot analysis were conducted to determine the protein expression of E2F6 and β-catenin in the malignant melanoma tissues. An E2F6-inhibiting plasmid and a control plasmid were separately transfected into A375 cells by using a liposome-mediated transfection method, and the E2F6 gene-knockdown efficiency was verified by qRT-PCR and Western blot analysis. Cell counting kit-8 (CCK8) assay, soft-agar plate cloning assay, Transwell migration and invasion assays and 3D cell culture assay were conducted to evaluate the effect of E2F6 gene knockdown on the proliferation, migration and invasion of A375 cells, flow cytometry was performed to detect the cell cycle and apoptosis rate, and Western blot analysis was conducted to determine the protein expression of total β-catenin, activated β-catenin, c-Myc and cyclin D1. The comparison between two groups was carried out by t test, the comparison among several groups by one-way analysis of variance, and multiple comparisons by least significant difference t test; Pearson correlation coefficient was used to analyze the correlation between E2F6 and β-catenin expression in cutaneous malignant melanoma. Results:The E2F6 mRNA expression was significantly higher in the 7 malignant melanoma cell lines than in the pigmented nevus cells (all P < 0.001). qRT-PCR showed that the relative mRNA expression of E2F6 was significantly higher in the cutaneous malignant melanoma tissues (0.000 55 ± 0.000 17) than in the pigmented nevus tissues (0.000 18 ± 0.000 09, t = 3.22, P < 0.001). Both the immunohistochemical study and Western blot analysis showed significantly increased E2F6 protein expression, but decreased β-catenin protein expression in the cutaneous malignant melanoma tissues compared with the pigmented nevus tissues (all P < 0.001). Correlation analysis showed that E2F6 protein expression was negatively correlated with β-catenin expression in the malignant melanoma tissues (immunohistochemical study: r = -0.56, Western blot analysis: r = -0.63, both P < 0.01). After knockdown of the E2F6 gene in A375 cells, the mRNA and protein expression of E2F6 was significantly lower in the E2F6 inhibition group than in the control group ( t = 3.38, 2.76 respectively, both P < 0.001). CCK8 assay showed that the cellular proliferative ability was significantly lower in the E2F6 inhibition group than in the control group ( t = 4.58, P < 0.01) 48 hours after transfection; soft-agar plate cloning assay showed that the colony-formation ratio was significantly lower in the E2F6 inhibition group than in the control group ( t = 2.26, P < 0.001) ; Transwell migration and invasion assays showed that the number of cells crossing the chamber was significantly lower in the E2F6 inhibition group (165 ± 23, 96 ± 11 respectively) than in the control group (376 ± 22, 315 ± 31, t = 3.14, 2.12, respectively, both P < 0.01) ; 3D cell culture assay showed that the cell morphology markedly changed, and the invasive pseudopodia disappeared in the E2F6 inhibition group. Flow cytometry revealed that the proportion of cells at G0-G1 phase and apoptosis rate were significantly higher in the E2F6 inhibition group than in the control group (both P < 0.001). Western blot analysis showed significantly decreased protein expression of β-catenin, activated β-catenin and its downstream target proteins c-Myc and cyclin D1, but significantly increased protein expression of P21 in the E2F6 inhibition group compared with the control group (all P < 0.001) ; additionally, the E2F6 inhibition group showed significantly decreased protein expression of epithelial-mesenchymal transition-related molecules vimentin and N-cadherin, but significantly increased expression of E-cadherin compared with the control group (all P < 0.001) . Conclusions:The E2F6 transcription factor is highly expressed in malignant melanoma. Knockdown of the E2F6 gene in A375 cells can inhibit cell proliferation, migration and invasion by antagonizing the β-catenin signaling pathway.

Since December 2019, novel coronavirus pneumonia (NCP) has been reported in Wuhan, Hubei Province, and spreads rapidly to all through Hubei Province and even to the whole country. The virus is 2019 novel coronavirus (2019-nCoV), never been seen previously in human, but all the population is generally susceptible. The virus spreads through many ways and is highly infectious, which brings great difficulties to the prevention and control of NCP. Based on the needs of orthopedic trauma patients for emergency surgery and review of the latest NCP diagnosis and treatment strategy and the latest principles and principles of evidence-based medicine in traumatic orthopedics, the authors put forward this expert consensus to systematically standardize the clinical pathway and protective measures of emergency surgery for orthopedic trauma patients during prevention and control of NCP and provide reference for the emergency surgical treatment of orthopedic trauma patients in hospitals at all levels.