Objective To explore the characteristics and mechanism of phase separation between TAR DNA binding protein-43(TDP-43)and ubiquitin.Methods The TARDBP gene and its truncated genes were inserted into vectors to construct recombinant plasmids for expression and protein purification.The phase separation system of ubiquitin and TDP-43 was constructed in vitro.The characteristics of the droplets formed via liquid-liquid phase separation were observed by fluorescence microscopy.The plasmids of ubiquitin and TDP-43 were co-transfected into HEK293T cells to observe aggregates containing TDP-43 and ubiquitin and find out whether TDP-43 could be ubiquitinated.Results The GFP-8Ub,TDP-43 full-length(FL)and truncated proteins were purified.TDP-43 FL and C-terminal domain(CTD)proteins were able to form droplets via phase separation with ubiquitin.The droplets changed into solid-like aggregates after prolonged incubation.Insolvable aggregates containing TDP-43 and ubiquitin were formed.TDP-43 was ubiquitinated under stress conditions in HEK293T cells after being co-transfected with ubiquitin and TDP-43 recombinant plasmids.Conclusion TDP-43 undergoes co-phase separation with ubiquitin,mainly driven by the multivalent interaction between TDP-43′s CTD structural domain and ubiquitin.The droplets finally form aggregates with solid-like properties.Under stress conditions,especially when the protein homeostasis is disrupted,TDP-43 and ubiquitin form aggregates while TDP-43 is ubiquitinated.This study reveals the basic mechanism of TDP-43 co-phase separation with ubiquitin and liquid-solid transformation.

Acute skin failure (ASF) is an inevitable damage to the skin and subcutaneous tissue caused by hemodynamic instability and/or low perfusion. At present, there are some understandings and reports about adult ASF at home and abroad, but there are few reports about children's ASF. This article reviewed the definition, pathophysiological changes, risk factors, clinical manifestations, and management of children's ASF, and put forward suggestions in order to provide ideas for clinical diagnosis and treatment of children's ASF, and promote the further study of children's ASF.

Objective To investigate the expression of miR-30a-5p and miR-129-5p in the serum of patients with non-small cell lung cancer after EGFR targeted therapy and their relationship with the therapeutic effect.Methods A total of 186 patients with non-small cell lung cancer who received EGFR targeted therapy in our hospital from January 2020 to June 2022 were regarded as research objects.According to the efficacy of patients,they were separated into effective group(n=141)and ineffective group(n=45).The serum levels of miR-30a-5p and miR-129-5p were compared between the two groups;multivariate logistic regression was applied to analyze the factors influencing the efficacy of non-small cell lung cancer;receiver operating characteristic was applied to analyze the predictive value of serum miR-30a-5p,miR-129-5p levels for efficacy of non-small cell lung cancer.Results There were obvious differences in smoking history and TNM stage between the ineffective group and the effective group(P＜0.05).The expression levels of miR-30a-5p and miR-129-5p in the ineffective group were obviously lower than those in the effective group(P＜0.05).Multivariate logistic regression analysis showed that serum miR-30a-5p,miR-129-5p,smoking history,and TNM stage were all factors influencing the efficacy of non-small cell lung cancer(P＜0.05).Receiver operating characteristic showed that the AUC of the combination of serum miR-30a-5p and miR-129-5p to predict the efficacy of non-small cell lung cancer was 0.926,the sensitivity was 91.11％,and the specificity was 83.69％,which was better than their respective predictions(Z combination-miR-30a-5P=3.260,Zcombination-miR-129-5p=3.726,P=0.001,0.000).Conclusion The serum levels of miR-30a-5p and miR-129-5p in patients with non-small cell lung cancer who failed to respond to EGFR targeted therapy are obviously lower than those in the effective group;The combination of the two has a good predictive value for the efficacy of non-small cell lung cancer after EGFR targeted therapy.

Based on the content of medical professionalism and the theoretical model of knowledge, belief and practice, this paper used the methods of "inductive category construction" and "deductive category construction" to form the categories and analyze the text, taking the narrative materials of 112 "volunteer stars" outstanding individuals selected by the pre-medical student office of student affairs department of Peking University Health Science Center from 2019 to 2021 as the research subjects. The findings demonstrated that volunteer service had a positive effect on the cultivation of medical professionalism of medical freshmen, and played an effective role in guiding medical freshmen from the two dimensions of knowledge and belief, in which the role of medical voluntary service was more obvious. However, the breadth and depth of volunteer service in cultivating medical professionalism were limited. Based on this, this paper proposed that medical colleges and universities should emphasize voluntary service in the cultivation of medical professionalism, especially the development of medical voluntary service activities designed for medical students, and strengthen the cultivation effect by coordinating with other cultivation methods.

Objective:To evaluate the differential expression of blood routine in different types of infection and the diagnostic value of C-reactive protein (CRP), procalcitonin (PT), ferritin (SF) and lactate dehydrogenase (LDH) in bacterial and mycoplasma pneumonia and their early warning value in severe cases.Method:A total of 627 patients, including 176 cases of bacterial pneumonia, 275 cases of mycoplasma pneumonia, 176 cases of viral infection and 180 cases of normal control were collected from May 2018 to December 2019 in children′s Hospital Affiliated to Capital Institute of Pediatrics. The mycoplasma pneumonia group was divided into mild group (151 cases) and severe group (124 cases) according to the results of lavage fluid RNA-examination. All patients received completed blood routine test at the first day of admission, patients in bacteria group and Mycoplasma group received the examination of four inflammatory indicators. The Kruskal-Wallis test was used to analyze the differences in blood routine results between different infection groups, and the differences of inflammatory indexes between bacterial group and Mycoplasma mild and severe group. The receiver operating characteristic (ROC)-curve method was used to analyze the predictive value of inflammatory indexes between different infection groups.Results:There were significant differences in leukocyte count, neutrophil, lymphocyte and monocyte percentage between bacterial pneumonia, mycoplasma pneumonia, viral infection and normal control group ( P<0.05). The differences of four inflammatory indexes in bacterial group, mild Mycoplasma group and severe group were statistically significant ( P<0.05). The rest of the index (CRP, PCT, LDH, SF and white blood cell count) were P<0.05 (CRP: area under curve [AUC] 0.799; PCT: AUC 0.579; LDH: AUC 0.651; SF: AUC 0.854), in mild and severe mycoplasma group, except WBC, by ROC-curves analysis. The AUC value of the area under the curve of CRP and SF is high, and the sensitivity and specificity at the diagnostic critical point are high, which has great diagnostic value (CRP: diagnostic critical point 12.55 mg/L, sensitivity 0.719, specificity 0.755; SF: diagnostic critical point 176.02 μg/L, sensitivity 0.765, specificity 0.960). ROC curve results also showed that of PCT, White blood cell and neutrophil percentage had the diagnostic value in bacterial infection and mycoplasma infection, P<0.05 (PCT: AUC 0.658; leukocyte: AUC 0.804; neutrophil: AUC 0.630). Leukocyte count is the best differential index (diagnostic critical point 9.585×10 9/L, sensitivity 0.778, specificity 0.698), PCT has higher sensitivity at the diagnostic critical point of 0.55 μg/L, but the specificity is slightly lower (diagnostic critical point of 0.55 μg/L, sensitivity 0.862, specificity 0.366). Conclusions:PCT and leukocyte count can be used as the preferred inflammatory indexes to distinguish bacterial and mycoplasma infection. CRP, LDH, PCT and SF can be used as early warning indexes to evaluate severe mycoplasma infection.

Objective:To investigate the etiology, pathogenesis, clinical manifestations, and treatment of acute pancreatitis (AP) complicated with spleen infarction.Methods:A case of AP combined with splenic infarction was retrospectively analyzed and the literature was reviewed.Results:AP complicated with splenic infarction suggested that the patient′s condition was serious. The impairment of splenic immune function caused by splenic infarction may be an important cause of severe acute pancreatitis (SAP). Early detection and active treatment can obtain a good prognosis, and anticoagulant or antiplatelet aggregation treatment was not the key treatment.Conclusions:AP combined with splenic infarction may be an risk factor for SAP.

Objective:To investigate the current situation of critical thinking ability of nurses in Catheterization Room in China, and analyze its influencing factors, so as to provide reference for improving the critical thinking ability of nurses in Catheterization Room.Methods:In July 2020, the catheterization nurses from 154 hospitals, including Class Ⅲ Grade A, Class Ⅲ Grade B and Class Ⅱ Grade A, in 28 provincial capital cities of China were investigated with general information questionnaire and Chinese Critical Thinking Disposition Inventory (CTDI-CV) , by convenience sampling method. 422 questionnaires were collected in this survey, 408 were valid, and the effective rate was 96.68%.Results:The total score of critical thinking ability of the 408 catheterization nurses was (247.92±33.77) , which was at a low level as a whole. There were 361 nurses (88.48%) without positive critical thinking, 42 (10.29%) with positive critical thinking, and 5 (1.23%) with strong positive critical thinking. Binary Logistic regression analysis showed that nurse level, education background, professional title and position were the main influencing factors of critical thinking ability of nurses in Catheterization Room ( P<0.05) . Conclusions:The critical thinking attitude of nurses in Catheterization Room is generally negative, which is far from positive, and needs to be further improved. The nursing department should give more tendentious policies and support to the nurses in the Catheterization Room. At the same time, the nursing managers in the Catheterization Room should actively take measures to improve the critical thinking ability of nurses.

Objective:To explore the application of lecture-based learning (LBL) combined with problem-based learning (PBL) in the teaching of pathophysiology case analysis.Methods:The subjects were 62 preventive medicine majors of Grade 2015 and 68 stomatology majors of Grade 2015, and the course content was case analysis. PBL teaching was implemented in the preventive medicine major group and LBL+PBL teaching was implemented in the stomatology major group. Both groups received 6 class hours, among which the LBL+PBL teaching hours were allocated 2 class hours for LBL and 4 class hours for PBL. In addition, the questionnaire was used to evaluate the teaching effects between the two groups, and the descriptive analysis was used to evaluate relevant data lines.Results:Eleven students (17.742%) from the preventive medicine major group chose PBL teaching, while 38 students (92.647%) in the stomatology major group held a very positive attitude towards LBL+PBL teaching. It can be seen that the recognition degree of LBL+PBL teaching in the stomatology major group was higher than that of PBL teaching in the preventive medicine major group.Conclusion:Since students have received LBL teaching mode for a long time, it is difficult to accept and adapt to PBL teaching in terms of thinking and teamwork. Therefore, in the course of pathophysiology case analysis, LBL+PBL teaching shows a good effect, which is worthy of further exploration.

Objective: To explore the effect of high mobility group box-1 protein (HMGB1) on the balance of Th17/Treg in patients with immune thrombocytopenia (ITP). Methods: A total of 30 patients who were first diagnosed as ITP in the Fifth People's Hospital of Datong from July 2017 to April 2018 were selected as the case group, and another 30 healthy volunteers in the corresponding period were taken as the control group. The proportion of Th17 and Treg cells was detected by using flow cytometry, and the concentration of HMGB1, interleukin (IL)-17 and transforming growth factor β (TGF-β) in plasma was tested by using enzyme-linked immunosorbent assay (ELISA). Isolated peripheral blood mononuclear cells (PBMC) were cultured in vitro. After the treatment with recombinant human HMGB1 (rhHMGB1), real-time polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression changes in Treg cell transcription factor intracellular forkhead helix transcription factor 3 (Foxp3) and Th17 cell transcription factor retinoid related orphan receptor γt (RORγt). The differences of indicators in Treg cell transcription factor peripheral blood between the case group and the control group were compared, and the balance correlation between HMGB1 and Th17/Treg was analyzed. Results: Compared with the healthy control group, the proportion of Th17 cells and the expression level of HMGB1 and IL-17 in peripheral blood of ITP patients were increased (all P < 0.01), while the proportion of Treg cells and the level of TGF-β were decreased (all P < 0.01). The proportion of Th17 cells and the expression level of IL-17 and HMGB1 in peripheral blood of ITP patients were positively correlated with the concentration of HMGB1 (all P < 0.01); the proportion of Treg cells and the level of TGF-β were negatively correlated with the expression level of HMGB1 (all P < 0.01). In vitro experiments, the expression of intracellular RORγt mRNA was increased compared with the negative control group (1.50±0.24 vs. 0.93±0.22, t = 9.612, P < 0.01), and the expression of Foxp3 mRNA was decreased compared with the negative control group after the stimulation of PBMC by rhHMGB1 (0.72±0.19 vs. 1.08±0.18, t = 7.387, P < 0.01). Conclusion The high level of HMGB1 in the peripheral blood of ITP patients induces Th17/Treg imbalance and aggravates inflammatory reactions, which may be an important cause of ITP.

Objective To evaluate the clinical implementation value of bedside ultrasound monitoring of gastric residual volume in the guidance of enteral nutrition (EN) in severe patients with mechanical ventilation. Methods One hundred and forty-three patients undergoing nasal feeding EN and mechanical ventilation &ge; 72 hours in the Department of Intensive Care Unit (ICU) of Jinhua People's Hospital from May 2017 to August 2018 were enrolled, and they were randomly divided into two groups: a traditional control group (71 cases) and an ultrasound monitoring group (72 cases). The traditional control group applied 50 mL syringe to withdraw gastric juice from gastric tube, once every 4 hours, and then the EN protocol based on the gastric residual volume was adjusted; the ultrasound monitoring group used bedside ultrasound to monitor the gastric residual volume once every 4 hours, and then the EN protocol was adjusted accordingly. The changes of EN tolerance indexes e.g. abdominal distension, reflux vomiting, diarrhea, bowel sound hyperactivity or disappearance, intra-abdominal pressure, etc, the changes of incidence of ventilator-associated pneumonia (VAP), mechanical ventilation time and ICU hospitalization time in these two groups were observed. Results The incidences of abdominal distension, reflux vomiting, diarrhea, intestinal hyperactivity or disappearance of bowel sounds were significantly lower in the ultrasound monitoring group than those in the traditional control group [abdominal distension: 23.6% (17/72) vs. 28.2% (20/71), reflux vomiting: 18.1% (13/72) vs. 22.5% (16/71), diarrhea: 13.9% (10/72) vs. 14.1% (10/71), bowel sounds hyperactivity or disappearance: 9.7% (7/72) vs. 11.3% (8/71), all P < 0.05], the intra-abdominal pressure was obviously lower in the ultrasound monitoring group than that in the traditional control group [mmHg (1 mmHg = 0.133 kPa): 8.9±5.6 vs. 9.6±6.1, P < 0.05], and the incidence of VAP was significantly lower in the ultrasound monitoring group than that in the traditional control group [19.7% (14/72) vs. 23.9% (17/71), P < 0.05], the mechanical ventilation time and ICU hospitalization time were also significantly shorter in the ultrasound monitoring group than those in the traditional control group [mechanical ventilation time (days): 10.1±3.1 vs. 12.2±3.4, ICU hospitalization time (days): 16.8±4.2 vs. 20.6±4.8, all P < 0.05]. Conclusion The bedside ultrasound monitoring gastric residual volume can improve EN tolerance, reduce VAP incidence, and shorten the mechanical ventilation time and ICU hospitalization time in patients with mechanical ventilation.