Objective:To evaluate the effect of Alda-1 on ferroptosis in cardiomyocytes after cardiac arrest and cardiopulmonary resuscitation in swine.Methods:Twenty-two healthy male white swine, weighing 35-43 kg, were divided into 3 groups using a random number table method: sham operation group (group S, n=6), cardiac arrest-cardiopulmonary resuscitation group (group CA-CPR, n=8) and Alda-1 group ( n=8). The animals only underwent the general preparation in group S, and the swine model of cardiac arrest and cardiopulmonary resuscitation was developed by 8 min of electrically induced cardiac arrest through the pacing catheter in the right ventricle followed by 8 min of cardiopulmonary resuscitation in CA-CPR and Alda-1 groups.Alda-1 0.88 mg/kg was intravenously injected at 5 min after resuscitation in group Alda-1, and the equal volume of vehicle was administered instead in the other two groups.Stroke volume (SV) and global ejection fraction (GEF) were measured using PiCCO before developing the model and at 1, 2 and 4 h after resuscitation (T 0-3). Venous blood samples were collected from the femoral vein to measure the concentrations of serum cardiac troponin (cTnI) by enzyme-linked immunosorbent assay at T 0-3, and at 24 h after resuscitation (T 4). The animals were then sacrificed, and myocardial tissues in the left ventricle were harvested to measure the expression of acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) (by Western blot), iron deposition (by Prussian blue staining), 4-hydroxy-2-nonenal (4-HNE) content (by enzyme-linked immunosorbent assay), and malondialdehyde (MDA) and glutathione (GSH) contents (by colorimetry). Results:Compared with group S, SV and GEF were significantly decreased at T 1-3, the serum concentrations of cTnI were increased at T 1-4, myocardial ACSL4 expression was up-regulated, GPX4 expression was down-regulated, iron deposition and contents of 4-HNE and MDA were increased, and the content of GSH was decreased in CA-CPR and Alda-1 groups ( P<0.05). Compared with group CA-CPR, SV and GEF were significantly increased at T 2-3, the serum concentrations of cTnI were decreased at T 3-4, myocardial ACSL4 expression was down-regulated, GPX4 expression was up-regulated, iron deposition and contents of 4-HNE and MDA were decreased, and the content of GSH was increased in group Alda-1 ( P<0.05). Conclusions:Alda-1 can alleviate myocardial injury after cardiac arrest and cardiopulmonary resuscitation in swine and further improve cardiac dysfunction, and the mechanism may be related to inhibition of cell ferroptosis.

Na +-K +-2Cl-cotransporter 1 (NKCC1) is highly expressed in malignant gliomas,which is closely related to the degree of malignancy.NKCC1 protein has a vital function in the volume regulation of glioma cells.NKCC1 allows glioma cells to transform its volume freely,migrating through the narrow extracellular space to achieve distant metastases.There is also close relationship between NKCC1 and tumor cytoskeleton regulation.In addition,NKCC1 is closely associated with cell cycle,nerve activity and other biological functions.In conclusion,NKCC1 plays an important role in gliomas.

OBJECTIVE: To investigate whether the acetaldehyde dehydrogenase 2 specific activator, Alda-1, can alleviate brain injury after cardiopulmonary resuscitation (CPR) by inhibiting cell ferroptosis mediated by acyl-CoA synthetase long-chain family member 4/glutathione peroxidase 4 (ACSL4/GPx4) pathway in swine. METHODS: Twenty-two conventional healthy male white swine were divided into Sham group (n = 6), CPR model group (n = 8), and Alda-1 intervention group (CPR+Alda-1 group, n = 8) using a random number table. The swine model of CPR was reproduced by 8 minutes of cardiac arrest induced by ventricular fibrillation through electrical stimulation in the right ventricle followed by 8 minutes of CPR. The Sham group only experienced general preparation. A dose of 0.88 mg/kg of Alda-1 was intravenously injected at 5 minutes after resuscitation in the CPR+Alda-1 group. The same volume of saline was infused in the Sham and CPR model groups. Blood samples were collected from the femoral vein before modeling and 1, 2, 4, 24 hours after resuscitation, and the serum levels of neuron specific enolase (NSE) and S100 β protein were determined by enzyme-linked immunosorbent assay (ELISA). At 24 hours after resuscitation, the status of neurologic function was evaluated by neurological deficit score (NDS). Thereafter, the animals were sacrificed, and brain cortex was harvested to measure iron deposition by Prussian blue staining, malondialdehyde (MDA) and glutathione (GSH) contents by colorimetry, and ACSL4 and GPx4 protein expressions by Western blotting. RESULTS: Compared with the Sham group, the serum levels of NSE and S100β after resuscitation were gradually increased over time, and the NDS score was significantly increased, brain cortical iron deposition and MDA content were significantly increased, GSH content and GPx4 protein expression in brain cortical were significantly decreased, and ACSL4 protein expression was significantly increased at 24 hours after resuscitation in the CPR model and CPR+Alda-1 groups, which indicated that cell ferroptosis occurred in the brain cortex, and the ACSL4/GPx4 pathway participated in this process of cell ferroptosis. Compared with the CPR model group, the serum levels of NSE and S100 β starting 2 hours after resuscitation were significantly decreased in the CPR+Alda-1 group [NSE (μg/L): 24.1±2.4 vs. 28.2±2.1, S100 β (ng/L): 2 279±169 vs. 2 620±241, both P < 0.05]; at 24 hours after resuscitation, the NDS score and brain cortical iron deposition and MDA content were significantly decreased [NDS score: 120±44 vs. 207±68, iron deposition: (2.61±0.36)% vs. (6.31±1.66)%, MDA (μmol/g): 2.93±0.30 vs. 3.68±0.29, all P < 0.05], brain cortical GSH content and GPx4 expression in brain cortical was significantly increased [GSH (mg/g): 4.59±0.63 vs. 3.51±0.56, GPx4 protein (GPx4/GAPDH): 0.54±0.14 vs. 0.21±0.08, both P < 0.05], and ACSL4 protein expression was significantly decreased (ACSL4/GAPDH: 0.46±0.08 vs. 0.85±0.13, P < 0.05), which indicated that Alda-1 might alleviate brain cortical cell ferroptosis through regulating ACSL4/GPx4 pathway. CONCLUSIONS Alda-1 can reduce brain injury after CPR in swine, which may be related to the inhibition of ACSL4/GPx4 pathway mediated ferroptosis.
Male ; Animals ; Swine ; Phospholipid Hydroperoxide Glutathione Peroxidase ; Ferroptosis ; Brain Injuries ; Glutathione ; Cardiopulmonary Resuscitation ; Ligases ; Iron

Objective:To study the effect of duodenal papillary morphology on selective biliary cannulation (SBC).Methods:Data of 912 patients with biliopancreatic diseases treated with endoscopic retrograde cholangiopancreatography (ERCP) at the Department of Gastroenterology, Affiliated Hospital of Northwest Minzu University from January 2018 to January 2020 were retrospectively analyzed. Duodenal papillary morphology of patients was classified into 4 types by using Haraldsson's endoscopic classification. The success rate of SBC, the pre-incision rate, the difficult intubation rate and the incidence of post-ERCP pancreatitis (PEP) of different papillary types were analyzed.Results:A total of 912 patients were enrolled in this study, and 86.95% (793/912) duodenal papilla conformed to one type of the classification, of which 77.18% (612/793) were regular type (type Ⅰ), 10.21% (81/793) small nipple type (type Ⅱ), 8.58% (68/793) protruding or drooping type (type Ⅲ), and 4.04% (32/793) wrinkled or ridged type (type Ⅳ). The success rates of SBC in four types of duodenal papilla were 98.86% (605/612), 90.12% (73/81), 88.24% (60/68) and 96.60% (28/32) respectively. The success rate of SBC of type Ⅰ was higher than those of type Ⅱ-Ⅳ (all P<0.008) and there was no significant difference between those of type Ⅱ and type Ⅲ ( P> 0.008). The pre-incision rate in the four types of duodenal papilla were 7.84% (48/612), 32.10% (26/81), 50.00% (34/68) and 25.00% (8/32) respectively. The pre-incision rate of type Ⅰ was lower than those of type Ⅱ-Ⅳ (all P<0.008), and there was no significant difference between those of type Ⅱ and type Ⅲ ( P>0.008). The difficult intubation rate in the four types of duodenal papilla were 12.42% (76/612), 39.51% (32/81), 58.82% (40/68) and 28.12% (9/32) respectively .The difficult intubation rate of type Ⅰ was lower than those of type Ⅱ and type Ⅲ (both P<0.001), and that of type Ⅲ was higher than that of type Ⅳ ( P=0.004) . There was no significant difference between those of type Ⅰ and type Ⅳ or type Ⅱ and type Ⅲ (both P>0.008). The incidences of PEP of the four types were 2.61% (16/612), 12.35% (10/81), 5.88% (4/68) and 6.25% (2/32) respectively. The incidences of PEP of type Ⅱ was higher than that of type Ⅰ ( P<0.001) , and there was no significant difference between those of type Ⅰ and type Ⅳ or type Ⅱ and type Ⅲ (both P>0.008). Conclusion:SBC is affected by duodenal papilla morphology, easiest for type Ⅰ and hard for type Ⅱ and Ⅲ. Attention should be paid to risk of PEP in SBC of type Ⅱ.

Objective To investigate the regulating mechanism of Notch1 signaling pathway on the invasion and migration ofglioma initiating cells (GICs).Methods (1) Box-plotting was conducted to analyze the mRNA expression of Notch1 in normal brain tissue and glioblastoma tissue using Bredel Brain,Sun Brain and TCGA databases;Kaplan-Meier survival analysis was conducted to analyze the association between the prognosis of glioma patients with the expression of Hes1 in TCGA database;Heatmap was conducted to analyze the expression of Notch1 and CXCR4 in GICs and common cell line in GEO database.(2) Magnetic activated cell sorting was adopted to establish cell lines of U87 GICs and U251 GICs;immunofluorescence staining was used to detect expression of CXCR4 and Notch1.After the cell lines of U87 GICs and U251 GICs were divided into DMSO,shNC,MK0752 and shNotchl groups,the shNotch1 and shNC groups were transfected respectively with recombinant lentivirus of Notch1-shRNA and its control sequence while the MK0752 and DMSO groups were added respectively with MK-0752 of 80 nmol/mL and the same amount of DMSO.The protein expression of Notch1,CXCR4 and p-mTOR was detected by Westem blotting in the 4 groups.The capabilities of invasion and migration of the GICs were detected by Transwell assay in the shNotch1 and shNC groups.Results (1) The box-plotting showed the mRNA expression of Notch 1 in the glioblastoma tissue was significantly higher than in the normal brain tissue (P＜0.05).The Kaplan-Meier survival analysis showed that the life span ofglioma patients with high expression of Hes1 was significantly shorter than that of those with low expression of Hes1 (P＜0.05).Heatmaps showed that the expression levels of Notch1 and CXCR4 in GICs were higher than in the common cell line.(2) The immunofluorescence staining showed that Notch1 and CXCR4 were highly expressed and colocalized in cell lines of U87 GICs and U251 GICs.The Western blotting showed that the protein expression of Notch1,CXCR4 and p-mTOR in the cell lines of U87GICs and U251 GICs in the MK0752 and shNotch1 groups was lower than that in the DMSO and shNC groups.Transwell assay showed that the penetrating-membrane cells per visual field in the shNotch1group were significantly fewer than those in the shNC group (P＜0.05).Conclusion Notch1 signaling pathway can promote invasion and migration of GICs through regulating CXCR4 expression.

Objective: To study the specific killing effect of CD4 membrane protein targeted chimeric antigen receptor modified T (CAR-T) cell. Methods: The second generation CD4 targeted chimeric antigen receptor containing 4-1BB costimulation domain was insert into lentiviral vector through recombinant DNA technology. Lentivirus was prepared and packaged by 293T cells with four plasmids. Beads activated T cells were transduced with lentivirus and the transduction efficiency was checked with Protein L and flow cytometry. T cell subsets and IFN-γ concentrations were detected with probe-tagged antibody and cytometric bead assay. Results: ①The transduction efficiency of activated T cells with prepared lentivirus were 50.0%-70.0%. A subset of CD8⁺ T cell acquired dim expression of CD4 membrane protein after activation. CD4⁺T cell and CD8⁺CD4^dim T cell were gradually killed by CD4 targeted CAR-T post lentivirus transduction. ②The kill efficacy of CD4 targeted CAR-T cell and control T cell toward KARPAS 299 T cell at an E∶T ratio of 8∶1 for 24 h was (96.9±2.1)% and (11.2±3.1)%, CAR-T cell has a higher killing efficacy than control T cell (t=7.137, P=0.028). The IFN-γ concentrations in culture supernatant of CAR-T cell with K562-CD4 cell, CAR-T cell with K562 cell and CAR-T cell alone were (15 648±2 168), (1 978±354) and (1 785±268) pg/ml, CAR-T cell cocultured with K562-CD4 cell produced more IFN-γ than the other two controls (P<0.01). Conclusions CD4 targeted CAR-T has an immunophenotype of CD8⁺CD4^-T cell. CD4 targeted CAR-T cell has killing efficacy toward normal CD4⁺T cell and CD4⁺T lymphoma cell. CD4 targeted CAR-T cell also has a killing efficacy toward CD4^dim target cell.

With the market-oriented application of new technologies and the development of various subjects in hospitals, more and more point-of-care test(POCT) devices appear in medical institutions, as an important method for rapid clinical acquisition of test results. As introduced by the authors, the hospital since 2017, established based on the hospital′s total quality management system, the POCT management committee, and built the hospital′s POCT quality management system. Thanks to its efforts, the hospital has fully standardized the POCT management from equipment procurement, test, maintenance, personnel training and examination, thus further ensuring medical quality.

During coronavirus disease 2019 epidemic, the treatment of severe trauma has been impacted. The Consensus on emergency surgery and infection prevention and control for severe trauma patients with 2019 novel corona virus pneumonia was published online on February 12, 2020, providing a strong guidance for the emergency treatment of severe trauma and the self-protection of medical staffs in the early stage of the epidemic. With the Joint Prevention and Control Mechanism of the State Council renaming "novel coronavirus pneumonia" to "novel coronavirus infection" and the infection being managed with measures against class B infectious diseases since January 8, 2023, the consensus published in 2020 is no longer applicable to the emergency treatment of severe trauma in the new stage of epidemic prevention and control. In this context, led by the Chinese Traumatology Association, Chinese Trauma Surgeon Association, Trauma Medicine Branch of Chinese International Exchange and Promotive Association for Medical and Health Care, and Editorial Board of Chinese Journal of Traumatology, the Chinese expert consensus on emergency surgery for severe trauma and infection prevention during coronavirus disease 2019 epidemic ( version 2023) is formulated to ensure the effectiveness and safety in the treatment of severe trauma in the new stage. Based on the policy of the Joint Prevention and Control Mechanism of the State Council and by using evidence-based medical evidence as well as Delphi expert consultation and voting, 16 recommendations are put forward from the four aspects of the related definitions, infection prevention, preoperative assessment and preparation, emergency operation and postoperative management, hoping to provide a reference for severe trauma care in the new stage of the epidemic prevention and control.