Objective To analyze the clinical value of plasma insulin-like factor 6(INSL6)in predicting the risk of acute aortic syndromes(AAS)and adverse outcomes.Methods A retrospective case-control trial was conducted on 194 AAS patients admitted to Department of Cardiovascular Diseases of Second Affiliated Hospital of Army Medical University between April 2021 and June 2023.Another 194 sex-,age-and BMI-matched individuals without aortic diseases were recruited from the health examination center between December 2021 and January 2024.Their plasma INSL6 level was measured with ELISA,and the general clinical data and results of some laboratory tests were collected and compared between the 2 groups.Spearman correlation analysis was used to assess the relationship between plasma INSL6 and other variables,receiver operating characteristic(ROC)curve was plotted to evaluate the diagnostic efficacy of INSL6 for AAS occurrence,multivariate conditional logistic regression model was utilized to analyze the association between plasma INSL6 and AAS onset,and Kaplan-Meier survival curve and multivariate Cox proportional hazards regression model was applied to analyze the relationship between acute-phase plasma INSL6 level and adverse prognosis in AAS patients.Results The plasma INSL6 level was significantly higher in AAS patients at acute phase than the control individuals[704.40(481.32～1 152.62)vs 141.24(107.60～163.72)pg/mL,P<0.001],but no statistical difference was observed in the level among the patients with different AAS subtypes(aortic dissection,intramural hematoma,and penetrating aortic ulcer).Spearman correlation analysis showed that the plasma INSL6 level was positively correlated with platelet count(r=0.325,P<0.001)and hemoglobin concentration(r=0.186,P=0.009),and negatively with IL-6(r=-0.182,P=0.011),INF-γ(r=-0.283,P<0.001),and D-dimer levels(r=-0.195,P=0.006).Multivariate conditional logistic regression analysis revealed that plasma INSL6 level was independently associated with the occurrence of AAS(OR=28.634,95%CI:7.267～112.820,P<0.001).ROC curve analysis further confirmed that the optimal cutoff value of plasma INSL6 in predicting AAS was 259.425 pg/mL,with a sensitivity of 95.9%and a specificity of 98.5%at this threshold.Kaplan-Meier curve analysis demonstrated that AAS patients with low INSL6 level had significantly lower cumulative survival rates(P=0.020)and event-free survival rates(P=0.004)than those with high INSL6 level(P<0.05).Multivariate COX regression analysis revealed that,after adjusting for sex,age,systolic blood pressure,ST classification,and surgical treatment,acute-phase INSL6 level was independently associated with all-cause mortality(HR:0.999,95%CI:0.999～1.000,P=0.023),AAS-related mortality(HR:0.999,95%CI:0.998～1.000,P=0.012),and composite endpoint events(HR:0.999,95%CI:0.999～1.000,P=0.026)in AAS patients during follow-up.Conclusion Plasma INSL6 level is closely associated with the occurrence and adverse prognosis of AAS,and the indicator is expected to be an effective biomarker for diagnosing AAS and predicting its prognosis.

Objective To identify genetic regulators of ionizing radiation(IR)sensitivity through clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)genome-wide screening.Methods A specialized single guide RNA(sgRNA)library was developed targeting 987 stress-response regulatory genes identified through Kyoto Encyclopedia of Genes and Genomes(KEGG),Reactome and gene ontology(GO)databases,followed by construction of sgRNA plasmids and packaging into a lentiviral library.Using colon adenocarcinoma Caco-2 cells as a model system,the Cas9-stable transgenic line was established via lentiviral transduction and antibiotic selection.Library-transduced cells underwent puromycin selection,followed by γ-irradiation(dose optimized via preliminary experiments).Post-irradiation phenotypic selection was conducted after 14 days,with subsequent next-generation sequencing of surviving cell populations to identify enriched/depleted sgRNAs.Candidate genes were functionally validated through orthogonal assays:cell counting kit-8(CCK-8)proliferation analysis,clonogenic survival assays and flow cytometric quantification of reactive oxygen species(ROS)and apoptotic markers.Results The optimized screening platform identified 281 radiation response genes(165 radiosensitive and 116 radioprotective candidates).Functional validation revealed Bcl2-associated athanogene 3(BAG3)knockdown significantly enhanced radioresistance.Proliferation was increased 1.2-1.5 fold,clonogenic survival improved 1.5-2.0 fold,and ROS was reduced by 13％-25％coupled with 32％-73％apoptosis attenuation compared to control groups.Conclusion The integrated CRISPR/Cas9 screening platform effectively identifies novel radiation response regulators,as evidenced by the discovery of BAG3 as a critical radiosensitivity determinant.This high-throughput functional genomics approach provides a robust methodology for systematically mapping molecular determinants of cellular radiation response,with potential applications in both mechanistic studies and therapeutic target discovery.

Objective To investigate the effect of erythroid differentiation associated gene(EDAG)on hematopoietic stem/progenitor cells(HSPCs)under chronic inflammation.Methods In this study,EDAG-/-and wild type(WT)mice were divided into the experiment group and control group.An infectious chronic inflammation model was established via multiple intraperitoneal injections of Listeria monocytogenes(LM),while a sterile chronic inflammation model was generated via multiple intraperitoneal injections of polyinosinic-polycytidylic acid[Poly(I∶C)].The effect of EDAG on HSPCs was explored under chronic inflammation conditions.Results In the LM repeated infection model,EDAG deletion led to a decrease in HSPCs and long-term hematopoietic stem cells(LT-HSCs)in mice as well as a significant bias towards myeloid differentiation in peripheral blood.Similarly,EDAG knockout also resulted in reduced numbers of HSPCs and decreased colony-forming ability in aseptic chronic inflammation models.Conclusion EDAG deficiency accelerates HSPC depletion in young mice under chronic inflammation,indicating strong protection of EDAG against HSPC damage induced by chronic inflammation.

Objective:To investigate the difference in the radiation sensitivity of hematopoietic stem and progenitor cells (HSPCs) derived from fetal liver and bone marrow.Methods:HSPCs from fetal liver of 14.5 d embryo or bone marrow of 8 week-old mice were isolated to receive a single dose of 5 or 10 Gy irradiation in vitro using a 60Co irradiator. Twelve hours later, the cell apoptosis, mitochondrial reactive oxygen species (ROS) level, colony formation ability and DNA damage in HSPCs were detected. Freshly isolated HSPCs were injected into lethally irradiated CD45.1 + C57BL/6J mice (4.5 Gy+ 5 Gy with an interval of 30 min) Chimerism rate, lineage constitution, and cell cycle were analyzed 12 weeks after transplantation. Results:Compared with bone marrow HSPCs after irradiation, the percentage of apoptosis in fetal liver HSPCs was significantly higher ( t=16.21, 12.27, P<0.05), the level of ROS was dramatically elevated ( t=68.72, 18.89, P<0.05). At 10 Gy, fetal liver HSPCs could not form colonies at all ( t=12.41, 15.67, 9.46, P<0.05). γ-H2AX immunofluorescence staining showed that the DNA damage of fetal liver HSPCs was more severe after irradiation, and the number of Foci formed was significantly higher than that of bone marrow HSPCs ( t=2.27, 2.03, P< 0.05), which indicated that fetal liver HSPCs were more sensitive to radiation. The chimerism rate of transplanted fetal liver HSPCs was lower than that of bone marrow cells ( t=5.84, P<0.05) with a higher proportion of myeloid lineage, suggesting that fetal liver HSPCs had lower in vivo reconstitution capacity than bone marrow HSPCs and were more prone to myeloid differentiation. The cell cycle of bone marrow HSPCs from transplanted chimeric mice was examined, and the proportion of S-phase was significantly higher in the fetal liver group than that in the bone marrow group ( t=2.89, P<0.05). Mitochondrial stress results showed that fetal liver HSPCs had higher basal respiratory capacity ( t=39.19, P<0.05), proton leakage ( t=6.64, P<0.05), ATP production ( t=9.33, P<0.05), and coupling efficiency ( t=7.10, P<0.05) than bone marrow c-Kit + cells, while respiratory reserve capacity ( t=5.53, P< 0.05) was lower than that of bone marrow c-Kit + cells. Conclusions:HSPCs derived from fetal liver display higher radiosensitivty compared with bone marrow HSPCs, laying the foundation for an in-depth illustration of the effects of radiation on hematopoietic stem cells at different developmental stages.

Objective:To study the effect of different doses of 60Co γ-ray ionizing radiation on mitochondrial function in mouse hematopoietic stem and progenitor cells (HSPCs). Methods:C57BL/6 mice were divided into control group, 1 Gy irradiation group and 4.5 Gy irradiation group. The mitochondrial functions were detected at 12 h and 24 h after irradiation, including ROS level, membrane potential, mitochondrial structure, and mitochondrial stress. Bone marrow c-Kit + cells received a single 15 Gy irradiation in vitro, after 24 h, mitochondrial function was detected. Results:It was found that mice leukocytes ( t=12.41, 18.31, 16.48, 14.16, 19.08, 20.25, P<0.05), red blood cells ( t=4.81, 6.62, P<0.05) and platelets ( t=4.33, 6.68, P<0.05) were significantly reduced. The numbers of bone marrow colony formation unit ( t=16.27, 55.66, 17.06, 43.75, P<0.05), and HSPCs ( t=5.16, 11.55, P<0.05) were decreased dose-dependently post-irradiation. Under 1 Gy irradiation, the mitochondrial function and mitochondrial basal metabolic index of HSPCs ( t= 7.36, 3.68, 4.58, 3.15, 3.15, P<0.05) were enhanced at 24 h post-irradiation. Under 4.5 Gy irradiation, mitochondrial number, mitochondrial membrane potential ( t=12.29, 10.46, P<0.05), maximal respiration and spare respiratory capacity were decreased ( t=7.81, 5.78, 6.70, 5.83, P<0.05), ROS level was increased ( t=4.63, 4.12, P<0.05). The basal respiration and oxidative phosphorylated ATP production were reduced at 12 h after irradiation ( t=8.48, 3.80, P<0.05); and the proton leakage was increased ( t=6.57, P<0.05) and coupling efficiency was reduced ( t=11.43, P<0.05) at 24 h after irradiation. In cultured c-Kit + cells, the level of ROS ( t=11.30, P<0.05) and the maximum respiration and spare respiratory capacity were increased ( t=4.25, 3.44, P<0.05) while the mitochondrial membrane potential was decreased ( t=34.92, P<0.05) significantly. Conclusions:A method for systematically assessing mitochondrial function in HSPCs was established, and the effect of ionizing radiation on mitochondrial function of HSPCs was clarified, laying a foundation for further revealing the mechanism of ionizing radiation-induced mitochondrial damage in HSPCs.

To report a 4-year-old boy with severed right index finger amputations in 2 segments. There were severe contusion on the 2 amputated sections of finger. According to the prevention and control requirement for the novel coronavirus disease(COVID-19), the patient was firstly checked to exclude the COVID-19. Then the replantation surgery was successfully carried out under the strict protective measures. The replanted index finger survived well at 2 weeks after surgery.

Objective To summarize the clinical experience of extrocorporeal membrane oxygena-tion(ECMO) in the treatment of pediatric acute respiratory distress syndrome (ARDS). Methods A retro-spective analysis of children with ARDS who were hospitalized for different causes and received the treatment of ECMO from October 2012 to November 2017 was performed. The clinical conditions and prognostic fac-tors in the course of their disease were compared. Results In 12 cases of ARDS,9 cases (75％ ) had severe pneumonia,2 cases (16. 67％ ) had lung tumor resection and 1 case ( 8. 33％ ) had bronchial foreign body. Seven cases (58. 3％ ) chose VA-ECMO,5 (41. 7％ ) cases chose VV-ECMO. The average duration of ECMO was (225. 03 ± 214. 75) h. With the positive treatment of ECMO,heart rate,mixed venous oxygen saturation and central venous pressure all improved significantly(P < 0. 05),and there was no obvious abnor-mal changes in MAP and lactic acid(P > 0. 05). Arterial oxygen partial pressure,arterial carbon dioxide par-tial pressure,oxygenation index and P/ F were significantly improved after the ECMO support(P < 0. 05). Ppeak,Paw and PEEP after evacuation of ECMO were significantly lower than those before treatment (P <0. 05). Ten cases (83. 33％ ) were successfully removed,8 cases (66. 67％ ) were saved, and 4 cases (33. 33％ ) died. During the ECMO treatment,9 cases (75％ ) had complications,including 8 cases of bleed-ing at the intubation site,3 cases of gastrointestinal hemorrhage,2 cases of hemolysis,1 case of infection,2 cases of acute kidney injury,2 cases of neurological symptoms,1 case of multiple organ dysfunction syn-drome. Conclusion Pediatric ARDS is critical and the mortality rate is high. ECMO should be used as soon as possible when the lung is potentially regained and other treatments are ineffective,so that the lung could be fully rescued to gain time and opportunity for clinical treatment.

Objective: To investigate the application and outcome of pediatric extracorporeal membrane oxygenation (ECMO) in a single center. Methods: The clinical data of 52 pediatric patients with cardiopulmonary failure received ECMO support in Bayi Children's Hospital Affiliated to General Hospital of Beijing Military Command of PLA were collected from January 2012 to October 2016. All patients were divided into two stages by time. January 2012 to December 2014 was stage one. January 2015 to October 2016 was stage two. A retrospective analysis was done for these patients between two stages. In addition, all clinical data were compared with the data of extracorporeal life support organization (ELSO). The constituent ratio differences in different groups were tested by chi square test. Results: In 52 cases, there were 40 boys and 12 girls, aging from 1 day to 7 years, weighing from 2 to 20 kg. There were 35 cases who successfully weaned from ECMO (67%), and 25 cases were able to be discharged alive (48%). In stage one, there were 24 ECMO cases, 18 boys and 6 girls. There were 15 cases successfully weaned from ECMO (63%). Nine patients survived until discharge (38%). Complications were found in 15 cases during ECMO support (63%). In stage two, there were 28 ECMO cases, 22 were boys and 6 were girls. There were 20 cases successfully weaned from ECMO (71%). Sixteen patients survived until discharge (57%). Complications were found in 12 cases during ECMO support (43%). There was no significant difference in survival rates between two stages. However, the neonatal survival rate was higher in stage two than in stage one (71% (12/28) vs. 31% (5/24), χ²=5.107, P=0.038). The proportion of respiratory support was higher in stage two than in stage one (50% (14/28) vs. 21% (5/24), χ²=4.741, P=0.029), while the proportion of extracorporeal cardiopulmonary resuscitation (ECPR) decreased significantly (21% (6/28) vs. 67% (16/24), χ²=10.835, P=0.001). Application of peritoneal dialysis treatment in stage two was higher (6 vs. 0 cases, χ²=8.097, P=0.025). Mortality of ECMO was still higher than that of ELSO (48% (25/52) vs. 62% (34 655/55 886), χ²=4.281, P<0.05). The constituent ratio of different types of support varied between ECMO and ELSO patients (χ²=19.562, P<0.001). Conclusions ECMO technology can provide effective support for severe cardiopulmonary failure in critically ill children. Due to the multidisciplinary nature of ECMO technology, the complexity and characteristics of pediatric patients, it takes long time to improve ECMO management and prognosis.

Objective: To retrospectively review 4 cases diagnosed with pediatric acute respiratory distress syndrome (ARDS) who were transported with veno-venous (V-V) extracorporeal membrane oxygenation (ECMO) from April 2016 to March 2017. Methods: Four patients were transported to Bayi Children's Hospital Afflicted to the PLA Army General Hospital, with V-V ECMO. Their vital signs, blood-gas analysis and chest X-ray before and after transportation were compared. The length of ECMO, pediatric intensive care unit (PICU) stay and hospitalization, and the prognosis were analyzed. Results: All the four cases were transported to our hospital successfully from distances between 1 000 km to 1 210 km. The 4 cases were 4 to 6 years old with the body weight of 19 to 35 kg, of whom 3 were boys and 1 was girl. The catheters were inserted in the right jugular vein and femoral vein. The vital signs and blood-gas analysis after transportation did not change significantly compared to baseline. The length of ECMO for the four patients were 48, 754, 157 and 438 hours. They stayed in the PICU for 10, 32, 14 and 19 days, respectively. At last, 2 of them were successfully discharged from hospital without any complications; however, the other 2 died of multiple organ failure. Conclusion Transporting ARDS patients with a satisfactory cardiac function under VV-ECMO by an experienced ECMO team is safe.

Objectives: To explore the expression regulation of type 1 and type 2 (Th1 and Th2) cytokines from serum of coal miners and the evaluation in surveillance of coal workers' pneumoconiosis, 630 coal miners were studied. Methods: A total of 90 male patients diagnosed as coal workers' pneumoconiosis (CWP) in a institute for occupational health and 19 male workers newly diagnosed as CWP patients was chosen as CWP group with simple random sampling method from a coal mine group from January 2013 to December in 2015. 180 male coal miners with abnormal but not diagnosed as CWP were selected as CWP suspected group with simple random sampling methods, meanwhile 180 male coal miners with normal chest X-ray photograph was as dust-exposed group by 1∶1 matched as age. And 161 healthy males accepted pre-employed examination were selected as control group, CWP suspected group, dust-exposed group and control group called as non-CWP group. According to screening test and diagnosis test, the basic information and occupational history of all subjects were collected, and cytokines including IL-1β, IL-8, IFN-γ, IL-6 and IL-10 of serum were detected. Receiver operator characteristic (ROC) curve was used to determine the optimal cutoff value of each cytokine. Area under curve (AUC), the validity and reliability were calculated and judged. Results: The average age of control group, dust-exposed group, CWP suspected group and CWP group were (27.4±5.0) , (43.4±10.7) , (48.2±6.2) , (64.7±7.0) years old, respectively. The median level of IL-1β, IL-8, IFN-γ and IL-6 in cases group (1 638.30, 2 099.49, 815.18,140.32 pg/ml) were higher than that of non-cases group (1 445.57, 1 402.26, 736.38, 95.73 pg/ml) (P<0.05) . The level of IL-8 (1 503.99 pg/ml) in CWP suspected group was higher than that of control group (1 295.67 pg/ml) and dust-exposed group (1 376.94 pg/ml) , but the level of IL-10 (654.08 pg/ml) was lower than that of control group (596.64 pg/ml) . The ratio of IFN-γ/IL-6 ranged from 5 to 8, and the ratio in CWP group (5.87) was lower than that of non-CWP group (7.61) . The IL-6 and IL-8 among the subjects of dust-exposed group in terms of the age distribution of among had reached statistical significance. According to ROC, the cutoff value of IL-1β, IL-6, IL-8, IL-10 and INF-γ reached 1 582.65, 116.53, 1 791.54, 581.08 and 792.69 pg/ml, respectively. The AUC was 0.668, 0.895, 0.859, 0.716 and 0.637, respectively. It was found that IL-6 and IL-8 could be used as biomarkers in detecting CWP, the sensitivity and specificity was 82.6% and 84.6%, 78.0% and 84.8%, respectively; Youden's index was 0.674 and 0.628 and the consistency rate was 84.3% and 83.7%, while Kappa value was 0.55 and 0.52. Conclusion There was Type 1 and type 2 cytokine dysregulation in CWP patients. IL-6 and IL-8 can be used as effective biomarkers to forecast lung injury before X-ray changes.