Establishment of a mouse model of acute systemic cold injury induced by hypothermia / 中华急诊医学杂志

Objective:To establish an animal model of acute systemic cold injury in mice.Methods:There were 98 C57BL/6 mice, half male and half female, with body weight of 22-27 g and age of 10 weeks. The mice were randomly divided into 7 groups ( n=14) according to the changes of anal temperature in cold environment, namely, group A (38.5 ± 1) ℃, group B (35 ± 1) ℃, group C (30 ± 1) ℃, group D (25 ± 1) ℃, group E (20 ± 1) ℃, group F (15 ± 1) ℃, and group G (10 ± 1) ℃, among which, group A was the blank control group, and the rest groups were the experimental group. The mice in the blank control group were placed in the normal environment (20 ± 5) ℃, and the mice in the experimental group were placed in the low temperature artificial climate box at - 20℃. The anal temperature of the mice was measured intermittently (as the core temperature), and the time required for the core temperature of the mice to drop to groups B, C, D, E, F and G was recorded. The righting reflex was used to evaluate the consciousness state, the action ability and the general state of each organ of mice were observed, and the blood routine and HE staining of each organ were detected. Results:The lower the core temperature of the experimental group, the longer the time required. The consciousness state, action ability, general state of organs, blood routine, and HE staining of organs in groups B, C, and D were basically the same as those in group A, and there was no acute systemic cold injury. Therefore, the blood routine, general observation of organs, and HE staining of organs in groups B, C, and D were no longer displayed compared with those in group A. Compared with group A, mice in group E began to suffer from disturbance of consciousness and action ability. With the decrease of core body temperature, the damage was aggravated, and mice in group G died. Compared with group A, the indices of blood routine test (WBC, RBC, HGB, PLT) of mice in group E began to decrease, and the univariate variance calculation showed that only WBC changes had statistical significance ( P<0.05). Compared with groups A and E, the indices of blood routine test (WBC, RBC, HGB, PLT) of mice in group F were further reduced, and the changes of each index in univariate variance calculation were statistically significant ( P<0.05). The general observation results showed that compared with group A, the lung, liver and spleen surfaces of mice in group E began to darken, and compared with groups A and E, the lung, liver, spleen, kidney and heart of mice in group F were further deepened and darkened, with irregular edges. HE staining results of various organs showed that compared with group A, the mice in group E began to have partial alveolar structure destruction and a small amount of inflammatory cell infiltration, the central vein of the liver was slightly congested, and the red and white pulp of the spleen were indistinct. Compared with groups A and E, the pathological structure damage of the lung, liver, spleen, kidney, heart and brain tissues of the mice in group F was further aggravated. Conclusions:Detection of consciousness state, action ability, general state of organs, blood routine and HE staining indices of organs in mice under low temperature can simulate the progress of clinical acute cold injury, and the animal model of acute systemic cold injury was successfully prepared.

An intelligent new method for batch weighing of blood components / 中国输血杂志

【Objective】 To establish a method for batch weighing blood components intelligently and accurately. 【Methods】 Virus inactivated plasma was used as weighing samples. The traditional electric scale and cryoprecipitation preparation instrument were used to weigh plasma with five specifications. The consistency of the weighing results by the two methods was statistically analyzed. Two groups of virus inactivated plasma with basically the same number of bags were randomly selected, and weighted by the same group of staff using these two equipment, the time for weighing and the time from weighing to registering were calculated statistically. 【Results】 227 bags of plasma were weighed in batches by the two methods, and statistical analysis showed no significant difference in standardized blood volume. There was no significant difference in plasma weighing time (71/68)by the same staff, weighing basically the same number of bags. However, the time difference from weighing to registering was statistically significant, as the new method was 2.65 s shorter than the traditional one (P<0.05). 【Conclusion】 The new method is accurate for batch weighing, and can realize the informatization and intellectualization of blood weighting. The time consumed from weighing to registration was improved by the new method, but not in time for weighing.

Study on the production efficiency of platelet components in 24 prefecture-level blood stations in China / 中国输血杂志

【Objective】 To learn the production efficient of platelet components among prefecture-level blood stations in China, to provide supporting data for those blood stations to optimize the production mode of platelet components and continuously improve production efficiency and supply capacity. 【Methods】 The data from 2017 to 2020 was obtained from 24 prefecture-level blood stations who were the members of the practice comparison network for blood institutes in China. The collection units of apheresis platelets, the number of dual-collections of apheresis platelets and plasma, the average apheresis units of one platelet apheresis procedure, the discarded rate of apheresis platelets, the amount of expired apheresis platelets and the amount of apheresis platelets issued were collected. For concentrated platelets, the prepared amount of platelet concentrates and the amount of expired platelet concentrates were collected; both the quantity of qualified and issued concentrated platelets were submitted for statistical analysis.The total output and efficiency of platelet components were calculated based on the collected data. 【Results】 The average annual growth rate of apheresis platelets collection in 24 prefecture-level blood stations was 12.23%, accounting for 99.80% of the total platelet output; the average collection unit of one platelets apheresis procedure was 1.75; from 2019 to 2020, only 5 blood stations performed dual-collection of platelet and plasma during one apheresis procedure; the discarded rate of apheresis platelets was 0.28%, of which 0.007% was due to expiration. A total of 1 621.2 therapeutic units of concentrated platelets were prepared, and 13.03% of them was discarded due to the expiration. The production efficiency of platelet components was 97.56%, of which the production efficiency of apheresis platelets was 97.61% and the production efficiency of concentrated platelets was 74.43%. 【Conclusion】 There are large regional differences in the supply capacity of platelet components in prefecture-level blood stations. Apheresis platelets are the main resource of platelet components product, and the collection capacity is increasing over the years with the characteristics of high production efficiency and low expiration scrapping rate. However, the preparation of concentrated platelets are still limited with relatively low production and high expiration discarded rate.

Study on proteomic changes of lung blast injury in mice / 中华急诊医学杂志

Objective:To establish the lung blast injury model in mice, detect the proteomic changes of lung in mice at different time points, and explore the mechanism of lung blast injury.Methods:A total of 60 healthy male C57BL/6 mice were randomly (random number) divided into the control group, 12-h group after thorax blast, 24-h group, 48-h group, 72-h group and 1-week group ( n=10 each group). Experiments were carried out in the animal laboratory of the General Hospital of the Northern Theater Command. The model of lung blast injury in mice was established by using a self-developed precision blast device, and the lung tissue injury situation was evaluated by gross observation and HE staining. The proteins in mouse lung tissue were quantitatively analyzed based on LC-MS/MS proteomic technology, and the differentially expressed proteins were screened. On this basis, bioinformatics tool was used to analyze proteomic changes. Results:After lung blast injury, scattered bleeding spots could be observed on the surface of lung tissue of mice, and the bleeding points were gradually increased with time, showing a patchy distribution, and the symptoms were the most severe at 24 h. The results of HE staining showed that the normal tissue structure of alveoli disappeared at 12 and 24 h under light microscopy with diffuse bleeding in the alveolar cavity, infiltration of a large number of inflammatory cells, increased interstitial exudate, thickened alveolar wall, and collapsed and merged alveolar cavity. A total of 6 861 proteins were identified by LC-MS/MS in lung tissue samples of mice after thorax blast, and 608 differentially expressed proteins were quantified, of which 227, 140, 202, 258 and 71 differential proteins were at 12 h, 24 h, 48 h, 72 h, and 1 week, respectively. According to GO analysis, 130 biological process subtypes including cell adhesion, extracellular matrix tissue and collagen fibril tissue were obtained. Besides, 66 cellular component involving extracellular exosomes, extracellular matrix and cytoplasm were obtained. And 43 molecular functional subclasses such as extracellular matrix structure composition, actin binding and antioxidant activity were obtained. KEGG analysis yielded 24 pathways including ECM-receptor interactions, focal adhesions and PI3K-Akt signaling pathway across the endothelium.Conclusions:Differentially expressed protein combinations are also different at different time points in the early stage after lung blast in mice, and the injury mechanism is complicated. The lung blast injury is the most serious at 12-24 h after blast and produces significant inflammatory response.

Establishment and study of a new mouse brain blast injury model / 中华急诊医学杂志

Objective To develop a new type of blast injury simulator to establish a mouse model of brain blast injury and study its damage mechanism. Methods Thirty healthy Kunming mice were randomly(random number) divided into the normal control group and brain blast injury model (TBI) group. A mouse model of traumatic brain injury was prepared by a self-developed explosive injury simulator. Morris water maze, Evans blue experiment and HE staining were used to observe the effects of shockwave exposure on spatial memory, blood-brain barrier, and pathological changes of brain tissues. T test was used for statistical analysis. Western blot method was used for detecting expression of brain injury markers Tau, S100β, Choline, inflammatory factors IL-1β, IL-4, IL-6, IL-10, NF-κB, apoptosis factors Bcl-2, Bax, Caspase3, and oxide protein stress-related factors IREα, MDA5, COX2 SOD1, and SOD2. Results Compared with the normal control group, (11.2±2.1) s, the time of searching platform in the TBI group was (54.6±8.4) s, was significantly longer (t=-19.330, P<0.05), and the EB exudation in the TBI group was 3.22 times (t=-13.903, P<0.05). Pathological staining revealed neuronal damage in the hippocampus, and TBI induced brain injury markers Tau(0.26±0.03 vs 0.46±0.04,t=-9.788, P<0.05), S100β(0.54±0.03 vs 0.74±0.02,t=-12.433, P<0.05) and Choline(0.54±0.05 vs 0.80±0.04, t=-7.970, P<0.05), inflammatory cytokines IL-1β(0.22±0.04 vs 0.31±0.05,t=-3.431, P<0.05), IL-4(0.65±0.02 vs 0.97±0.03, t=-18.927, P<0.05), IL-6(0.88±0.05 vs 1.07±0.08, t=-9.488, P<0.05) and NF-κB(0.80±0.06 vs 1.03±0.07,t=-4.507, P<0.05), and pro-apoptotic cytokines Bax(0.66±0.04 vs 0.78±0.04, t=-13.007, P<0.05) and Caspase3(0.44±0.03 vs 0.60±0.05, t=-4.472, P<0.05), oxidative stress-related factor pro IREα(0.72±0.06 vs 1.07±0.04, t=-9.665, P<0.05), MDA5(0.47±0.02 vs 0.77±0.02, t=-23.678, P<0.05) and expression of COX2(0.70±0.07 vs 0.86±0.02, t=-6.421, P<0.05), inhibition of inflammation inhibitory factor IL-10(1.14±0.06 vs 0.74±0.07, t=13.729, P<0.05), inhibition of apoptosis factors Bcl-2(0.72±0.05 vs 0.46±0.02, t=11.491, P<0.05) and inhibition of oxidative stress factors SOD1(1.17±0.05 vs 0.99±0.01, t=7.731, P<0.05) and SOD2(0.81±0.05 vs 0.61±0.04, t=10.257, P<0.05) expression. Conclusions The brain injury induced by blast exposure can induce spatial learning and memory loss, blood brain barrier disruption, neuronal damage hippocampus in mice, and promote the expression of brain injury markers, induce inflammation, oxidative stress and apoptosis. The self-developed explosive shock simulator successfully establishes a mouse brain blast injury model.