AIM: To investigate the relationship between vascular smooth muscle cell (VSMC) injury, organelle stress response and autophagic cell death (autophagy) and ferroptosis induced by the chemical hypoxia inducer cobalt chloride (CoCl2) through the bioinformatics analysis and in vitro cell experimentation. METHODS: The dataset GSE119226 of VSMC treated with cobalt chloride was acquired from the gene expression database (GEO). The R language was used to investigate the relationship between CoCl2 treatment and organelle stress response (Golgi stress, endoplasmic reticulum stress) and two forms of cell death (ferroptosis and autophagic cell death). With primary cultured rat VSMC (rVSMC) and CoCl2-induced anoxia model, the changes in cell viability were detected by CCK-8 method, and reactive oxygen species (ROS) levels were measured using DCFH-DA method. The expression levels of HIF-1α (a key molecule in hypoxia), Golgi stress markers GM130 and p115, endoplasmic reticulum stress markers GRP78 and CHOP, autophagy markers LC3-II / LC3-I and Beclin1, and ferroptosis markers GPx4 and xCT were detected by Western blot. The effect of inducing or inhibiting organelle stress and cell death on the CoCl2-induced cell damage was also observed. RESULTS: Differentially expressed genes analysis of GSE119226 dataset showed that CoCl2 treatment of VSMCs had significant effects on organelle function and stress response, autophagy and ferroptosis-related genes, in which endoplasmic reticulum stress, protein processing in endoplasmic reticulum, regulation of Golgi to plasma membrane protein transport, autophagy / autophagic cell death, and ferroptosis pathways were remarkably enriched. The results of in vitro experiment showed that compared with normal rVSMC, cell viability was significantly decreased after CoCl2 treatment, as well as HIF-1α protein expression and ROS levels in rVSMCs were increased. In rVSMC treated with Co-Cl2, the expression levels of Golgi structural proteins GM130 and p115 (reflecting the occurrence of Golgi stress) were decreased, while the markers GRP78 and CHOP (reflecting the occurrence of endoplasmic reticulum stress) were increased. At the same time, CoCl2 treatment also reduced the expression of autophagy markers LC3-II/LC3-I and Beclin1 (indicating the decrease levels of autophagy), while the expression of ferroptosis markers GPx4 and xCT were decreased (indicating the occurrence of ferroptosis). Compared with CoCl2 treatment group, induced Golgi stress, endoplasmic reticulum stress, or ferroptosis could further reduce cell viability, while inhibition of these processes could improve cell viability. On the other hand, increasing the level of autophagy can improve the cell viability. CONCLUSION: Hypoxia induced by cobalt chloride can lead to VSMC injury. Golgi stress, endoplasmic reticulum stress, ferroptosis, and the reduction of autophagy level play an important role in it. Inhibition of organelle stress response and ferroptosis, or increase of autophagy level can improve VSMC injury caused by cobalt chloride.

Objective:To discuss the effect of fluid resuscitation on the occurrence of ferroptosis in vascular tissue and the structure of vascular cells in the rats with blast injury complicated with hemorrhagic shock,and to clarify its mechanism.Methods:A total of 54 healthy adult SD rats were randomly divided into normal group,blast injury complicated with hemorrhagic shock(model)group,and the fluid resuscitation(treatment)group,and there were 18 rats in each group.Among them,10 rats were randomly selected to observe the surival status and another 8 rats were selected to detect the other indexes.The average survival time(ST),24 h and 72 h survival rates of the rats in various groups were observed;the blood pressure(BP),heart rate(HR),and respiratory rate(RR)of the rats in various groups were observed;the levels of serum creatinine(Scr),blood urea nitrogen(BUN),lactate(LAC),glucose(GLU),iron ions,glutathione(GSH),and malondialdehyde(MDA)and the activities of aspartate aminotransferase(AST),alanine aminotransferase(ALT)and lactate dehydrogenase(LDH)in serum of the rats in various groups were detected;Western blotting method was used to detect the expression levels of ferroptosis marker proteins glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),and heme oxygenase 1(HO-1)proteins in superior mesenteric artery tissue of the rats in various groups;the pathomorphology of the superior mesenteric artery of the rats in various groups was observed.Results:All the rats in normal group survived for 72 h,while the longest ST of the rats in model group did not exceed 9 h.Compared with model group,the ST and 24 h survival rate(SR)of the rats in treatment group were significantly increased(P＜0.05).Compared with normal group,the BP,HR,and RR of the rats in model group were significantly decreased(P＜0.01).Compared with model group,the BP,HR,and RR of the rats in treatment group were significantly increased after fluid resuscitation(P＜0.05).Compared with normal group,the activities of AST and ALT,and the levels of Scr and BUN in serum of the rats in model group were significantly increased(P＜0.01).Compared with model group,the serum levels of LAC and GLU of the rats in treatment group were significantly decreased(P＜0.01).Compared with normal group,the concentration of iron ion,GSH level,MDA level,LDH activity in serum of the rats in model group were significantly increased(P＜0.05);compared with model group,the concentration of iron ion and LDH activity in serum of the rats in treatment group was significantly decreased(P＜0.01).Compared with normal group,the expression levels of GPX4 and SLC7A11 in superior mesenteric artery tissue of the rats in model group were significantly decreased(P＜0.05);compared with model group,the expression levels of GPX4 and SLC7A11 in superior mesenteric artery tissue of the rats in treatment group were significantly increased(P＜0.05).Compared with normal group,the expression level of HO-1 protein in superior mesenteric artery tissue of the rats in model group was increased(P＜0.01);compared with model group,the expression level of HO-1 protein in superior mesenteric artery tissue of the rats in treatment group was increased(P＜0.01).The microscopic pathology results showed that the cell arrangement in the layers of the superior mesenteric artery tissue of the rats in model group was disordered,the swelling was significant and the thickness was increased;the pathological changes in superior mesenteric artery tissue of the rats in treatment group was alleviated.The ultramicroscopic pathology results showed that the endothelial cell structure of blood vessels of the rats in normal group was intact,and there was no swelling in the subendothelial matrix;the vascular endothelial cell membrane of the rats in model group was damaged,there were cytoplasmic dissolution and fragmentation,and the swelling of the subendothelial matrix was significant;the swelling of the vascular endothelial cells in treatment group was alleviated.Conclusion:Ferroptosis occurs in vascular tissue of the rats with blast injury complicated with hemorrhagic shock,and fluid resuscitation can alleviate the structural damage of the vascular cells by inhibiting the vascular tissue ferroptosis.

Objective To establish a model of combined injury of thermal radiation and blast wave of gas explosion in closed pipeline,and investigate the characteristics of important organ injury induced by burn-blast combined injury in rats.Methods A total of 72 male SD rats(aged 8～10 weeks,weighing 200～10 g)were randomly divided into control group and mild and severe injured groups.After the model rats were inflicted with blast wave and thermal radiation,their physical parameters were detected and measured.The respiratory function of the survival rats was tested.In 24 h later,arterial blood gas analysis,blood biochemical tests,and detection for serum inflammatory factors and lung injury related protein levels were performed,and the pathological changes in the lung tissue and trachea were observed.Results The peak range of explosive blast wave overpressure was 209～493 kPa,and the temperature was 152～258 ℃.The mortality rate was 8.3％in the mild group and 53.1％in the severe group.Compared with the control group,longer inspiratory time(Ti)and relaxation time(Tr)and larger tidal volume(TV)(P＜0.05),while lower respiratory frequency(f)were observed in the 2 injured groups within 6 h after injury(P＜0.05).At 24 h after injury,the values of partial pressure of carbon dioxide(PaCO2),residual base value(BE)and bicarbonate(HCO3-)were increased significantly(P＜0.05),while partial pressure of oxygen(PaO2)was decreased(P＜0.05).The serum levels of alanine transaminase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total bilirubin(TBI)and total bile acid(TBA)were notably elevated(P＜0.05).So were the levels of IL-1 β,TNF-α and IL-6(P＜0.01).Various severities of hyperemia foci and inflammation in the tracheal mucosa were observed in the injured rats.The incidence of lung injury was high,and the pathological manifestations were dot-shaped hemorrhage to diffuse hemorrhage of the whole lobe,rupture of alveolar septum,thickening of septum,and infiltration of red blood cells and inflammatory cells.The levels of serum lung injury-related proteins were significantly increased(P＜0.05).Conclusion A rat model which can highly simulate mild to severe burn-blast combined injury is constructed,which can meet the needs for rat model in study of explosive burn-blast compound injury.The main characteristics of the injury are lung injury,respiratory dysfunction,liver damage and secondary infection.

Objective:To investigate the effects of high-energy shock and vibration on cortex injury and peripheral blood immune cells in goats.Methods:Seventeen Boer goats without gender preference were selected. By using random number tables, the goats were divided into normal control group ( n=5) and shock and vibration injury group ( n=12). The goats in the normal control group were anatomized routinely and their brain was collected after being sacrificed without any other treatment. The goats in the high-energy shock and vibration model group were placed on a loading table (part of the BY10-100 instant shock and vibration simulation platform) in a restrained state, and made into a high-energy shock and vibration injury model induced by a vertical impact waveform generator. The intravenous blood samples were taken from the goats in the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury.Then, the goats were sacrificed and the following procedures were the same as the normal control group. At 24 hours after injury, the brain injury and the histopathological changes of the cerebral cortex in the normal control group and shock and vibration injury group were observed by gross pathological and anatomical examination and HE staining. The mRNA expression of zonula occludens 1 (ZO-1), tight junction protein 5 (Claudin-5), glial fibrillary acidic protein (GFAP), ionized calcium binding adaptor molecule 1 (IBA-1), interleukin (IL)-1β, IL-6 and cluster of differentiation antigen 177 (CD177) of the cerebral cortex in the normal control group and shock and vibration injury group were measured through fluorogenic quantitative polymerase chain reaction. The expression of ZO-1 and Claudin-5 proteins of the cerebral cortex in the normal control group and shock and vibration injury group were detected by Western blotting. Hematology analyzer and coagulation analyzer were used to detect white blood cell count, neutrocyte, lymphocyte, monocyte, prothrombin time 1 (PT-1), prothrombin time international normalized ratio (PT-INR), activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin activity (PTA) and fibrinogen (FIB) levels in goats of the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury, respectively. Results:At 24 hours after injury, no visible contusion or necrosis was found in goat brain tissue in the shock and vibration injury group; the cerebral micro-vessels presented with a local dilation, hyperemia, edema, aggregation of inflammatory cells, disruption of vessel walls and leakage of red blood cells. These changes were not observed in the normal control group. In the shock and vibration injury group, ZO-1 and Claudin-5 mRNA expressions in the cerebral cortex were 0.25±0.10 and 0.09(0.04, 0.44) respectively, which were significantly lower than those of the normal control group [1.00±0.15 and 0.99(0.80, 1.20)]; GFAP, IBA-1, IL-1β, IL-6 and CD177 mRNA expression levels were 4.40(3.88, 6.75), 2.60±1.07, 3.04±0.51, 2.71±0.45 and 2.93±0.62 respectively, which were significantly higher than those of the normal control group [1.00(0.78, 1.22), 1.00±0.37, 1.00±0.27, 1.00±0.57 and 1.00±0.35]; ZO-1 and Claudin-5 protein expression levels were 0.41±0.06 and 0.42±0.11 respectively, which were significantly lower than those of the normal control group (1.08±0.12 and 0.91±0.23) (all P<0.01). In the shock and vibration injury group, the levels of white blood count, neutrocyte, and lymphocyte in peripheral blood were (13.7±3.3)×10 9/L, (35.3±14.8)% and (57.2±15.1)% respectively before injury, (19.4±3.1)×10 9/L, (60.5±12.5)% and (33.6±14.2)% respectively at 3 hours after injury, and (20.6±3.6)×10 9/L, (63.6±13.0)% and (30.9±15.0)% respectively at 6 hours after injury. By contrast, the levels of white blood count and neutrocyte were significantly increased but the level of lymphocyte was significantly decreased at 3 and 6 hours after injury ( P<0.05 or 0.01); the levels of the above indicators showed no significant changes at 0 and 24 hours after injury (all P>0.05); the level of monocyte did not change significantly at all time points before and after injury (all P>0.05). The levels of PT-1, PT-INR, APTT, TT, PTA and FIB in the shock and vibration injury group did not change significantly at each time point before and after injury (all P>0.05). Conclusion:Cerebral cortex microvascular injury and disruption of blood-brain barrier can be initiated in the early stage of high-energy shock and vibration injury in goats, accompanied by the presence of central and peripheral inflammatory response.

Objective? To explore the effects of motivational interviewing rehabilitation transitional care based on action research on rehabilitation and functional recovery among stroke patients. Methods? From January 2017 to December 2017, we selected 80 stroke patients treated in Xinyang Central Hospital as subjects by convenience sampling. All the patients were divided into control group (n=40, routine discharge nursing for 12 months) and observation group (n=40, motivational interviewing rehabilitation transitional care based on action research for 12 months) with the random number table. The National Institutes of Health Stroke Scale (NIHSS), Fugl-Meyer Assessment (FMA), Activity of Daily Living (ADL) and Hamilton Depression Scale (HAMD) were used to evaluate the recovery of nerve function, motor function, ADL and depression of patients at discharge and 6 months after discharge, and we also compared the satisfaction with nursing between two groups. Results? At discharge, there were no statistical differences in the scores of NIHSS, FMA, ADL and HAMD of patients between two groups (P＞0.05). Six months after discharge, the scores of NIHSS, ADL and HAMD of patients in observation group were lower than those in control group;the score of FMA in observation group was higher than that in control group all with statistical differences (P＜0.01),and the satisfaction rate of the observation group was higher than that of control group with statistical difference (P＜0.01). Conclusions? The motivational interviewing rehabilitation transitional care based on action research can accelerate the process of rehabilitation of stroke patients, relieve depression, improve their ADL, quality of life and satisfaction with nursing.

Objective To develop an uncertainty study model of the enzymological reference method to systematically research the uncertainty influencing factors of 6 enzymological reference methods ALT,AST,LDH,AMY,GGT and CK for determining the key factors influencing the reference method and better guiding the establishment and operation of the enzymological reference methods.Methods The uncertainty of the six enzymological reference methods was evaluated by the theoretical evaluation com-bined with the experiment design according to series of standards including JJF1059-1999 Evaluation and Expression of Uncertainty in Measurement,JJF1135-2005 Evaluation of Uncertainty in Chemical Analytical Measurement,CANA-GL06 Guidance on Evalua-ting the Uncertainty in Chemical Analysis and CNAS-CL33 Guidance on the Application of Testing and Calibration Laboratories Competence Accreditation Criteria in the Field of Clinical Enzymology Reference Measurement.Results The established enzymo-logical uncertainty study model was C=ΔAΔt · 1ε · 1L ·V 1 +V 2 +V SV ·fT ·f pH ·fλ·fkit ·fresult .Conclusion The uncertainty e-S valuation model is set up and successfully applied in the uncertainty evaluation of the six enzymological reference methods.

Objective As a collaborator of Beijing Institute of Medical Device Testing for value assignment of state standard ma-terial candidate Cystatin-C ,we have used the internationally accepted reference material to assign value for state standard material candidate Cystatin-C ,and help Beijing Institute of Medical Device Testing get Cystatin-C national standard material certificate . Methods According to the target value and operational procedure of international reference material ERM-DA471 ,We have tested 6 dilutions of standard material candidate Cystatin-C on calibrated Hitachi 7180 immunoassay system .Results The results demon-strate good repeatability and commutability ,and have been accepted in calculating the final value for the candidate standard materi-al ,our data has assisted Beijing Institute of Medical Device Testing in passing the criteria and obtaining Cystatin-C national standard material certificate .Conclusion Compared to the data from all participating collaborators ,our results hit right on the target value , and no significant matrix effects have been observed .

Objective To approach the methods and effects of internal fixation for Hoffa fracture. Methods A total of 26 patients with 26 condylar Hoffa fractures ( medial condylar fractures in 13patients and lateral condylar fractures in 13) treated from August 1993 to February 2010 were retrospectively analyzed.According to the Letenneur classification,there were 16 patients with type Ⅰ fractures,four with type Ⅱ fractures and six with type Ⅲ fractures.Among them,two patients were with open fractures and 24 with closed fractures.Surgical approaches including screw fixation in 21 patients and lateral support plate fixation in five were selected based on the fracture types and affected sides. Results All patients were followed up for 12.5-48 months (average 18 months),which showed fracture healing in all the patients within 3-4 months (average 3.5 months).Two patients had slight shift together with knee joint pain,ie,one patient had ROM of knee for 95 °,and one failed the functional exercise because of pain and had ROM of knee for 60° during follow up.No complications including infection,delayed union or bone necrosis occurred.According to Letenneur' s functional assessment system,the postoperative outcomes were excellent and good in 24 condyles,fair in one and poor in one. Conclusions Surgical treatment for Hoffa fractures is safe and effective,but the key point is to choose correct screw fixation position and orientation according to the fracture types and fracture fragment size.

Objective To study the effect of nucha electroacupuncture (NEA) on 5-hydroxytryptamine (5-HT) in cerebrospinal fluid (CSF) and blood of acute cerebral infarction (ACI) patients. Methods 72 cases with first ACI were randomly divided into NEA group (n= 38) and control group (n=34). Their 5-HT levels in CSF and blood were determined before and 4 weeks after treatment. Results Before treatment, the 5-HT in the CSF was (0.67±0.13) μmol/L in the NEA group and (0.71±0.11) μmol/L in the control group (P>0.05), while that in the blood was (0.44±0.19) μmol/L in the NEA group and (0.41±0.10) μmol/L in the control group (P>0.05). After treatment, the 5-HT in the CSF was (1.12±0.32) μmol/L in the NEA group and (0.83±0.15) μmol/L in the control group (P<0.05), while that in the blood was (0.87± 0.14) μmol/L in the NEA group and (0.63±0.07) μmol/L in the control group (P<0.05). Conclusion NEA can increase the 5-HT in both CSF and blood after ACI, which may facilitate to reduce post-stroke depression.

Objective: To investigate the effects of complex prescription astragalus mongholicus injection(复方黄芪注射液) on the serum concentrations of neuron-specific enolase(NSE),myelin basic protein(MBP) and S100 protein B(S100B) in cases with acute severe craniocerebral injury.Methods: One hundred and ninety-six patients with acute severe craniocerebral injury were randomly divided into two groups.The treated group was treated with complex prescription astragalus mongholicus injection plus conventional treatments including dehydration,antibiotics,organ functional support,nerve nutrition,prevention of complication,etc;the control group was treated with conventional treatments alone.The concentrations of NSE,MBP and S100B in plasma at admission and at 4,7 and 10 days after treatment were determined;the Glasgow coma score(GCS) at admission and at 1 week and 2 weeks after hospitalization and the Glasgow outcome scale(GOS) after 3 months were compared to observe the long-term efficacy in the patients.Results: After treatment,the concentrations of serum NSE,MBP and S100B in the treatment group were all lower than those in the control group,the differences being significant(NSE(14.62?3.38)?g/L vs.(21.54?5.68) ?g/L,MBP(7.52?1.06) mg/L vs.(10.21?2.01) mg/L,S100B(0.90?0.28) ?g/L vs.(1.20?0.34) ?g/L,all P0.05);the GCSs of the patients at 1 week,2 weeks and GOS at 3 months after treatment in the complex prescription astragalus mongholicus injection group were significantly higher than those in the control group(GCS, 1 week(9.8?2.6)score vs.(7.2?2.1) score,2 weeks(10.6?3.0) score vs.(7.8?2.2) score;GOS,3 months after treatment(4.8?1.0) score vs.(3.6?0.8) score,all P