Hemostatic Effects of Bio-Zeolite Gauze and QuikClot Combat Gauze on Major Bleeding in Rabbits Acutely Exposed to High Altitude.

Objective: Hemostatic gauze application is an effective way to control major bleeding, which is the most common cause of death in trauma in both civilian and military settings. Coagulation derangement after acute exposure to high altitude might alter the effects of hemostatic gauzes. The present study aimed to observe the hemostatic effects of bio-zeolite gauze (BZG) and QuikClot Combat Gauze® (QCG) on major bleeding in rabbits acutely exposed to high altitude.Methods: Sixty rabbits were randomly and evenly divided into six groups. Animal models of simulated blast- and fragment-induced inguinal major bleeding were prepared in lower altitude and high-altitude areas, and BZG, QCG, and ordinary gauze without hemostatic material were used to control bleeding. The primary outcomes included immediate hemostasis rate, blood loss, and survival rate, while the secondary outcomes included hemodynamic parameters, laboratory examinations, and coagulation-relevant markers.Results: The overall effects of BZG and QCG were better than those of ordinary gauze, with a higher immediate hemostatic rate, less blood loss, and higher survival rate at 90 min after gauze application and higher red blood cell and platelet counts and lower creatinine level at 30 min after gauze application in lower altitude. The concentrations of coagulation factor XII and factor X in rabbits acutely exposed to high altitude were significantly lower than those in lower altitude. At high altitude, the hemostatic effects of BZG did not decrease significantly compared to those in the lower altitude, whereas those of ordinary gauze and QCG decreased significantly at high altitude compared to those in the lower altitude.Conclusions: Coagulation derangement after acute exposure to high altitude has negative effects on ordinary gauze and QCG but has no significant negative hemostatic effects on BZG.

Molecular Mechanisms of Mild and Severe Pneumonia: Insights from RNA Sequencing.

BACKGROUND This study aimed to uncover the molecular mechanisms underlying mild and severe pneumonia by use of mRNA sequencing (RNA-seq). MATERIAL AND METHODS RNA was extracted from the peripheral blood of patients with mild pneumonia, severe pneumonia, and healthy controls. Sequencing was performed on the HiSeq4000 platform. After filtering, clean reads were mapped to the human reference genome hg19. Differentially expressed genes (DEGs) were identified between the control group and the mild or severe group. A transcription factor-gene network was constructed for each group. Biological process (BP) terms enriched by DEGs in the network were analyzed and these genes were also mapped to the Connectivity map to search for small-molecule drugs. RESULTS A total of 199 and 560 DEGs were identified from the mild group and severe group, respectively. A transcription factor-gene network consisting of 215 nodes and another network consisting of 451 nodes were constructed in the mild group and severe group, respectively, and 54 DEGs (e.g., S100A9 and S100A12) were found to be common, with consistent differential expression changes in the 2 groups. Genes in the transcription factor-gene network for the mild group were mainly enriched in 13 BP terms, especially defense and inflammatory response (e.g., S100A8) and spermatogenesis, while the top BP terms enriched by genes in the severe group include response to oxidative stress (CCL5), wound healing, and regulation of cell differentiation (CCL5), and of the cellular protein metabolic process. CONCLUSIONS S100A9 and S100A12 may have a role in the pathogenesis of pneumonia: S100A9 and CXCL1 may contribute solely in mild pneumonia, and CCL5 and CXCL11 may contribute in severe pneumonia.

The Association between the C5263T Mutation in the Mitochondrial ND2 Gene and Coronary Heart Disease among Young Chinese Han People.

OBJECTIVE: This study aimed to investigate the genetic background of mitochondrial genes in young patients with Coronary heart disease (CHD) to provide a foundation for the early prevention of young patients with CHD. METHODS: 115 cases of young (⋜ 45 years) CHD Chinese Han patients (case group), 100 cases of older (> 45 years) Chinese Han CHD patients (experimental group) hospitalized and 100 cases of healthy people through physical examination (control group) at the General Hospital of PLA between January 2014 and December 2015 were selected. General information, clinical assessment, pedigree analysis, and mitochondrial full sequence scanning were performed. The pedigrees of one patient harbouring the C5263T mutation were recruited. Mitochondrial functional analysis including cellular reactive oxygen species (ROS) levels and mitochondrial membrane potential (MMP) were performed on pedigrees with the C5263T mutation (mutation group) and without the mutation (non-mutation group). RESULTS: The differences in biochemical tests (P > 0.05) between the case group and experimental group were not significant. The C5263T single-nucleotide mutation of the mitochondrial ND2 gene was observed in 2 young CHD patients in the case group. The premature CHD of these 2 patients followed a pattern of maternal inheritance. The mutation group (I1, II2) had higher ROS levels (4750.82 ± 1045.55 vs. 3888.58 ± 487.60, P = 0.022) and lower MMP levels (P = 0.045) than the non-mutation group (II1, III1, III2). CONCLUSION: We speculated that the mitochondrial C5263T mutation might be associated with the occurrence CHD in Chinese Han young people.

Identification of Potential Key Long Non-Coding RNAs and Target Genes Associated with Pneumonia Using Long Non-Coding RNA Sequencing (lncRNA-Seq): A Preliminary Study.

BACKGROUND This study aimed to identify the potential key long non-coding RNAs (lncRNAs) and target genes associated with pneumonia using lncRNA sequencing (lncRNA-seq). MATERIAL AND METHODS A total of 9 peripheral blood samples from patients with mild pneumonia (n=3) and severe pneumonia (n=3), as well as volunteers without pneumonia (n=3), were received for lncRNA-seq. Based on the sequencing data, differentially expressed lncRNAs (DE-lncRNAs) were identified by the limma package. After the functional enrichment analysis, target genes of DE-lncRNAs were predicted, and the regulatory network was constructed. RESULTS In total, 99 DE-lncRNAs (14 upregulated and 85 downregulated ones) were identified in the mild pneumonia group and 85 (72 upregulated and 13 downregulated ones) in the severe pneumonia group, compared with the control group. Among these DE-lncRNAs, 9 lncRNAs were upregulated in both the mild and severe pneumonia groups. A set of 868 genes were predicted to be targeted by these 9 DE-lncRNAs. In the network, RP11-248E9.5 and RP11-456D7.1 targeted the majority of genes. RP11-248E9.5 regulated several genes together with CTD-2300H10.2, such as QRFP and EPS8. Both upregulated RP11-456D7.1 and RP11-96C23.9 regulated several genes, such as PDK2. RP11-456D7.1 also positively regulated CCL21. CONCLUSIONS These novel lncRNAs and their target genes may be closely associated with the progression of pneumonia.

[Association between serum homocysteine and in-hospital death in patients with acute pulmonary embolism].

OBJECTIVE: To explore the association between serum homocysteine (Hcy) level and in-hospital death in patients with acute pulmonary embolism. METHODS: A total of 186 acute pulmonary embolism patients [ (66.8 ± 12.7) years, 89 male] hospitalized in our department between June 2008 and June 2011 were included in this prospective study. Patients were divided into high Hcy group (Hcy ≥ 15.2 µmol/L, n = 95) and low Hcy group (Hcy < 15.2 µmol/L, n = 91). Patients were followed-up for 1 year for the incidence rate of early death associated with acute pulmonary embolism. The Cox proportional hazard model was used to analyze the relationship between serum Hcy level and early death in acute pulmonary embolism patients. RESULTS: Patients were hospitalized for 1-37 days [(10 ± 6) days]. In-hospital death rate was 14.5% (27/186) and was significantly higher in high Hcy group than in low Hcy group [25.3% (24/95) vs. 3.3% (3/91) , P = 0.001]. Univariate Cox regression analysis indicated that admission heart rate, oxygen saturation, enlargement of right ventricle, Hcy ≥ 15.2 µmol/L, serum creatinine level, peak TnT level and deep venous thrombosis (P < 0.05) were independent risk factors for in-hospital death. Multivariate Cox regression analysis showed that Hcy ≥ 15.2 µmol/L (HR = 4.10, 95%CI:3.00-4.98, P = 0.017), admission heart rate (HR = 1.10, 95%CI:1.01-1.20, P = 0.031) , deep venous thrombosis (HR = 1.65, 95%CI:1.45-1.76, P = 0.034) and age (HR = 1.10, 95%CI:1.02-1.19, P = 0.010) were independent predictors of in-hospital death for acute pulmonary embolism patients. One-year follow up was finished in 142 patients (89.3%). There were 19 deaths ( 5 due to repeat pulmonary embolism, 4 due to decompensated respiratory and /or cardiac diseases, 6 due to malignant tumors, 2 due to fatal bleeding and 2 due to pneumonia) . Death rate was similar between the two groups during follow up. CONCLUSION: Higher serum homocysteine is an independent for in-hospital death for patients with acute pulmonary embolism.

[An experimental study of contrast enhanced ultrasound diagnosis of cerebral intraparenchymal hemorrhage in dogs].

OBJECTIVE: To explore the role of contrast enhanced ultrasound (CEUS) in diagnosing experimental cerebral intraparenchymal hemorrhage (IH) in dogs. METHODS: A self-control study was conducted. An IH model was reproduced by puncturing middle cerebral artery (MCA) in 12 dogs. Two-dimensional ultrasound and CEUS were conducted immediately, 30 minutes, and 1 hour after modeling, respectively, to observe the lesion echo and bleeding area. CT scans were also conducted at 1 hour after modeling, then the lesion size in CT scan was compared with that of CEUS. RESULTS: In 12 dogs IH model was reproduced successfully, and unilateral hematomas were confirmed by CT and pathological examination. Two-dimensional ultrasound of IH showed irregular high-echo area, with unclear boundary, but it was not able to show active bleeding. CEUS demonstrated active bleeding by outflow and pooling of contrast agent with obvious enhancement. CEUS of the hematoma showed perfusion deficit, with a clear boundary. The size of bleeding lesions (cm) continued to increase at 30 minutes and 1 hour after modeling (1.47±0.40, 1.76±0.45 by CEUS measuring), and demonstrated statistically significant difference comparing with the measurement of IH immediately after modeling (1.03±0.24, both P<0.01), while there was no statistically significant difference between the 30-minute and 1-hour measurements (P>0.05). Compared with the measurements between CEUS and CT at 1 hour, there was no statistically significant difference in size of bleeding lesions (1.76±0.45 vs. 1.79±0.47, P>0.05). CONCLUSION: CEUS can help determine the extent and size of IH, and the process of hematoma formation when dynamically monitoring.

Real-time ultrasound-guided external intracerebral hemorrhage drain placement.

We report a new minimally invasive technique utilizing interventional ultrasound for precise external intracerebral hemorrhage drain (EICHD) placement in pigs.

Prediction of fatal adverse prognosis in patients with fever-related diseases based on machine learning: A retrospective study.

BACKGROUND: Fever is the most common chief complaint of emergency patients. Early identification of patients at an increasing risk of death may avert adverse outcomes. The aim of this study was to establish an early prediction model of fatal adverse prognosis of fever patients by extracting key indicators using big data technology. METHODS: A retrospective study of patients' data was conducted using the Emergency Rescue Database of Chinese People's Liberation Army General Hospital. Patients were divided into the fatal adverse prognosis group and the good prognosis group. The commonly used clinical indicators were compared. Recursive feature elimination (RFE) method was used to determine the optimal number of the included variables. In the training model, logistic regression, random forest, adaboost and bagging were selected. We also collected the emergency room data from December 2018 to December 2019 with the same inclusion and exclusion criterion. The performance of the model was evaluated by accuracy, F1-score, precision, sensitivity and the areas under receiver operator characteristic curves (ROC-AUC). RESULTS: The accuracy of logistic regression, decision tree, adaboost and bagging was 0.951, 0.928, 0.924, and 0.924, F1-scores were 0.938, 0.933, 0.930, and 0.930, the precision was 0.943, 0.938, 0.937, and 0.937, ROC-AUC were 0.808, 0.738, 0.736, and 0.885, respectively. ROC-AUC of ten-fold cross-validation in logistic and bagging models were 0.80 and 0.87, respectively. The top six coefficients and odds ratio (OR) values of the variables in the Logistic regression were cardiac troponin T (CTnT) (coefficient=0.346, OR = 1.413), temperature (T) (coefficient=0.235, OR = 1.265), respiratory rate (RR) (coefficient= -0.206,OR = 0.814), serum kalium (K) (coefficient=0.137, OR = 1.146), pulse oxygen saturation (SPO2) (coefficient= -0.101, OR = 0.904), and albumin (ALB) (coefficient= -0.043, OR = 0.958). The weights of the top six variables in the bagging model were: CTnT, RR, lactate dehydrogenase, serum amylase, heartrate, and systolic blood pressure. CONCLUSIONS: The main clinical indicators of concern included CTnT, RR, SPO2, T, ALB and K. The bagging model and logistic regression model had better diagnostic performance comprehesively. Those may be conducive to the early identification of critical patients with fever by physicians.

[Vicissitude of antibiotic sensitivity of Gram negative bacilli in patients with infection in emergency departments in Beijing during 2005 to 2007].

OBJECTIVE: To investigate changes in antibiotic sensitivity of Gram negative bacilli infections among emergency patients in large hospitals in Beijing during 2005 to 2007. METHODS: Retrospective analysis of all the identified strains of Gram negative bacilli, and their sensitivity to antibiotic obtained in the emergency departments of 5 top first-class hospitals in Beijing for recent 3 years. RESULTS: Two thousand two hundred and eighty-five strains of Gram negative bacilli had taken 64.62% of all, the priority 5 of which were Pseudomonas aeruginosa (23.89%), Escherichia coli (19.91%), Acinetobacter baumanii (17.59%), Klebsiella pneumoniae (8.05%) and Staphylococcus maltophilia (3.94%). The sensitivity of commonly used antibiotics plummeted greatly in recent 3 years, especially carbapenems, cephalosporins and quinolones. The resistance of imipenem-resistant Pseudomonas aeruginosa was 17.30%, 22.53%, 31.92% respectively. The resistance of imipenem-resistant Acinetobacter baumanii was 13.87%, 12.09%, 23.56% respectively. CONCLUSION: Infectious bacteria in emergency departments in Beijing top first-class hospitals tend to show the characteristics of hospital infection. In recent years, the sensitivity of bacilli to antibiotic has dropped greatly. The situation of antibiotics resistance might be depressed.

[Protective effect of insulin-like growth factor-1 on acute lung injury induced by perfluoroisobutylene inhalation in mice].

OBJECTIVE: To observe the protective effect of insulin-like growth factor-1 (IGF-1) on acute lung injury induced by perfluoroisobutylene (PFIB) inhalation in mice. METHODS: Sixty-four male Kunming mice were randomly divided into normal control (A) group, exposed (B) group, recombinant adenoviruses 5 of IGF-1 (Ad5-IGF-1) intervention (C) group (in which Ad5-IGF-1 was injected into the trachea of the mice), blank vector control (D) group. B, C and D groups were exposed to gaseous PFIB in a flow-past whole-body exposure system. The lung index, concentration of total protein and albumin in bronchoalveolar lavage fluid (BALF), concentration of IGF-1 in serum and lung homogenate were measured. The lung pathologic changes were examined with light microscope, and ultrastructure changes in alveolar type II cells (ATII) with electron microscope. RESULTS: Compared with A group, the lung index, concentration of total protein in BALF were significantly increased in other groups, the lung index and concentration of total protein and albumin of BALF in B and D groups were prominently higher than C group (all P<0.01). The concentration of IGF-1 in serum of B and D groups was lower markedly than that of A group, and the concentration of IGF-1 in serum of C group was distinctly higher than those of A, B, D groups (all P<0.01). The concentration of IGF-1 in lung homogenate of B, C, D groups was higher than that of A group, and the concentration of IGF-1 in lung homogenate of C group was significantly higher than that of B and D groups (all P<0.01). Lung hyaline membrane formation, diffuse alveolar atelectasis, accumulation of edema fluid, red blood cell exudation, were obviously milder in C group, and changes in the ultrastructure of ATII showed a similar result. CONCLUSION: The protective effect of Ad5-IGF-1 against the toxicity of PFIB inhalation is identified. In the mice pretreated with Ad5-IGF-1 is able to significantly lower lung index, the protein concentration in BALF, and the concentration of IGF-1 in serum and lung homogenate is obviously increased. Protection of ATII may be one of the mechanisms.

[Role of contrast-enhanced ultrasound in management of splenic trauma].

OBJECTIVE: To study the role of contrast-enhanced ultrasound (CEUS) in the management of splenic trauma METHODS: Forty patients with splenic trauma underwent CEUS examination and then different management were provided according to the examination results and the general status of patients: conservative treatment (group I, n = 19); CEUS-guided injective therapy (group II, n = 6); and surgery (group III, n = 15). RESULTS: Eighteen patients were cured in group I and one patient experienced rehaemorrhagia. The haemostatic effect of CEUS-guided injective therapy was obvious in all six patients in group II. Among them, one patient experienced arteriovenous fistula, which was resolved after one week of injective therapy. Fifteen patients in Group III underwent surgery and were cured. CONCLUSION: CEUS can provide reliable information for therapy mode selection in patients with splenic trauma and can be used to guide injective therapy.

[An experimental study of a rat model with MODS as a result of trauma induced infection].

OBJECTIVE: To reproduce an animal model of multiple organ dysfunction syndrome (MODS) which was caused by two "hits" (injury and infection), to explore the potential aetiology and strategies of treatment. METHODS: The rats' extremities were crushed, resulting in severe injury with multiple closed fractures and extensive contusion of soft tissues. After 12 hours, a thirty percent total body surface area III (TBSA III) burns contaminated by Pseudomonas aeruginosa was inflicted upon the injury rats in MODS group. Rats'activities and wound appearance were observed, body weight, temperature, heart rate were recorded, and the blood lipopolysaccharide (LPS) level, the functional changes in all major organs, the occurrence rate of inflammatory reactions, the mortality of MODS and morbidity, and the pathological changes of the major organs were monitored at 24, 48, 96, 120 hours. RESULTS: To compare with pre-injury states and control group there were marked increases in alanine aminotransferase (ALT), total bilirubin (TBil), blood urea nitrogen (BUN), creatinine (Cr), aspartate aminotransferase (AST), creatine phosphokinase (CPK) contents 48 hours after the injuries (all P<0.05). When compared with the control group, there was significant difference (P<0.05). After 48 hours, there were obvious changes in pathology, the rats were in the early stage of MODS along with signs of damage to several organs. There was a typical relationship between LPS and organ functional changes. The LPS level peaked within 96 hours after the injury, the level was 8.36 folds of basic level. After 96 hours, there was correlation between LPS and the change of organ function (r=0.927 2). The incidence of MODS was 86%, and the mortality rate reached 30% in 96 hours after injury. At 120 hours after the injury, MODS was found in all the rats and the mortality rate reached 50%. CONCLUSION: This model seems to mimick the development of MODS which occurs after serious injuries followed by infection. The process of infection is coincidental with clinical picture, and LPS was released steadily with full body reaction. This animal model provides us an excellent opportunity to explore the pathogenesis and treatment of MODS after trauma.

Identification of potential key genes associated with severe pneumonia using mRNA-seq.

This study aimed to identify the potential key genes associated with severe pneumonia using mRNA-seq. Nine peripheral blood samples from patients with severe pneumonia alone (SP group, n=3) and severe pneumonia accompanied with chronic obstructive pulmonary disease (COPD; CSP group, n=3), as well as volunteers without pneumonia (control group, n=3) underwent mRNA-seq. Based on the sequencing data, differentially expressed genes (DEGs) were identified by Limma package. Following the pathway enrichment analysis of DEGs, the genes that were differentially expressed in the SP and CSP groups were selected for pathway enrichment analysis and coexpression analysis. In addition, potential genes related to pneumonia were identified based on the information in the Comparative Toxicogenomics Database. In total, 645 and 528 DEGs were identified in the SP and CSP groups, respectively, compared with the normal controls. Among these DEGs, 88 upregulated genes and 80 downregulated genes were common between the two groups. The functions of the common DEGs were similar to those of the DEGs in the SP group. In the coexpression network, the commonly downregulated genes (including ND1, ND3, ND4L, and ND6) and the commonly upregulated genes (including TSPY6P and CDY10P) exhibited a higher degree. In addition, 131 DEGs (including ND1, ND3, ND6, MIR449A and TAS2R43) were predicted to be potential pneumonia-related genes. In conclusion, the present study demonstrated that the common DEGs may be associated with the progression of severe pneumonia.

[Change in gene expression of inflammation-related genes induced in multiple organ dysfunction syndrome induced by infection of injuries in rat].

OBJECTIVE: To study the changes in inflammation-related gene expression in liver tissue during the course of multiple organ dysfunction syndrome (MODS) induced by infection of injuries and its implication. METHODS: The rats model with MODS following trauma and infection was reproduced in rat. Liver tissue was harvested. The differences of gene expressions between the simple trauma group and MODS group were detected by means of cDNA microarray. RESULTS: Comparison between the two groups, differentially expressed genes included enhanced expression of genes related both of tissue damage and repair. There was also up-regulation of expression of both inflammation-related and anti-inflammation related genes. A few genes appeared down-regulated. The differences of expression extent were significant. There were up-regulation of some genes related to apoptosis and fibrosis. CONCLUSION: Differential expressions of genes in the liver tissue include both that related to the inflammation and anti-inflammation, with down-regulation and up-regulation at the same time. There is a difference in the intensity. There is also an expression of genes related to intrinsic protection, as manifested by co-existence of systemic inflammatory response syndrome (SIRS) and compensation anti-inflammatory response syndrome (CARS) under the condition of MODS. There is an imbalance in inflammatory reaction. The simultaneous up-regulation of the tissue damage and repair related genes suggests that cellular injury is accompanied by repair in the organs during the course of MODS.

[Construction of recombinant adenovirus vector of mouse keratinocyte growth factor].

OBJECTIVE: To construct a recombinant adenovirus vector expressing keratinocyte growth factor (KGF) gene in mouse. METHODS: KGF gene amplified from mouse cDNA by polymerase chain reaction (PCR). It was reverse transcript from RNA that had been harvested from C57BL/6 mouse, and then inserted into the plasmid pShuttle-CMV to construct the shuttle plasmid pShuttle-CMV-KGF (pKGF). After linearized by restriction enzyme, the plasmid was transformed into E.Coli BJ5183 containing adenovirus backbone. The homologous recombinant pAdeasy-1-pShuttle-CMV-KGF (pAd-KGF) was identified, linearized, and then transfected into HEK293 cells using the lipofectamine TM 2000 to package the adenovirus, Adeasy-1-pShuttle-CMV-KGF (Ad-KGF), followed by further amplification, caesium chloride density gradient centrifugation purification and measurement of virus titer. Ad-GFP was used as control, and its transfection efficacy was observed. RESULTS: (1) The shuttle plasmid pKGF was proved to be successfully constructed by gene sequencing and restriction enzyme, as well as the recombinant adenovirus plasmid. (2) The cytopathic effects of HEK293 cells observed under the microscope suggested that the duplication of the virus was successful. (3) The plague titration of HEK293 cells showed virus titers were 3.0 x 10(10) pfu/ml,the concentration of which was adequate for future test in vivo or in vitro. CONCLUSION: The harvest of recombinant adenovirus vector of Ad-KGF, is the first step for the future test to investigate the effects of KGF in pulmonary diseases, and the possible gene therapy to treat pulmonary fibrosis.

Identification of miRNA biomarkers of pneumonia using RNA-sequencing and bioinformatics analysis.

Pneumonia is a lower respiratory tract infection that causes dramatic mortality worldwide. The present study aimed to investigate the pathogenesis of pneumonia and identify microRNA (miRNA) biomarkers as candidates for targeted therapy. RNA from the peripheral blood plasma of participants with pneumonia (severe, n=9; non-severe, n=9) and controls (n=9) was isolated and paired-end sequencing was performed on an Illumina HiSeq4000 system. Following the processing of raw reads, the sequences were aligned against the Genome Reference Consortium human genome assembly 38 reference genome using Bowtie2 software. Reads per kilobase of transcript per million mapped read values were obtained and the limma software package was used to identify differentially expressed miRNAs (DE-miRs). Then, DE-miR targets were predicted and subjected to enrichment analysis. In addition, a protein-protein interaction (PPI) network of the predicted targets was constructed. This analysis identified 11 key DE-miRs in pneumonia samples, including 6 upregulated miRNAs (including hsa-miR-34a and hsa-miR-455) and 5 downregulated miRNAs (including hsa-let-7f-1). All DE-miRs kept their upregulation/downregulation pattern in the control, non-severe pneumonia and severe pneumonia samples. Predicted target genes of DE-miRs in the subjects with non-severe pneumonia vs. the control and the subjects with severe pneumonia vs. the non-severe pneumonia group were markedly enriched in the adherens junction and Wnt signaling pathways. KALRN, Ras homolog family member A (RHOA), ß-catenin (CTNNB1), RNA polymerase II subunit K (POLR2K) and amyloid precursor protein (APP) were determined to encode crucial proteins in the PPI network constructed. KALRN was predicted to be a target of hsa-mir-200b, while RHOA, CTNNB1, POLR2K and APP were predicted targets of hsa-let-7f-1. The results of the present study demonstrated that hsa-let-7f-1 may serve a role in the development of cancer and the Notch signaling pathway. Conversely, hsa-miR-455 may be an inhibitor of pneumonia pathogenesis. Furthermore, hsa-miR-200b might promote pneumonia via targeting KALRN.

[Experimental study of effects of cyclooxygenase-2 inhibitor on apoptosis of hypoxic myocardial cells in vitro].

OBJECTIVE: To observe the effects of cyclooxygenase (COX)-2 inhibitor on apoptosis of hypoxic myocardial cells in vitro. METHODS: The myocardial cells were obtained from new-born Wistar rats, and were dispersed to single cell with trypsin. The cells in 4th-6th passages were randomly divided into 24 samples (every sample contained 1 x 10(5) cells): normal control group, hypoxic myocardial cells control group; hypoxic myocardial cells+NS-398 (20 micromol/L) and hypoxic myocardial cells+aspirin (100 microg/L). During culture, oxygen was replaced by N(2), and the cells were cultivated in 5% CO(2)+95% N(2) at 37 centigrade for 6 hours. Either interventional medicine or same amount of dimethyl sulphoxide was added to the cells 30 minutes before hypoxia. The expression of COX-1 and COX-2 in cultured myocardial cells in vitro was examined with Western blot. The apoptosis percentage of cells in each group was examined with flow cytometry. After centrifuging the culture medium under low temperature, 6-keto-prostaglandin F(1alpha) (6-keto-PGF(1alpha)) and thromboxane B(2) (TXB(2)) were determined with radioimmunoassay (RIA). RESULTS: There was no significant difference in the expression of COX-1 among the groups. The expression of COX-2 was higher in all acute hypoxic myocardial cells groups compared with the control group. Neither aspirin nor NS-392 inhibited the expression of COX-2. The positive percentage of apoptosis of cultured myocardial cells ranked as follow: hypoxic+NS-398 group, hypoxic +aspirin group, hypoxic control group and normal control group. The differences of apoptosis rate between hypoxic+NS-398 group and other groups were significant (all P<0.05). The levels of TXB(2) in each group of cell medium ranked in the following order: normal control group, hypoxic +NS-398 group, hypoxic+aspirin group and hypoxic control group. The difference of TXB(2) level between hypoxic+NS-398 group and hypoxic control group was significant(P<0.05). The levels of 6-keto-PGF(1alpha) in each culture medium ranked in the following order: normal control group, hypoxic+aspirin group, hypoxic+NS-398 group and hypoxic control group. The difference between hypoxic+NS-398 group and hypoxic control group was significant (P<0.05). The ratio of TXB(2)/6-keto-PGF(1alpha) showed no significant difference (P>0.05) among groups. CONCLUSION: The results suggest that acute hypoxia could directly induce cultured myocardial cells to express COX-2, but do not effect expression of COX-1. Hypoxia could elevate the level of 6-keto-PGF(1alpha) and TXB(2) in culture medium. COX-2 inhibitor (NS-398) could lessen the elevation, but it could not change the ratio between TXB(2) and 6-keto PGF(1alpha). NS-398 could increase apoptosis percentage of hypoxic myocardial cells in vitro, and the effect is independent of other inflammatory cells.

[Tumor necrosis factor-α-induced protein 8 like-2 promotes apoptosis of CD4+ T lymphocytes in mice with severe burn injury].

OBJECTIVE: To investigate the effect of tumor necrosis factor-α-induced protein 8 like-2 (TIPE2) on apoptosis of CD4+ T lymphocytes in a murine model of severe burn injury. METHODS: A total of 140 male mice were randomly allocated into 6 groups. Small RNA interference technique was used to construct a siTIPE2-overexpressing lentivirus, and severe burn injury models were established in the mice. CD4+ T cells were purified from spleen of the mice, and the expressions of TIPE2, Smad2/Smad3, P-Smad2/P-Smad3 and Bcl-2/Bimprotein in CD4+ Tregs were detected. The changes in mitochondrial membrane potential and cytochrome C in CD4+ T cells were detected, and the activities of caspase-3, caspase-8, and caspase-9 were analyzed. RESULTS: Down-regulation of TIPE2 promoted the apoptosis of CD4+ T lymphocytes in siTIPE2-burn group, in which the protein expressions of P-smad2/P-Smad3 decreased, Bcl-2 expression increased and Bim expression decreased significantly as compared with the other groups (P<0.01 or 0.05). The mitochondrial membrane potential and cytochrome C expression in CD4+ T cells were down-regulated in siTIPE2-burn group (P<0.05) with a lowered caspase-3 activity compared with TIPE2-burn group (P<0.01) and decreased caspase-8 and caspase-9 compared with the other groups (P<0.05). The apoptosis rate was the highest in TIPE2-burn group, whose Smad2/Smad3 was higher than that in the sham group (P<0.05) and the expression of P-smad2/P-Smad3 significantly increased compared with the other groups (P<0.05). In TIPE2-burn group, the mitochondrial membrane potential in CD4+ T cells was decreased (P<0.01), the expression of cytochrome C increased markedly (P<0.01), and the activities of caspase-3, caspase-8, and caspase-9 were all obviously higher than those in the other groups (P<0.05). CONCLUSION: As an important immunoregulatory molecule, TIPE2 can promote the apoptosis of CD4+T lymphocyte in mice with sever burn injury.