Design, modeling, and testing of a one degree of freedom manipulator with three-stage amplification mechanism.

The one degree of freedom (1-DOF) manipulator with nano-resolution is a significant component in the micro-/nano-manipulation. In order to simultaneously achieve a large stroke and high precision, a piezo-driven 1-DOF flexure-based manipulator consisting of an enhanced double Scott-Russell mechanism (EDSRM), a lever type mechanism, and a Z-shaped mechanism is proposed in this paper. Analytical models are developed to examine the kinetostatic and dynamic properties of the manipulator. A finite element analysis is further performed to evaluate the characteristics of the EDSRM and the complete manipulator. The prototype is fabricated on monolithic AL7075, and various experimental tests have been carried out to investigate the correctness of the modeling. The experimental results show that the proposed manipulator has a satisfactory amplification ratio, static stability, and dynamic performance.

Elevated circulating miR-494 in plasma of children with enterovirus 71 induced hand, foot, and mouth disease and its potential diagnostic value.

Hand, foot, and mouth disease (HFMD) is a common pediatric disease, whose outcome depends of the enterovirus genotype infecting the patient. The present study is focused on the potential diagnostic value and the role of circulating microRNA-494 (miR-494) in enterovirus 71-induced more severe form of HFMD. We included 102 children with enterovirus 71 (EV71)-induced HFMD, 42 coxsackievirus A16 (CA16)-induced HFMD and 102 healthy controls. The plasma and serum samples were collected. The expression level of circulating miR-494 was determined by RT-PCR method. Moreover, ROC curve has been drawn to evaluate the sensitivity and specificity of circulating miR-494 for the diagnosis of EV71-induced HFMD. Furthermore, the correlation between the circulating miR-494 and the levels of interleukin 6 (IL-6), interleukin 4 (IL-4) and interferon γ (IFN-γ) in the serum of patients were analyzed. Circulating miR-494 was significantly increased in plasma of children with EV71-induced HFMD compared with the healthy children or CA16-induced HFMD, and level of miR-494 in the EV71 severe group was significantly higher than the EV71 mild group. Moreover, results of ROC analysis suggested that miR-494 is a sensitive biomarker to distinguish EV71 patients from healthy controls and CA16 patients. Furthermore, IL-6 and IFN-γ were elevated in serum of patients with EV71-induced HFMD and the level of circulating miR-494 in patients with EV71-induced HFMD was positively correlated with the serum levels of both IL-6 and IFN-γ, respectively. Circulating miR-494 was abnormally up-regulated in plasma of the children with EV71-induced HFMD, and miR-494 may serve as potential biomarker for the diagnosis and treatment of the disease. Keywords: miR-494; HFMD; biomarker; enterovirus 71; inflammation.

LncRNA LINP1 promotes proliferation and inhibits apoptosis of gastric cancer cells by repressing RBM5.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "LncRNA LINP1 promotes proliferation and inhibits apoptosis of gastric cancer cells by repressing RBM5, by X.-C. Lu, H.-Y. Zhou, J. Wu, Y. Jin, X.-M. Yao, X.-Y. Wu, published in Eur Rev Med Pharmacol Sci 2020; 24 (1): 137-144-DOI: 10.26355/eurrev_202001_19904-PMID: 31957826" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/19904.

Long non-coding RNA AB073614 promotes metastasis of gastric cancer cells by upregulating IGF-2.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long non-coding RNA AB073614 promotes metastasis of gastric cancer cells by upregulating IGF-2, by X.-Y. Wu, H.-Y. Zhou, X.-M. Yao, X.-D. Chen, J. Wu, X.-C. Lu, published in Eur Rev Med Pharmacol Sci 2020; 24 (1): 145-150-DOI: 10.26355/eurrev_202001_19905-PMID: 31957827" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/19905.

[Exploration of omics mechanism and drug prediction of coronavirus-induced heart failure based on clinical bioinformatics].

Objective: Present study investigated the mechanism of heart failure associated with coronavirus infection and predicted potential effective therapeutic drugs against heart failure associated with coronavirus infection. Methods: Coronavirus and heart failure were searched in the Gene Expression Omnibus (GEO) and omics data were selected to meet experimental requirements. Differentially expressed genes were analyzed using the Limma package in R language to screen for differentially expressed genes. The two sets of differential genes were introduced into the R language cluster Profiler package for gene ontology (GO) and Kyoto gene and genome encyclopedia (KEGG) pathway enrichment analysis. Two sets of intersections were taken. A protein interaction network was constructed for all differentially expressed genes using STRING database and core genes were screened. Finally, the apparently accurate treatment prediction platform (EpiMed) independently developed by the team was used to predict the therapeutic drug. Results: The GSE59185 coronavirus data set was searched and screened in the GEO database, and divided into wt group, ΔE group, Δ3 group, Δ5 group according to different subtypes, and compared with control group. After the difference analysis, 191 up-regulated genes and 18 down-regulated genes were defined. The GEO126062 heart failure data set was retrieved and screened from the GEO database. A total of 495 differentially expressed genes were screened, of which 165 were up-regulated and 330 were down-regulated. Correlation analysis of differentially expressed genes between coronavirus and heart failure was performed. After cross processing, there were 20 GO entries, which were mainly enriched in virus response, virus defense response, type Ⅰ interferon response, γ interferon regulation, innate immune response regulation, negative regulation of virus life cycle, replication regulation of viral genome, etc. There were 5 KEGG pathways, mainly interacting with tumor necrosis factor (TNF) signaling pathway, interleukin (IL)-17 signaling pathway, cytokine and receptor interaction, Toll-like receptor signaling pathway, human giant cells viral infection related. All differentially expressed genes were introduced into the STRING online analysis website for protein interaction network analysis, and core genes such as signal transducer and activator of transcription 3, IL-10, IL17, TNF, interferon regulatory factor 9, 2'-5'-oligoadenylate synthetase 1, mitogen-activated protein kinase 3, radical s-adenosyl methionine domain containing 2, c-x-c motif chemokine ligand 10, caspase 3 and other genes were screened. The drugs predicted by EpiMed's apparent precision treatment prediction platform for disease-drug association analysis were mainly TNF-α inhibitors, resveratrol, ritonavir, paeony, retinoic acid, forsythia, and houttuynia cordata. Conclusions: The abnormal activation of multiple inflammatory pathways may be the cause of heart failure in patients after coronavirus infection. Resveratrol, ritonavir, retinoic acid, amaranth, forsythia, houttuynia may have therapeutic effects. Future basic and clinical research is warranted to validate present results and hypothesis.

LncRNA LINP1 promotes proliferation and inhibits apoptosis of gastric cancer cells by repressing RBM5.

OBJECTIVE: Recent studies have revealed that long non-coding RNAs (lncRNAs) play a crucial role in tumor progression. Gastric cancer (GC) is one of the common types of malignancies worldwide. This study aimed to identify the exact function of lncRNA LINP1 in the progression of GC. PATIENTS AND METHODS: LINP1 expression in paired cancer tissues and adjacent normal tissues of GC patients was detected by Real-Time quantitative Polymerase Chain Reaction (RT-qPCR). The effect of LINP1 silence on proliferation and apoptosis of GC cells was detected. Meanwhile, the underlying mechanism of LINP1 function was explored by RT-qPCR and Western blot assay. Furthermore, tumor formation assay was performed to examine the ability of LINP1 in tumor formation in vivo. RESULTS: LINP1 expression was remarkably up-regulated in GC tissues compared with adjacent normal tissues. The growth ability of GC cells was significantly inhibited after silencing of LINP1 in vitro. Besides, the apoptosis of GC cells was markedly induced after silencing of LINP1. The silence of LINP1 significantly up-regulated the expression of RBM5 in GC cells. Meanwhile, RBM5 expression in GC tissues was remarkably lower than that of the adjacent normal tissues. Furthermore, tumor formation assay showed that knockdown of LINP1 markedly inhibited tumor formation in vivo. CONCLUSIONS: These results suggested that LINP1 could down-regulate RBM5. Meanwhile, LINP1 remarkably promoted growth ability and suppressed apoptosis of GC in vitro and in vivo. Our findings might provide a novel regulator and therapeutic strategy for GC patients.

Long non-coding RNA AB073614 promotes metastasis of gastric cancer cells by upregulating IGF-2.

OBJECTIVE: Recently, long non-coding RNAs (lncRNAs) have been widely studied for their vital roles in human diseases. In this study, we investigated the effect of lncRNA AB073614 on the metastasis of gastric cancer (GC), and explored the possible underlying mechanism. PATIENTS AND METHODS: AB073614 expression in GC tissue samples was detected by Real-time quantitative polymerase chain reaction (RT-qPCR). The roles of AB073614 in GC metastasis were identified through wound healing assay and transwell assay, respectively. Moreover, RT-qPCR and Western blot assay were used to explore the potential mechanism. RESULTS: AB073614 expression level in GC samples was significantly higher than that of adjacent ones. Besides, the migration and invasion of GC cells were obviously repressed after AB073614 was knocked down. After AB073614 was knocked down in vitro, the mRNA and protein expressions of insulin-like growth factor 2 (IGF-2) was remarkably down-regulated. Furthermore, a negative correlation was found between the expression level of IGF-2 and AB073614 in GC tissues. CONCLUSIONS: AB073614 could promote GC cell migration and invasion via up-regulating IGF-2. Our findings might provide a potential therapeutic target for GC patients.

[Effect of comprehensive AIDS intervention among men aged 50 or over who had non-marital sexual behavior].

Objective: To evaluate the effectiveness of AIDS intervention programs on men aged 50 or over and having had non-marital sexual behavior. Methods: A community-based intervention/experimental and based on individual level study was adopted. Stratified sampling method was used. 12 townships/streets in Fuyang district of Hangzhou were identified as intervention or control group (six research sites each). All of the subjects in the township (street) were included. The inclusion criteria of study objects would include men aged 50 or older who reported having unmarried sex in the last year. Estimated sample size was 290, with each 145 in the intervention group and the control group. All the intervention group participants were provided with a total of 4 intervention-related items (knowledge and education on AIDS prevention, information radiation and behavioral change, broadcast expert lectures), every 3 months, for 12 month, the main evaluation indicators would include: incidence of non-marital sex and commercial sex in the last year, condom use when having non-marital sex in the last episode. Results: A total of 312 subjects were recruited. 300 of them completed the baseline study while 284 of them completed the follow-up survey. Among the subjects who had undergone the baseline study, the average age was (65.58±7.89), 71.33% were married or cohabiting with someone, 52.00% having had primary school education. After the implementation of intervention programs, the incidence of non-marital sex dropped to 59.42% (82/138) and the incidence of commercial sex dropped from 79.73% (118/148) to 55.07% (76/138). Condom use rate in the last non-marital sexual contact increased from 19.59% (29/148) to 51.22% (42/82). In the control group, the incidence of non-marital sex in the year before dropped to 74.66% (109/146) and the incidence of commercial sex dropped from 91.45% (139/152) to 72.60% (106/146). Rates of condom use during the last non-marital sexual contact dropped from 32.89% (50/152) to 31.19% (34/109). Statistically, there were significant differences appeared between the two groups on the incidence of non-marital sex in the past year (χ(2)=7.48, P=0.008), the incidence of commercial sex in the last year (χ(2)=9.47, P=0.003) and the rate of condom use in the last sex experience (χ(2)=7.83, P=0.007). Conclusions: Results from this intervention study showed that: in the intervention group, both the incidence rates of non-marital or commercial sex had reduced, together with the increase of condom use in non-marital sex in the last sexual experience. Intervention strategies that involving knowledge and education on AIDS prevention, information radiation and behavioral change, broadcasting lectures by experts etc. were all proved effective.

Long noncoding RNA LINC00265 predicts the prognosis of acute myeloid leukemia patients and functions as a promoter by activating PI3K-AKT pathway.

OBJECTIVE: Mounting evidence suggests that long noncoding RNAs (lncRNAs) function in multiple cancers. This study aimed to determine the expression, clinical significance, and possible biological function of a novel lncRNA LINC00265 in acute myeloid leukemia (AML). PATIENTS AND METHODS: The expression levels of LINC00265 were systematically evaluated in TCGA datasets. RT-PCR was performed to examine the expression level of LINC00265 in bone marrow and serum obtained from AML patients and healthy controls. The clinical data were interpreted by x2 test, Kaplan-Meier analyses, univariate analysis, and multivariate analysis. The functional role of LINC00265 was verified using cell experiments. Western blotting was used to examine the modulatory effect of LINC00265 on AKT/PI3K pathway in AML. RESULTS: LINC00265 was significantly highly expressed in the bone marrow and serum of AML patients. High serum LINC00265 was significantly associated with FAB classification and cytogenetics. ROC analyses showed that serum LINC00265 levels were reliable in distinguishing patients with AML from normal controls. Clinical assay indicated that AML patients with higher serum LINC00265 expression suffered poorer overall survival. Functionally, overexpression of LINC00265 suppressed the capability of proliferation, migration and invasion in AML cell lines. By using Western blot, we further illustrated that LINC00265 activated PI3K/AKT signaling in AML cell lines. CONCLUSIONS: Our findings not only demonstrated that LINC00265 contributes to AML proliferation, migration and invasion via modulation of PI3K/AKT signaling, but also suggested the potential value of LINC00265 as a clinical prognostic and a diagnostic marker for AML.

[Gene expression profile analysis involved in pathogenesis of allergic rhinitis by using bioinformatics method as a novel way of drug screening].

Objective: To analyze the dynamic changes of the clinical features of chronic rhinosinusitis in recent 10 years, so as to deeply understand the characteristics of chronic rhinosinusitis, and to provide new ideas for treatment of chronic rhinosinusitis. Method: This retrospective study was performed in patients who were diagnosed as chronic rhinosinusitis and enrolled. General information, clinical examination and pathological results were all collected, then patients' age, gender, the incidence of asthma and allergic rhinitis, peripheral eosinophil percentage, olfactory dysfunction and pathological results were statistically analyzed. Result: 1 955 patients who were diagnosed as chronic rhinosinusitis(CRS) were enrolled in this study, including 570 patients in 2006, 583 patients in 2010, and 802 patients in 2015. There were no obvious changes of age structure in these patients in three years. And there was no significant change in sex ratio as well. The proportions of patients with CRS concomitant with asthma were obviously increased in 10 years, which was 3.51% in 2006, 7.55% in 2010, and 17.58% in 2015. The proportions of patients with allergic rhinitis were also increased, which was 10.35% in 2006, 8.75% in 2010, and 14.09% in 2015. Peripheral eosinophil ratio was increased significantly in these patients after 2010. The proportions of ECRS in CRS were elevated in 2015 and almost doubled compared to 2006. Olfactory dysfunction increased significantly in 2015. Conclusion: In recent 10 years, there were obvious changes of clinical features of CRS. The proportion of patients with CRS concomitant with asthma showed a gradual increasing trend. ECRS significantly increased than it was 10 years ago. Olfactory dysfunction also increased significantly. In order to improve the therapeutic effect of CRS, it is necessary to strengthen the treatment of upper and lower airway inflammation related with eosinophil.

MiR-222 promotes proliferation, migration and invasion of lung adenocarcinoma cells by targeting ETS1.

OBJECTIVE: The lung adenocarcinoma is a type of lung cancer. This research is to investigate the effects of miR-222 on the proliferation, migration and invasion of the lung adenocarcinoma cells. MATERIALS AND METHODS: At the beginning, MiR-222 and the controls were transfected to the lung adenocarcinoma cell line A549 for CCK-8 proliferation, transwell migration and Matrigel invasion, and then observed the effect of miR-222 on the proliferation, migration and invasion of lung adenocarcinoma cells. The miR-222 target was regulated by ETS1 downwards to participate in the regulation of the process by using the luciferase reporter assay, the Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and the Western blotting. RESULTS: According to CCK-8 proliferation assay, the Transwell migration and the Matrigel invasion assay, it discovered that MiR-222 can promote the proliferation, migration and invasion of the lung adenocarcinoma cells. Luciferase reporter assay, RT-qPCR and Western blot assay showed that miR-222 could regulate the expression of ETS1 downwards and ETS1 participated in the regulation of the process CONCLUSIONS: ETS1 promotes proliferation, migration and invasion of lung adenocarcinoma cells by targeting the regulated miR-222 downwards.

Statin-treated patients with aneurysmal subarachnoid haemorrhage: a meta-analysis.

OBJECTIVE: The cerebral vasospasm, delayed ischemic neurological deficit (DIND), mortality and poor neurological outcome induced by aneurysmal subarachnoid haemorrhage (SAH) remain the major causes of morbidity and mortality in aneurysmal SAH patients. The effects of statin-treated for aneurysmal SAH patients were not comprehensively assessed. PATIENTS AND METHODS: A systematically literature search was conducted in PubMed, EMBASE, ScienceDirect and Web of Science to identify relevant studies update to March 2015. Data were extracted and appraised independently by two authors. Moreover, fixed or random effects models were applied to calculate pooled results based on the degree of heterogeneity. RESULT: Nine RCTs and three observational studies with a total of 1957 patients met the inclusion criteria. The results showed that statin treatment was not associated with a decrease in the occurrence of DIND (RR: 0.81, 95% CI: 0.66-1.00, p = 0.05), mortality (RR: 0.90, 95% CI: 0.69-1.18, p = 0.46) and poor neurological outcome (RR: 1.02, 95% CI: 0.86-1.20, p = 0.84), nonetheless, had a potential effect on reducing the incidence of vasospasm (RR: 0.77, 95% CI: 0.66-0.89, p = 0.0006). CONCLUSIONS: This meta-analysis indicated that the use of statins decreases the occurrence of cerebral vasospasm, whereas did not support a beneficial effect of statins on the occurrence of DIND, death or poor neurological outcomes in patients with aneurysmal SAH.

[Observation of the clinical efficacy of tigecycline for treatment of ventilator-associated pneumonia in critically ill elderly patients].

OBJECTIVE: To evaluate the efficacy of tigecycline for treatment of ventilator-associated pneumonia in critically ill elderly patients. METHODS: Data of critically ill elderly patients with ventilator-associated pneumonia treated with tigecycline in the intensive care unit was collected from June 2011 to March 2014 in this retrospective study, to evaluated the clinical efficacy of tigecycline. RESULTS: A total of 79 patients (83.5% male) were included, the mean age was 84 years old (rang, 65 years to 100 years old). Acinetobacter baumannii (39.1%), Pseudomonas aeruginosa (35.0%) and Klebsiella pneumonia (23.8%) were the most common pathogens.All patients were treated with tigecycline, 54.4% combined with other antimicrobial agents, 35.4% treated with double dose of tigecycline, and the mean course of antibiotic treatment was 9 days (range, 2 days to 22 days). After treatment, clinical success were recorded in 44 patients (55.7%), clinical failure were recorded in 29 patients, clinical uncertainty were recorded in 6 patients.28 days after treatment, patients' overall mortality was 39.0%.The clinical success rates were associated with acute physiology and chronic health evaluation (APACHE) Ⅱ score less than 15 (the clinical success rates were 72.2% and 41.9% in patients with APACHE Ⅱ score<15 and APACHE Ⅱ score≥15, respectively; P=0.007); treated with double dose of tigecycline (71.4% vs 47.1%, P=0.037) or combination regimens were also had significant difference (67.4% vs 41.7%, P=0.022). CONCLUSIONS: Treatment of tigecycline combined with other antimicrobial agents and double dose of tigecycline may both can improve clinical efficacy in critically ill elderly patients with ventilator-associated pneumonia.

Association between HLA rs3129882 polymorphism and Parkinson's disease: a meta-analysis.

OBJECTIVE: As one of potential candidate genes for the risk of Parkinson's disease (PD), the HLA-DRA/PARK18 (rs3129882, A > G) gene has been studied extensively. However, direct evidence for the genetic association studies between PD and rs3129882 remains inconclusive. The aim of our meta-analysis was to determine a more reliable association between the rs3129882 and PD. MATERIALS AND METHODS: Comprehensive search strategy was used for electronic searches through PubMed, Elsevier, Springer Link, CNKI (Chinese National Knowledge Infrastructure) and WanFang (Chinese) databases to evaluate the association between rs3129882 and PD risk. Data were extracted and the odd ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. Finally, we performed a meta-analysis of 13 appropriate papers by using a total of 11951 patients and 11902 controls. RESULTS: The meta-analysis showed no significant association between rs3129882 and PD risk in all four models (the allele model, dominant model, homozygote model and the recessive model). In allele model, the result was OR = 1.043 (95% CI = 0.978, 1.113). Moreover, this association remained no significant in the subgroup analysis stratified by ethnicity. CONCLUSIONS: In current meta-analysis, no significant association was found for rs3129882 and PD risk. And more well-designed primary researches will be needed to further evaluate the interaction of rs3129882 polymorphism and the susceptibility of PD.

Selective brain cooling in rats ameliorates intracerebral hemorrhage and edema caused by penetrating brain injury: possible involvement of heme oxygenase-1 expression.

Brain edema formation associated with trauma-induced intracerebral hemorrhage (ICH) is a clinical complication with high mortality. Studies have shown that heme oxygenase-1 (HO-1) plays an important role in ICH-induced brain edema. In order to understand the role of HO-1 in the protective effect of selective brain cooling (SBC), we investigated the time course of HO-1 changes following penetrating ballistic-like brain injury (PBBI) in rats. Samples were collected from injured and control animals at 6, 24, 48, and 72 h, and 7 days post-injury to evaluate HO-1 expression, heme concentration, brain water content, and immunohistochemistry (IHC). Following a 10% frontal PBBI, HO-1 mRNA and protein was increased at all time points studied, reaching maximum expression levels at 24-48 h post-injury. An increase in the heme concentration and the development of brain edema coincided with the upregulation of HO-1 mRNA and protein during the 7-day post-injury period. SBC significantly decreased PBBI-induced heme concentration, attenuated HO-1 upregulation, and concomitantly reduced brain water content. These results suggest that the neuroprotective effects of SBC may be partially mediated by reducing the heme accumulation, which reduced injury-mediated upregulation of HO-1, and in turn ameliorated edema formation. Collectively, these results suggest a potential value of HO-1 as a diagnostic and/or therapeutic biomarker in hemorrhagic brain injury.

Synergism of human amnion-derived multipotent progenitor (AMP) cells and a collagen scaffold in promoting brain wound recovery: pre-clinical studies in an experimental model of penetrating ballistic-like brain injury.

One of the histopathological consequences of a penetrating ballistic brain injury is the formation of a permanent cavity. In a previous study using the penetrating ballistic-like brain injury (PBBI) model, engrafted human amnion-derived multipotent progenitor (AMP) cells failed to survive when injected directly in the injury tract, suggesting that the cell survival requires a supportive matrix. In this study, we seated AMP cells in a collagen-based scaffold, injected into the injury core, and investigated cell survival and neuroprotection following PBBI. AMP cells suspended in AMP cell conditioned medium (ACCS) or in a liquefied collagen matrix were injected immediately after a PBBI along the penetrating injury tract. Injured control rats received only liquefied collagen matrix. All animals were allowed to survive two weeks. Consistent with our previous results, AMP cells suspended in ACCS failed to survive; likewise, no collagen was identified at the injury site when injected alone. In contrast, both AMP cells and the collagen were preserved in the injury cavity when injected together. In addition, AMP cells/collagen treatment preserved some apparent brain tissue in the injury cavity, and there was measurable infiltration of endogenous neural progenitor cells and astrocytes into the preserved brain tissue. AMP cells were also found to have migrated into the subventricular zone and the corpus callosum. Moreover, the AMP cell/collagen treatment significantly attenuated the PBBI-induced axonal degeneration in the corpus callosum and ipsilateral thalamus and improved motor impairment on rotarod performance. Overall, collagen-based scaffold provided a supportive matrix for AMP cell survival, migration, and neuroprotection.

Human amnion-derived multipotent progenitor cell treatment alleviates traumatic brain injury-induced axonal degeneration.

To identify a viable cell source with potential neuroprotective effects, we studied amnion-derived multipotent progenitor (AMP) cells in a rat model of penetrating ballistic-like brain injury (PBBI). AMP cells were labeled with fluorescent dye PKH26 and injected in rats immediately following right hemispheric PBBI or sham PBBI surgery by ipsilateral i.c.v. administration. At 2 weeks post-injury, severe necrosis developed along the PBBI tract and axonal degeneration was prominent along the corpus callosum (cc) and in the ipsilateral thalamus. Injected AMP cells first entered the subventricular zone (SVZ) in both sham and PBBI rats. Further AMP cell migration along the cc only occurred in PBBI animals. No significant difference in injury volume was observed across all treatment groups. In contrast, treatment with AMP cells significantly attenuated axonal degeneration in both the thalamus and the cc. Interestingly, PKH26-labeled AMP cells were detected only in the SVZ and the cc (in parallel with the axonal degeneration), but not in the thalamus. None of the labeled AMP cells appeared to express neural differentiation, as evidenced by the lack of double labeling with nestin, S-100, GFAP, and MAP-2 immunostaining. In conclusion, AMP cell migration was specifically induced by PBBI and requires SVZ homing, yet the neuroprotective effect of intracerebral ventrical treatment using AMP cells was not limited to the area where the cells were present. This suggests that the attenuation of the secondary brain injury following PBBI was likely to be mediated by mechanisms other than cell replacement, possibly through delivery or sustained secretion of neurotrophic factors.

P43/pro-EMAPII: a potential biomarker for discriminating traumatic versus ischemic brain injury.

To gain additional insights into the pathogenic cellular and molecular mechanisms underlying different types of brain injury (e.g., trauma versus ischemia), recently attention has focused on the discovery and study of protein biomarkers. In previous studies, using a high-throughput immunoblotting (HTPI) technique, we reported changes in 29 out of 998 proteins following acute injuries to the rat brain (penetrating traumatic versus focal ischemic). Importantly, we discovered that one protein, endothelial monocyte-activating polypeptide II precursor (p43/pro-EMAPII), was differentially expressed between these two types of brain injury. Among other functions, p43/pro-EMAPII is a known pro-inflammatory cytokine involved in the progression of apoptotic cell death. Our current objective was to verify the changes in p43/pro-EMAPII expression, and to evaluate the potentially important implications that the differential regulation of this protein has on injury development. At multiple time points following either a penetrating ballistic-like brain injury (PBBI), or a transient middle cerebral artery occlusion (MCAo) brain injury, tissue samples (6-72 h), CSF samples (24 h), and blood samples (24 h) were collected from rats for analysis. Changes in protein expression were assessed by Western blot analysis and immunohistochemistry. Our results indicated that p43/pro-EMAPII was significantly increased in brain tissues, CSF, and plasma following PBBI, but decreased after MCAo injury compared to their respective sham control samples. This differential expression of p43/pro-EMAPII may be a useful injury-specific biomarker associated with the underlying pathologies of traumatic versus ischemic brain injury, and provide valuable information for directing injury-specific therapeutics.

Modeling cerebral ischemia in neuroproteomics.

Protein changes induced by traumatic or ischemic brain injury can serve as diagnostic markers as well as therapeutic targets for neuroprotection. The focus of this chapter is to provide a representative overview of preclinical brain injury and proteomics analysis protocols for evaluation and discovery of novel biomarkers. Detailed surgical procedures have been provided for inducing MCAo and implantation of chronic indwelling cannulas for drug delivery. Sample collection and tissue processing techniques for collection of blood, CSF, and brain are also described including standard biochemical methodology for the proteomic analysis of these tissues.The dynamics of proteomic analysis is a multistep process comprising sample preparation, separation, quantification, and identification of proteins. Our approach is to separate proteins first by two-dimensional gel electrophoresis according to charge and molecular mass. Proteins are then fragmented and analyzed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Identification of proteins can be achieved by comparing the mass-to-charge data to protein sequences in respective databases.

Detection of protein biomarkers using high-throughput immunoblotting following focal ischemic or penetrating ballistic-like brain injuries in rats.

PRIMARY OBJECTIVE: Recent efforts have been aimed at developing a panel of protein biomarkers for the diagnosis/prognosis of the neurological damage associated with acute brain injury. METHODS AND PROCEDURES: This study utilized high-throughput immunoblotting (HTPI) technology to compare changes between two animal models of acute brain injury: penetrating ballistic-like brain injury (PBBI) which mimics the injury created by a gunshot wound and transient middle cerebral artery occlusion (MCAo) which is a model of stroke. Brain and blood were collected at 24-hours post-injury. MAIN OUTCOMES AND RESULTS: This study identified the changes in 18 proteins following PBBI and 17 proteins following MCAo out of a total of 998 screened proteins. Distinct differences were observed between the two models: five proteins were up- or down-regulated in both models, 23 proteins changed in only one model and one protein was differentially expressed. Western blots were used to verify HTPI results for selected proteins with measurable changes observed in both blood and brain for the proteins STAT3, Tau, PKA RII beta, 14-3-3 epsilon and p43/EMAPII. CONCLUSIONS: These results suggest distinct post-injury protein profiles between brain injury types (traumatic vs. ischemic) that will facilitate strategies aimed at the differential diagnosis and prognosis of acute brain injury.