Transplantation of human placental chorionic plate-derived mesenchymal stem cells for repair of neurological damage in neonatal hypoxic-ischemic encephalopathy

Neonatal hypoxic-ischemic encephalopathy is often associated with permanent cerebral palsy,neurosensory impairments,and cognitive deficits,and there is no effective treatment for complications related to hypoxic-ischemic encephalopathy.The therapeutic potential of human placental chorionic plate-derived mesenchymal stem cells for various diseases has been explored.However,the potential use of human placental chorionic plate-derived mesenchymal stem cells for the treatment of neonatal hypoxic-ischemic encephalopathy has not yet been investigated.In this study,we injected human placental chorionic plate-derived mesenchymal stem cells into the lateral ventricle of a neonatal hypoxic-ischemic encephalopathy rat model and observed significant improvements in both cognitive and motor function.Protein chip analysis showed that interleukin-3 expression was significantly elevated in neonatal hypoxic-ischemic encephalopathy model rats.Following transplantation of human placental chorionic plate-derived mesenchymal stem cells,interleukin-3 expression was downregulated.To further investigate the role of interleukin-3 in neonatal hypoxic-ischemic encephalopathy,we established an in vitro SH-SY5Y cell model of hypoxic-ischemic injury through oxygen-glucose deprivation and silenced interleukin-3 expression using small interfering RNA.We found that the activity and proliferation of SH-SY5Y cells subjected to oxygen-glucose deprivation were further suppressed by interleukin-3 knockdown.Furthermore,interleukin-3 knockout exacerbated neuronal damage and cognitive and motor function impairment in rat models of hypoxic-ischemic encephalopathy.The findings suggest that transplantation of hpcMSCs ameliorated behavioral impairments in a rat model of hypoxic-ischemic encephalopathy,and this effect was mediated by interleukin-3-dependent neurological function.

Differential expression of plasma cytokines in sepsis patients and their clinical implications

BACKGROUND Sepsis,which is characterized by acute systemic inflammation and is associated with high rates of morbidity and mortality,presents a significant challenge in health care.Some scholars have found that the sequential organ failure assessment(SOFA)and quick SOFA scores are not ideal for predicting severe sepsis and mortality.Microbial culture takes a long time(2-3 d)and provides no information for early diagnosis and treatment.Therefore,new diagnostic methods for sepsis need to be explored.AIM To assess cytokine levels in the plasma of sepsis patients and identify potential biomarkers for diagnosing sepsis.METHODS Ten sepsis patients admitted to the emergency department within 24 h of onset were enrolled as the observation group,whereas ten noninfected patients served as the control group.Of the 10 noninfected patients,9 hypertension combined with cerebral infarction,1 patients with vertiginous syndrome.Plasma Cytokines were measured using the Bio-Plex Pro^(TM)Human Chemokine Panel 40-plex.Differentially expressed cytokines in plasma of sepsis and nonsepsis patients were analyzed using Gene Ontology(GO)functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses.RESULTS Interleukin(IL)-16,granulocyte-macrophage granulocyte-macrophage colony-stimulating factor(GM-CSF),CX3CL1,CXCL9,CXCL16,CCL25,and CCL23 plasma levels were significantly increased in sepsis patients.GO analysis revealed that these cytokines were mainly associated with cellular structures such as intermediates,nuclear plaques,adhesion plaques,lateral plasma membranes,and cell matrix junctions.These genes were involved in various molecular functions,such as cytokine activity,receptor ligand activity,and signal receptor activator activity,contributing to various biological functions,such as leukocyte chemotaxis,migration,and chemotaxis.KEGG analysis indicated involvement in cytokine cytokine receptor interactions,chemokine signaling pathways,virus–protein interactions with cytokines and cytokine receptors,and the tumor necrosis factor signaling pathway.CONCLUSION Elevated serum levels of IL-16,GM-CSF,CX3CL1,CXCL9,CXCL16,CCL25,and CCL23 in sepsis patients suggest their potential as diagnostic biomarkers for sepsis.

Study on the Simultaneously Quantitative Detection for β-Lactoglobulin and Lactoferrin of Cow Milk by Using Protein Chip Technique

Objective To research a protein chip method which can simultaneously quantitative detect β-Lactoglobulin （β-L） and Lactoferrin （Lf） at one time. Methods Protein chip printer was used to print both anti-β-L antibodies and anti-Lf antibodies on each block of protein chip. And then an improved sandwich detection method was applied while the other two detecting antibodies for the two antigens were added in the block after they were mixed. The detection conditions of the quantitative detection for simultaneous measurement of β-L and Lf with protein chip were optimized and evaluated. Based on these detected conditions, two standard curves of the two proteins were simultaneously established on one protein chip. Finally, the new detection method was evaluated by using the analysis of precision and accuracy. Results By comparison experiment, mouse monoclonal antibodies of the two antigens were chosen as the printing probe. The concentrations of β-L and Lf probes were 0.5 mg/mL and 0.5 mg/mL, respectively, while the titers of detection antibodies both of β-L and Lf were 1：2,000. Intra- and inter-assay variability was between 4.88% and 38.33% for all tests. The regression coefficients of protein chip comparing with ELISA for β-L and Lf were better than 0.734, and both of the two regression coefficients were statistically significant （r = 0.734, t = 2.644, P = 0.038; and r = 0.774, t = 2.998, P = 0.024）. Conclusion A protein chip method of simultaneously quantitative detection for β-L and Lf has been established and this method is worthy in further application.

Weak cation exchange 2 protein chip for detecting differentially expressed brainstem proteins in a rat model of closed traumatic brain injury

BACKGROUND： Studies have reported the combined use of two-dimensional gel electrophoresis and mass spectrometry to detect differentially expressed proteins in the rat brainstem following brain injury. However, the detected differential proteins often exhibit low sensitivity and high relative molecular weight. Although protein chip technology is thought to compensate for these inadequacies, no related studies or results have been reported. OBJECTIVE： To propose the application of weak cation exchange protein chips in combination with mass spectrometry for determining protein expression profiles and characteristics in the brainstem following closed brain injury. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiments utilizing proteomics were performed from June 2007 to December 2008 in the Proteomics Laboratory, Medical College of Chinese People＇s Armed Police Force. MATERIALS： Weak cation exchange 2 protein chip, Ciphergen Proteinchip System （PBS-IIC）. METHODS： A total of 72 rats were randomly assigned to two groups： sham-surgery （n = 12） and injury （n = 60）. A closed traumatic brain injury model caused by falling object was replicated in the injury group, which was then subdivided into five subgroups according to different time points after injury： 4, 8, 12, 24, and 48 hours, with 12 rats in each subgroup. In the sham-surgery group, only the skin was removed and the stainless steel pad was fixed to the skull. MAIN OUTCOME MEASURES： The brain injury rats were sacrificed at 4, 8, 12, 24, and 48 hours after injury, respectively, and the control rats were sacrificed at 24 hours. Pathological changes in the brainstem were determined using hematoxylin-eosin staining, and differential protein expression in the brainstem was detected using a weak cation exchange 2 protein chip and protein chip reader. RESULTS： In the sham-surgery group, cells appeared normal. However, in the brain injury group, some brainstem neurons exhibited pyknosis, with reduced numbers of Nissl bodies in the cytoplasm swollen cell bodies and nuclei, irregular staining in the cytoplasm, and decreased numbers of neurons. Results from weak cation exchange 2 protein chip detection demonstrated that, compared with the sham-surgery group, the expression profiles of 2 proteins were altered in the brainstem of the injury group. At 12, 24, and 48 hours after injury, expression was increased （P 〈 0.01 ）. The mass charge ratio （M/Z） of 7 862 differentially expressed proteins was greater in the sham-surgery group compared with 12 and 24 hours after injury （P 〈 0.05）. CONCLUSION： The combined method of weak cation exchange 2 protein chip and mass spectrometry detected differential protein expression in the brainstem following closed brain injury in the rats, which suggested that closed brain injury induced altered protein expression profiles in the brainstem.

Using the SELDI ProteinChip System to Detect Changes in Protein Expression in Vero Cells after Infection

人的疱疹单一的病毒(HSV-1 ) 1 引起美容，眼睛，并且 encephalitic 疾病并且与潜伏的感染和癌症被联系。这里，我们开发了由使用 SELDI 蛋白质薄片在在 vitro 有教养的 Vero 房间检测蛋白质表示的变化在蛋白质水平学习 HSV-1 感染的致病的一个工具。在有为 12， 24 或 48 h 的 HSV-1 和文化的感染以后，房间被收获并且 lysed。IMAC3 数组被用于 SELDI-TOF-MS 在感染前后检测 proteomic 差别。薄片检测了一系列差别表示蛋白质山峰。有趣地，在 16 912 Da 的山峰和 17 581 Da 与 ISG15 的分子的团精确相应，它可以在感染的过程期间参予抗病毒的活动。因此，我们获得了的结果能用作一个基础学习在病毒和它的主人之间的 HSV-1 和相互作用的致病。另外，他们能为 HSV-1 感染的治疗在新治疗学的目标的发现帮助。关键词 SELDI 蛋白质薄片 - Vero 房间 - HSV-1 - 蛋白质表达式 CLC 数字 R373 基础项目：中国(30540075 ) 的国家自然科学基础；

Application of surface-enhanced laser desorption/ionization time-of-flight-based serum proteomic array technique for the early diagnosis of prostate cancer

Aim： To identify the serum biomarkers of prostate cancer （PCa） by protein chip and bioinformatics. Methods： Serum samples from 83 PCa patients and 95 healthy men were taken from a mass screening in Changchun, China. Protein profiling was carried out using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF MS）. The data of spectra were analyzed using two bioinformatics tools. Results： Eighteen serum differential proteins were identified in the PCa group compared with the control group （P 〈 0.01）. There were four proteins at the higher serum level and 14 proteins at the lower serum level in the PCa group. A decision tree classification algorithm that used an eight-protein mass pattern was developed to correctly classify the samples. A sensitivity of 92.0 % and a specificity of 96.7 % for the study group were obtained by comparing the PCa and control groups. Conclusion： We identified new serum biomarkers of PCa. SELDI-TOF MS coupled with a decision tree classification algorithm will provide a highly accurate and innovative approach for the early diagnosis of PCa. （Asian J Androl 2006 Jan; 8： 45-51）

Serum cytokine levels in chronic hepatitis B patients receiving peginterferon alpha-2a therapy

BACKGROUND: The relationship between cytokines and responses to peginterferon α-2a treatment in chronic hepatitis B patients has not yet been fully elucidated. We analyzed the serum levels of interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, vascular endothelial growth factor, interferon-γ, tumor necrosis factor-α, monocyte chemotactic protein-1 (MCP1) and epidermal growth factor during the treatment with peginterferon α-2a. METHODS: Ninety-three serum samples from 20 chronic hepatitis B patients were collected before, during and after 48 weeks of peginterferon therapy and were assayed for 12 cytokines. The patients were categorized as either virologic responders (VRs) or non-responders (NRs) according to their HBV DNA levels taken at 6th month during treatment. The Evidence Investigator (Randox, Antrim, UK), a protein chip analyzer, was used to quantify cytokines. RESULTS: Among the 12 cytokines, the levels of MCP1 were increased and the levels of IL-4 were decreased during the treatment in VRs. However these cytokines were not significantly changed in NRs in the treatment phases. Area under the receiver operating characteristic curve (AUROC) value of HBV DNA measured before the treatment was 0.81 in predicting VRs, and that of the baseline MCP1 was 0.76. IL-6 levels at 3rd and 6th months during the treatment also showed AUROC values 0.85 and 0.78 respectively in predicting sustained VRs. CONCLUSION: Serum cytokine levels reflect the pathological differences of individual treatment phases and could also be useful in monitoring responses to peginterferon treatment in chronic hepatitis B patients.

Fabrication of a Retinal-protein Photoreceptor Array

A pixel-architecture film of retinal proteins was prepared by an approach combining chemical, physical and biological technologies. Oriented multilayers of purple membrane composed of bacteriorhodopsin （BR） and lipids were patterned on an array of gold electrode pixels. In order to improve stability and resolution, the gene engineer- ing technique was employed to make a mutant of the protein BR by replacing the 36th amino acid residue from as- partic acid to cysteine with a thiol end group ready to react with gold; electric sedimentation was used to guarantee the high probability of formation of the Au-S bond and meanwhile to orient BR; further chemical crosslinking was introduced among layers of purple membranes to significantly enhance photoelectrical signals while keeping high stability. The non-bound BR region was eventually washed out by detergent, and the remaining BR pixels were thus detergent resistant due to chemical crosslinking among BR layers and covalent binding between the multilayer and the substrate. The protein array was confirmed to keep photoelectrical activity.

Protein microarray:A key approach of proteomics

Proteins are vital parts of living organisms,for they consist of the main components of physiological metabolic pathways.The expression and modification status of proteins in cells are time-and space-dependent based upon functions as well as responses to stress.A subject aiming at understanding of protein status under certain physiological condition in a large scale,termed proteomics,is fundamentally important in life science.Although the proteomic investigation mainly relies on electrophoresis,liquid chromatography,and mass spectrometry,protein microarray is emerging as one of new techniques,and its power is being recognized in the field recently.Herein,we highlight progress in protein microarray,both its generation and application.

Proteomics Technologies and Challenges

Proteomics is the study of proteins and their interactions in a cell. With the completion of the Human Genome Project, the emphasis is shifting to the protein compliment of the human organism. Because proteome reflects more accurately on the dynamic state of a cell, tissue, or organism, much is expected from proteomics to yield better disease markers for diagnosis and therapy monitoring. The advent of proteomics technologies for global detection and quantitation of proteins creates new opportunities and challenges for those seeking to gain greater understanding of diseases. High-throughput proteomics technologies combining with advanced bioinformatics are extensively used to identify molecular signatures of diseases based on protein pathways and signaling cascades. Mass spectrometry plays a vital role in proteomics and has become an indispensable tool for molecular and cellular biology. While the potential is great, many challenges and issues remain to be solved, such as mining low abundant proteins and integration of proteomics with genomics and metabolomics data. Nevertheless, proteomics is the foundation for constructing and extracting useful knowledge to biomedical research. In this review, a snapshot of contemporary issues in proteomics technologies is discussed.