Cell Research

Osteoarthritis is a major cause of chronic pain, but almost no one knows the molecular mechanism and the mediation osteoarthritis related joint pain. Therefore, treatment still shortage and joint replacement usually is inevitable. Here, we use a surgical model mice, and caught the long-term development of the osteoarthritis longitudinal evaluation pain related behavior and concomitant change in the dominant dorsal root ganglion (DRG). We prove monocyte chemoattractant protein (MCP), 1 (the CCL2) and its high affinity receptor, chemokine (c-c theme) receptor 2 (CCR2), is the core of the development of pain associated with osteoarthritis of the knee. The instability of the medial meniscus, laboratory mice early onset secondary mechanical allodynia, maintain 16 weeks. MCP 1 and CCR2 messenger RNA, proteins, and signal activity is temporary raised in eight weeks after surgery DRG control. The results and the movement of the statement caused pain behavior, this is to keep 16 weeks. Mice lack of Ccr2 also developed mechanical allodynia, but it began to solve from eight weeks. Despite the severe pain and structural knee damage equal to the wild type mice, mice Ccr2 - null did not develop sports pain behavior in eight weeks. In the wild type mice, macrophage infiltration to eight weeks of DRG, this is through the surgery keep 16 weeks. In contrast, macrophage infiltration not observed Ccr2 - null mice. These observations suggest a key role MCP 1 / CCR2 ways to establish osteoarthritis pain.

Food allergy (FA) is frequent in 8% of children under 3 years old and about 2% of adults. Chemokine is involved in all kinds of allergies such as fa. The purpose of this study is to determine the CCL2, CCL5, CCL11 level in the fa cup. The study population cross-sectional study contains 63 patients suffering from the fa cup and 100 healthy controls. The concentration of the CCL2, CCL5, CCL11, IgE measurement using enzyme-linked immunosorbent assay (ELISA). Use Kathy Ι eosinophils number cell count + analyzer system model SCAREF system GmbH. Is considered to be an important difference, P < 0.05. The results show that the number of FA patients significantly increased than cycle without disease control peripheral eosinophil. The fa patients serum immunoglobulin e level is higher than the control. We also show that serum CCL2 and CCL11 was significant increase in the fa, but compared with patients who CCL5 not detectable control. The results of this study showed that two CCL11 CCL2, more elevated FA children with allergic reaction and urticaria than bronchial asthma and allergic dermatitis. These results also show that more level of increase and CCL11 CCL2 observation after consumption of milk and pistachios. Overall, the results suggest that serum CCL2 and CCL11 rise FA and these may be considered useful parameters in the diagnosis of the disorder. Still can design treatment on the basis of, through the application block chemokine expression antibody they overcome allergic patients with complications in the fa.

Allele expression imbalance (AEI) is a kind of important genetic factor is the difference of the phenotypic traits, can genetic. The study can be used to find factors that regulate gene expression molecular mechanism, which is helpful to understand the bottom phenotypic change. Although it is generally accepted in many species, and we know that many genes show allele expression imbalance, this kind of phenomenon is not large scale study in cattle. Use pyrophosphoric acid sequencing technology method we analyzed a group of 29 cattle genes in order to find the favorable allele expression. The study investigated in three organization: liver, pituitary and kindey. From the research group of genes and their mineral wax (leptin), IGF2 (insulin-like growth factor 2), the CCL2 (chemokine carbon-carbon bond pattern ligand 2) display allelic expression is not balanced.

Alpha thrombin of serine protease is a kind of protein, can be used as the blood coagulation cascade. Thrombin procoagulant itself is a differences, fibrinogen make fibrin clot, but thrombin - thrombomodulin (TM) complicated start anticoagulation path, through the crack protein c. TM fragment of only the fifth and sixth egf like domain (TM56) enough to bind thrombin, but there are also the fourth egf like domain (TM456) induced vital anticoagulant activity of thrombin. Crystallography of thrombin - TM456 complex and not found in the thrombin bring significant structural change, this shows that TM4 may only provide a scaffold best calibration of protein C for the cleavage, thrombin. However, all kinds of experimental data show that exist TM4 may affect the dynamic properties of the active site cycle. In the present work, we use two traditional and accelerate the molecular dynamics simulation method is used to study the dynamic characteristics of the structure, thrombin of thrombin: TM56, thrombin: TM456 in a wide range of time scales. Two different but related allosteric way, while determining mediation pro - and anticoagulant blood clotting enzyme activity. A allosteric way, at the same time, present in the thrombin: TM56 and blood coagulation enzyme: TM456, direct connection TM5 domain of thrombin active site. Other allosteric path, which only appears in the slow time scale exist TM4 domain, including prolong the network related action connection TM4 and TM5 domain and active site cycle of thrombin.

Traumatic hemorrhage is stopped by a hemostatic plug formation, is from a complicated nonlinear and space heterogeneous network proteolytic enzyme biochemical called blood coagulation. We study the spatial dynamic thrombin, the enzyme this network, by developing a fluorogenic substrate to establish methods and space to solve imaging time, thrombin enzyme activity. Clotting stimulation, immobilized tissue factor induced localized clotting enzyme activities of the impulse, in space propagation, have all the characteristics of a tourist excitation wave: constant space velocity, constant amplitude and ignore the initial stimulus once more than activation threshold value. This wave parameters by phospholipid availability or platelet, wave not form in the plasma from type A hemophiliac factors or C lack of viii and XI, the two main mediators condensation positive feedback. Stimulation of the negative feedback of protein C ways and thrombomodulin produce nonstationary model of wave form followed by reduction and destruction. This suggests that the blood can be used as an emotional media, in tourism wave after wave of solidification.

Build a sensitive electrochemical aptamer sensor (aptasensor) to thrombin detection is described. In the use of the advantages of the equipment based on micro electrode the possibility of work with small sample capacity and enjoy faster mass transit interest rates and low interfacial capacitance than in macroelectrodes. Therefore, gold disk microelectrode array is an attractive sensor selection aptasensors. The morphology of the gold disk array microelectrode is through scanning electron microscopy. The interaction between, thrombin of clotting aptamer in gold disk microelectrode array is differential pulse voltammetry proved that using methylene blue (MB) as a kind of electrochemical indicator. MB adsorption to the aptamers through their guanine base. Introduces the thrombin, it replaced the MB adsorption to the aptamers and bound to them. This leads to reduce MB peak current is also related to concentration of thrombin in a dynamic range across from 10 (5) to 10 (- 12) M. This method can selectively detection thrombin linear and detection limit for 0.143 points.

We previously proposal, epithelial cell adhesion molecule positive (EpCAM (+)) of hepatocellular carcinoma (HCC) cells form a population of initiating the tumorigenic cells and liver cell carcinoma specimens line. Here, EpCAM (+) circulating tumor cells (CTC) in liver cell carcinoma patients prospectively determine the effect and prognosis of significanceand resection, their stem cell characteristics for further investigation. From 123 patients with hepatocellular carcinoma of the blood sample testing and 1 month ago after resection.

This study is to evaluate the relationship between the organization and the patient's prognosis BMP7 expression in hepatocellular carcinoma (HCC). BMP7 mRNA expression in liver cell carcinoma characteristics is the use of real time PCR and 30 double fresh frozen liver cell carcinoma tissues and the corresponding not cancerous tissue. BMP7 protein expression in liver cell carcinoma is confirmed using immunohistochemistry in tissue microarray. Finally, BMP7 expression and routine clinical pathology characteristic and the prognosis of patients with hepatocellular carcinoma (HCC). BMP7 mRNA expression and protein in liver cancer cell is far higher than normal liver cells. Our results show that high expression BMP7 in liver cell carcinoma on tumor size (p < 0.001), tumor cell differentiation (p = 0.041), serum AFP (p = 0.007), and tumor stage (p < 0.001). Kaplan meier survival analysis showed that a high expression levels of BMP7 lead to obvious liver cell carcinoma patients prognosis. Multivariate analysis showed that BMP7 expression level is an independent prognostic parameters on the overall survival rate for patients with hepatocellular carcinoma. These findings provide evidence shows that a high expression levels of BMP7 as a specific biomarker for the prognosis of patients with hepatocellular carcinoma (HCC) bad. Therefore, we speculate that may be a potential target BMP7 antiangiogenic therapy of liver cancer.

Using fluorescence in situ hybridization (FISH) analysis, we examined the replication mode of the centromere region (homologous counterpart) and the aneuploidy level of chromosome 17 in the interphase nuclei of phytohaemagglutinin (PHA)-stimulated peripheral blood lymphocytes from (1) patients with hepatocellular carcinoma (HCC); (2) patients with liver cirrhosis (LC) due to hepatitis C viral infection who are individuals at a higher increased risk for HCC; and (3) healthy control participants. We also compared the allelic-replication asynchrony and aneuploidy frequencies with serum alpha-fetoprotein (AFP) levels. We found a significant increase in centromeric replication asynchrony accompanied by a high frequency of aneuploidy in lymphocytes of HCC patients compared with those of LC patients and healthy control participants. These changes are similar to those previously observed in other types of malignancy (hematological, ovarian, prostate, and breast cancer). The cytogenetic alterations of aneuploidy and strong asynchronous replication displayed in the lymphocytes of HCC patients arose from malignancy, as they were associated neither with an increased risk for cancer nor with an infection. The cytogenetic cancer-associated markers observed in patients' lymphocytes appeared to be superior to serum AFP, the marker currently used for HCC. Thus, the cytogenetic cancer-associated markers may be potentially useful in noninvasive cancer detection

Secretion clusterin (sClu) is a kind of disease of protein, played an important role in protecting cells from tumor necrosis factor (TNF) α induced apoptosis. The purpose of this study was to survey the molecular mechanism of the influence sClu TNF induced apoptosis in breast cancer cells. The wild type of p53 expression MCF 7 cell line was designed out excessive performance sClu (MCF 7 / sClu), and mda - m - 231 cell line and mutant p53 and sClu son transfection is the silence of the small interference RNA (mda - m - 231 / sClu small interference RNA). Clusterin overexpression influence, in the cell apoptosis and sensitivity are two TNF α in vitro. Our results show that TNF treatment increase BCL 2 mRNA and protein content in breast cancer cells, which suggests that BCL 2 directly subject to nuclear factor - kappaB (NF - kappaB) in response to TNF α. The induction of the base class library 2 mediated p65 subelement of NF - kappaB. The silence of the small interference RNA mediated BCL 2 lead to increased significantly TNF induced apoptosis. The silence of the sClu in mda - m - 231 / sClu small interference RNA cell abolished TNF mediated NF - kappaB activation and BCL 2 overexpression, and present mda - m - 231 / sClu small interference RNA cell significantly more sensitive, TNF than parent mediated apoptosis. In addition, sClu expression in the MCF 7 / sClu cell mediated TNF NF - kappaB promotion activities and BCL 2 overexpression, and presents MCF 7 / Clu cell significant resistance to TNF α mediated apoptosis. Inhibition of NF - kappaB activity or p65 and BCL 2 expression reverse these effects. The results show that sClu awarded breast cancer cells apoptosis induced resistance of TNF α by NF - kappaB activation and BCL 2 overexpression.

The skin is often damaged by genetic and environmental factors such as smoking, contact xenobiotics, heat, hormonal changes, and ultraviolet ray. These factors will cause skin disease. White rattan is quadrangularis falls. (CQ) had been used in folk medicine to treat skin disease since ancient times. In considering take medicinal values show this genus, this is decided to investigate the antitumor activity of CQ. Extraction get and their phenolic content by in vitro CQ antitumor activity evaluation by using A431 (skin epidermoid carcinoma, humans) cell line. A431 cells are used different extract of CQ in dose dependent way. Five extract, acetone extract demonstrates important anticancer activity in A431 cell line. N-hexane, chloroform, ethyl acetate and methanol extract also showed a cell toxicity, but for a relatively small extent than acetone extract. GI (50) value of the acetone extract was found to 8 μ g ml (1), and GI (50) value of the purification part of acetone extract, known as the AFCQ (active part acetone CQ) A431 cells, were found to be 4.8 u g ml (1). In addition, the mechanism of antitumor activity, AFCQ exhibited by comparing its effect and the standard anti-cancer drugs adriamycin (doxorubicin) through the evaluation of the state mark apoptosis after treatment A431 cells and AFCQ and adriamycin. Burlington BCL 2 ratio along with the release of cytochrome c from the mitochondrial cytoplasm, this is a hallmark of cell apoptosis, also be evaluated. Cleavage of PARP revealed that AFCQ A431 cells apoptosis occurred, reference doxorubicin.

Antisense oligonucleotides (management) has been used for prostate cancer model for growth regulatory proteins, and at least one oligosaccharides (against BCL 2) has reached the clinical test. We've found that in LNCaP cell, mono - and double function management, the relative inhibitory expression, as compensation, the base class library 2 inhibition caspase 3 (apoptosis promoter) activities, and increase the expression of androgen receptor (AR) and p300 and il - co catalyst. In addition, the prostate specific membrane antigen (PSMA) and (may be its regulators) interferon (IFN) rise. 14 protein distributed between regulators of apoptosis, androgen control, differentiation antigen and autocrine mediated growth has been inspection. We put these findings including vascular endothelial growth factor (VEGF), promote angiogenesis, not through the compensation significant change, so do not need additional supervision for repression BCL 2 treatment is effective (such as caspase 3).

Invertebrate lysozyme (I - lyz or destabilase) exist in shrimp. The protein may have a function as a peptidoglycan break enzyme and as a peptide enzyme. Shrimp usually vibrio infection. , gram negative bacteria, as is known to all, c - lyz (similar to the chicken lysozyme) is active against these bacteria. In order to further understand renaturation of rules, we determine the sequences of genes and protein structure I - lyz modeling. In addition, I - lyz expression and c - lyz response lipopolysaccharide (LPS) was studied. Shrimp I - lyz gene is made up of two introns interrupt to standardize joint connection. Expression of shrimp I - lyz is transient cut injection followed by after LPS induced after 6 hours in hepatopancreas. By contrast, c - lyz is raised hepatopancreas 4 h post injection and trimmed in gill. L. vannamei I - lyz does not contain of lysozyme catalytic residues (glycohydrolase) is not isopeptidase activities; However, as is known to all, antibacterial activities not only rely on the enzyme activity of protein. Research invertebrates lysozyme will increase our understanding of the regulatory process defense mechanism.

The study describes the use of selective high PI protein extraction and enrichment of human serum albumin (HSA) modified gold nanoparticles (HSA-AuNPs), lysozyme (LYZ) prior to analysis by capillary electrophoresis (CE)with UV detection. HSA gold nanoparticles can be extracted from a complex matrix LYZ HSA the caprock not only stable gold nanoparticles in high salt environment, but also showed a strong electrostatic attraction LYZ neutral pH conditions. LYZ and other high isoelectric point (pI) of the efficient separation of the protein has been successfully achieved by the cationic polyelectrolyte, poly (dimethyl diallyl ammonium chloride the) (PDDAC) filling, background electrolyte. Capture and HSA-AuNPs LYZ PDDAC filled CE can be directly used for analysis of the extracted LYZ releasing agent is not added to the extractor. Extraction efficiency depends on the gold nanoparticles of the solution pH and concentration of HSA. In the preferred extraction conditions, the signal-to-noise ratio of the detection limit of 3 LYZ of 8nm or less. HSA-AuNPs extraction and PDDAC of a combination of filling the CE has been the application of the analysis LYZ egg white, human milk, and the human tear. In addition, based on the extraction of the NP may be coupled to matrix-assisted desorption / ionization time-of-flight mass spectrometry and sodium dodecyl sulfate - polyacrylamide gel electrophoresis

At the transcriptional level lysozyme (LYZ), surf clams clams the four corners clams of defense components combined exposure to heavy metals and bacterial reaction metallothionein (MT) and superoxide dismutase (SOD) were assessed. First, the length LYZ cDNA containing 808 nucleotides and coding for 194 deduced amino acid sequence is determined from the clam. The multiple alignment a discovery, MvLYZ high recognition and invertebrates LYZs from other mollusks. Next, clams exposed to Vibrio parahaemolyticus and a mixture of cadmium and mercury, singly or in combination, the seven days. Part of three lines of defense of the cumulative mortality of clams and mRNA expression analysis. The highest mortality occurred in the first seven days of comprehensive treatment. Three gene expression is upregulated in response, compared with the control of the different response time, and the transcription level of treatment; just occurred in response to the combined exposure, rather than a single exposure. Experimental group, MvLYZ the expression and expression of MvSOD reached its peak, in the first three days of comprehensive treatment, MvMT expression peaked in the first five days of heavy metal treatment. In addition, the interactive effects of heavy metals and vice transcription reaction changing the exposure time. Therefore, the joint exposure under the transcriptional regulation of the three genes from a single exposure is much more complicated.

Numerous sequence-specific transcription factors (transcription factors, TFs) a combination of cross-regulatory network, these regulatory networks constitute the basis of the cells play a biological function. On September 5, the internationally renowned journal Cell Online published an article entitled Circuitry and Dynamics of the Washington University researchers transcription factor regulation network loop dynamics and organization of Human Transcription Factor Regulatory Networks research papers, reports principles. The researchers used genome in vivo DNaseI footprinting, 475 sequence-specific transcription factor found in 41 different cells and tissues, and then analyze the dynamics of these linkages. The study found that the transcription factor network with a high degree of cell selectivity, cell selectivity is driven by several factors, including in the control of cell identity unrecognized regulon. And, also identified a variety of widely expressed factors affect the transcriptional regulatory network. Shocking, the Although transcription factors inherent diversity, independent regulatory networks of all cell types converge with neural network like life. This study provides extensive description of the transcription factor regulatory network loop dynamics and organizational principles.

FGF-21 biological functions mainly as the metabolic regulation hungry, but also start a major regulator of ketone body production factor. Studies have shown that patients with a variety of diseases, including diabetes, blood levels of FGF-21 significantly increased. Long-term fasting induced mouse liver expression of FGF-21 protein by peroxisome proliferator-activated receptor (PPAR) alpha pathway significantly with peripheral FGF-21 levels, and thus promote adipose tissue lipolysis and fatty acid to The outer periphery of release of fatty acids in the liver and then absorbed and converted to ketones. Another study shows that the role of FGF21 fibroblast growth factor 21 may be assisted by inducing PPAR-γ activation factor (PGC) 1α to complete, which can stimulate liver gluconeogenesis, fatty acid oxidation and ketone body production. Recent studies have shown that, FGF-21 can induce liver secretes pancreatic lipase. Liver pancreatic lipase will be a substantial increase in the metabolic state of low energy, so the FGF-21 that may be involved in the regulation of the metabolic state of low energy. FGF-21 transgenic mice were fasted for 24 hours into the metabolic state of the low-energy, generate large amounts of ketones, and normal mice do not. In addition, FGF21 fibroblast growth factor 21 transgenic mice petite, weighing only 40% to 50% of the normal mice. Hunger mainly by the growth hormone / insulin-like growth factor 1 (GH/IGF-1) way to inhibit the growth and FGF-21 can be activated by reducing the signal transduction and transcription factor (STAT) 5 activation to significantly reduce serum IGF content Therefore, FGF-21 in the state of hunger by inhibiting the growth to save energy. Thiazole TZDs (TZD) can induce db / db mice white fat and 3T3-L1 fat cells FGF-21 The expression. Rosiglitazone and pioglitazone increase insulin sensitivity through PPAR-γ, and white adipose PPAR-γ can induce FGF-21 expression, therefore TZD FGF-21 may play a role. Interestingly, the existence of circadian rhythm: FGF-21 The expression of PPAR-α ligands injected mice night bezafibrate can significantly induced the FGF-21 expression, while the daytime injection does not appear to the performance. Meanwhile, such a regulatory role in the PPAR-α knockout mice did not show that induced the expression of FGF-21 circadian bezafibrate is effected by circadian reactive liver PPAR-α system to determine. Human FGF-21 has been reported. Fasted for 7 days after the blood content of FGF-21 increases, and in patients with hypertriglyceridemia vivo FGF21 fibroblast growth factor 21 content 2 times higher than normal, and will further increase after taking fenofibrate. These data show that fasting can induce the expression of the human FGF-21, which may be regulated by PPAR-α. Other studies have shown that obesity and type 2 diabetes in patients with FGF-21 levels were significantly increased, and fat, insulin and triglyceride levels were positively correlated, but negatively correlated with high density lipoprotein. In addition, the FGF-21 mRNA expression in subcutaneous fat and blood content of FGF-21 significantly correlated prompted adipose tissue may be one of the main source of the blood FGF-21. Recent studies show that non-alcoholic fatty liver disease, chronic kidney disease and coronary heart disease in patients with blood FGF-21 levels were also significantly elevated.

Human dendritic cells originated from hematopoietic stem cells (hemopoieticstemcell). DC source of two ways: ① myeloid stem cells in the GM-CSF stimulated to differentiate into DC called myeloid DC (myeloid dendritic cells, the MDC), also known DCl, monocytes and granulocytes have a common precursor cells; Langerhans cells (Langerhans cells, LCs), the mesothelial (or dermal) DCs, and monocyte-derived DCs ② derived from lymphoid stem cells, known as lymphoid the DC (Lymophiod dendritic cells, LDC ) or plasmacytoid DC (plasmacytoid dendritic cells, pIX ;), i.e., DC2, and a common precursor cells of T cells and NK cells. Although the number of dendritic cells (DC) less than 1% of peripheral blood mononuclear cells, but the surface has a rich antigen presenting molecules (MHC-Ⅰ and MHC-II), co-stimulatory factor the (CD80/B7-1 CD86 / B7-2, CD40, CD40L, etc.) and adhesion factor (ICAM-1, ICAM-2, ICAM-3, LFA-1, LFA-3, etc.), is a powerful professional antigen presenting cells (APC). DC has its own the immunostimulatory ability, found the only way to activate naive T cells are not sensitized APC.

The bone marrow mesenchymal stem cells (mesenchymal stem cells, MSC) is an important member of the family stem cells, derived from the development of the early mesoderm and ectoderm. MSC was originally found in the bone marrow, increasing people's attention because it has multiple differentiation potential, hematopoietic support and promote the implantation of stem cells, immune regulation and self-replication. Such as mesenchymal stem cells under the particular inducing conditions in vivo or in vitro, can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac, endothelial and other tissue cells, continuous passage culture and cryopreservation still has multiple differentiation potential, can be used as an ideal seed cells for aging and lesions caused by tissue and organ damage repair. Bone marrow mesenchymal stem cells because of its wide range of sources, easy isolation and culture, and has strong differentiation potential and advantages of autologous transplantation, more and more scholars of all ages, is considered to be soon be introduced optimal clinical treatment stem cells . Mesenchymal stem cells in the bone marrow has the following advantages: Have a strong proliferation capability and multilineage differentiation potential, not only to differentiate into hematopoietic cells in a suitable in vivo or in vitro environments, and also having the differentiation of muscle cells, liver cells, osteoblasts, chondrocytes, and stromal cells variety the ability of the cells. Two immunomodulatory functions through the interaction between the cells and the to produce cytokines inhibit T cell proliferation and immune response, thus play the function of the immune reconstitution. Three. Has a convenient source of easy separation, culture, amplification and purification the multiple passages amplification still has the characteristics of stem cells, there is no immune rejection characteristics.

According to the American physicist organizational networks reported on June 29, U.S. scientists found cytokines SAMHD1 protein inhibition mechanism of bone marrow cells infected with HIV (the AIDS virus), new research extends the people how to evade the immune system of AIDS patients how to deal with HIV and HIV the understanding of the immune response in order to find to prevent HIV infection or to prevent the new therapies for its replication in the infected individuals. The research is published in the June 30 issue of Nature magazine. Bone marrow cells are the antigen a subset of the white blood cells, the immune response against viruses and other pathogens is very important for the body. Jack Scott the Jaworski's research team at Case Western Reserve University School of Medicine, AIDS Research Center and the Stowers Institute for Medical Research in the United States, Michael Walsh led a team SAMHD1 protein that senses such as macrophages and dendritic cells and bone marrow cells infected with HIV-1 virus (HIV divided into type 1 and type 2, type 1 is the main strains of the global epidemic of type 2 only popular in West Africa) and other immunodeficiency virus copies of the virus and prevent synthesis in these cells, thereby inhibiting the HIV virus. Previously, scientists have always thought that, SAMHD1 mutation can cause symptoms called AGS and symptoms of congenital infection, to be attributed to the absence of the virus, the immune system interferon inappropriate induction. Therefore, SAMHD1 and other the initiator the AGS cell proteins can abandon the nucleic acid fragments of the cells, and the prevention of such interferon system is activated inappropriately.