The study on using dispase and trypsine 0.25% to detach cells from fetal monkey kidneys of Macaca mulatta showed that: the detach ability of dispase was much higher than trypsin. Some characteristics of dispase were suitable for detaching fetal cell in cold condition or serum condition. However, dispase has not been used as commonly as trypsin. Dispase can not be a good solution to detach cells without the effect of trypsin
Endopeptidase ; cells ; Kidney ; Haplorhini ; Fetus

In several human tumors, signal transducer and activator of transcription 3 (STAT3) and nuclear factor κB (NFκB) are activated and interact; how these STAT3–NFκB complexes are transported to the nucleus is not fully understood. In this study, we found that Rac1 was activated in starved cancer cells and that activated Rac1 coexisted with STAT3 and NFκB. Rac1 knockdown and overexpression of the dominant-negative mutant Rac1N19 inhibited the degradation of IκBα, an inhibitor of NFκB. MG132, an inhibitor of the ubiquitin proteasome pathway, increased the amount of non-phosphorylated IκBα, but not serine-phosphorylated IκBα, indicating that IκBα degradation by Rac1 in starved cancer cells is independent of IκBα serine phosphorylation by IKK. Rac1 knockdown also inhibited the nuclear translocation of STAT3–NFκB complexes, indicating that this translocation requires activated Rac1. We also demonstrated that the mutant STAT3 Y705F could form complexes with NFκB, and these unphosphorylated STAT3–NFκB complexes translocated into the nucleus and upregulated the activity of NFκB in starved cancer cells, suggesting that phosphorylation of STAT3 is not essential for its translocation. To our knowledge, this is the first study demonstrating the crucial role of Rac1 in the function of STAT3–NFκB complexes in starved cancer cells and implies that targeting Rac1 may have future therapeutic significance in cancer therapy.
Humans ; Phosphorylation ; Proteasome Endopeptidase Complex ; Serine ; STAT3 Transcription Factor ; Ubiquitin

It has been suggested that proteasome system has a role in initiation, progression, and complication stages of atherosclerosis. Although there is still controversy, there has been no research that compares the expression of proteasome in tissue and serum at each of these stages. This study aimed to investigated the expression of proteasome at different stages of atherosclerosis using rat model. We measured the expression of aortic proteasome by immunohistochemical analyses and were then analyzed using ImageJ software for percentage of area and integrated density. We used Photoshop version 3.0 to analyze aortic proteasome expression as a comparison. We measured serum proteasome expression by enzyme linked immunosorbents assays. Kruskal-Wallis test was used to compare mean value of percentage of area and serum proteasome. Analysis of variance test was used to compare mean value of integrated density. Correlation test between vascular proteasome expression and serum proteasome expression was made using Spearman test. A P-value of 0.05 was considered statistically significant. Compared with normal, percentage of area was higher in initiation, progression, and complication. Compared with normal, integrated density was higher in initiation and further higher in progression and complication. Data from Image J is similar with data from Photoshop. Serum proteasome expression was higher in initiation compared with normal, and further higher in progression and complication. It was concluded that there were different vascular proteasome expression and serum proteasome expression at the stages of atherosclerosis. These results may be used in research into new marker and therapeutic target in atherosclerosis.
Animals ; Atherosclerosis* ; Immunosorbents ; Models, Animal ; Proteasome Endopeptidase Complex* ; Rats*

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex

Small RNAs, especially microRNAs (miRNAs),widely exist in eukaryotic cells, with their main functions being regulating gene expression and function of target molecules through the degradation of cellular target RNAs by the ribonuclease-based system. Ubiquitins and ubiquitin-like proteins are polypeptides that exist in most eukaryotic cells, and their main function is almost to regulate protein level through the degradation of cellular proteins by ubiquitin proteasome system. Small RNAs, including miRNAs,and ubiquitins or ubiquitin-like proteins have similarities in many aspects although small RNAs and ubiquitin or ubiquitin-like proteins interact different substrates respectively. Therefore, miRNAs can be defined as ubiquitra (ubiquitous ribonucleic acid, ubiquitra or uRNA), and the other small RNAs can be defined as ubiquitra-like RNA or uRNA-like RNA. The concept of ubiquitra may be applied for explaining the biological essence of small RNAs diversity.
Gene Expression ; MicroRNAs ; Proteasome Endopeptidase Complex ; Proteins ; Ubiquitination

BACKGROUND: Proteasomes are multi-subunit enzyme complexes present in the cytoplasm and nucleus of eukaryotic cells. Proteasomes are involved in the pathophysiological process resulting in development of many diseases. Release of proteasomes from lyzed erythrocytes has been suggested in recent reports. Accumulation of proteasomes in blood products could contribute to formation of storage lesions and have adverse effects on recipients; therefore, we conducted an analysis of changes in concentration of proteasomes in blood products during storage. METHODS: Concentrations of 20S proteasomes in supernatant of whole blood products obtained from eight healthy volunteers and in segments of 16 packed red blood cell (pRBC) units transfused to patients were measured by ELISA. Plasma samples containing several hemoglobin concentrations were prepared in order to assess the relationship between proteasome concentration and degree of hemolysis. RESULTS: Proteasome concentrations in whole blood products on day one of storage were significantly lower than those on day seven of storage and later (P<0.05). In segments of pRBC units, the proteasome concentration was 8.072+/-11.802 microg/mL (storage day: 13.8+/-4.7). Of the 32 pRBC units, two showed extremely high proteasome concentrations (36.662 and 62.798 microg/mL). Proteasome concentrations in plasma increased with increasing hemoglobin concentrations. CONCLUSION: During storage of whole blood products, except during the first seven storage days, levels of proteasome do not undergo significant change. However, hemolysis may be related to accumulation of proteasome. Further study to evaluate the effects of blood components containing high proteasome concentrations on recipients should be conducted.
Cytoplasm ; Enzyme-Linked Immunosorbent Assay ; Erythrocytes ; Eukaryotic Cells ; Hemoglobins ; Hemolysis ; Humans ; Plasma ; Proteasome Endopeptidase Complex

Antibody-mediated rejection (AMR) by preformed and/or de novo human leukocyte antigen alloantibodies is a leading cause of early and late allograft loss. In this review, we describe strategic approaches to various forms of AMR in clinical settings that are not based on pathologic classification, which is controversial for atypical AMR (C4d-, DSA-, subclinical etc.). For acute AMR, a variety of modalities like plasmapheresis, intravenous immunoglobulin, and anti-CD20 antibodies have been utilized singly, or in combination, with variable results; however, no established treatment for chronic AMR is known. Significant research efforts are being made for developing new and novel therapies. Improvements in clinical outcomes can be expected from studies evaluating innovative therapeutic concepts, such as proteasome inhibition or complement-blocking agents.
Antibodies ; Humans ; Immunoglobulins ; Isoantibodies ; Leukocytes ; Plasmapheresis ; Proteasome Endopeptidase Complex ; Rejection (Psychology) ; Transplantation, Homologous

Orientia tsutsugamushi, the causative agent of scrub typhus, is an obligate intracellular bacterium that replicates in the cytosol of host cells. Although several protein antigens have been characterized and cloned, little information exists regarding the polysaccharide antigen of this bacterium. In this study, we characterized two monoclonal antibodies, NT19 and WT14, against the proteinase K-resistant antigen of O. tsutsugamushi. Western blot analysis showed that MAb NT19 and WT14 strongly recognized two antigenic bands with molecular masses of 20 kDa and 24 kDa, which were resistant to proteinase K digestion. We suggest that the proteinase-resistant antigen might be polysaccharide. One patient serum reacted with a 24 kDa band that was similar to a band observed by WT14, suggesting the possibility of the role of this proteinase-resistant antigen as an antigenic molecule in human infection.
Antibodies, Monoclonal ; Blotting, Western ; Clone Cells ; Cytosol ; Digestion ; Endopeptidase K ; Humans ; Orientia tsutsugamushi ; Scrub Typhus

Drowning is one of the most common causes accidental death worldwide, but its diagnosis remains a challenging task in forensic pathology. Several authors have suggested that diatom analysis be conducted via an enzymatic digestion method that uses proteinase K to provide objective evidence for drowning; we employed this method in our study because of its superior applicability as compared to the conventional disorganization methods. The purpose of this study was to examine the reclaiming ratio of diatoms from experimentally drowned animal organs, which could be influenced by diatom morphology. The authors injected 3 diatoms species (Cyclotella striata, Navicula incerta, and Pleurosigma angulatum) into a rat's airway and compared the detection rate to investigate the factors that influence the sensitivity of diatom analysis. The results are as follows: (1) Average reclaiming ratio in the lungs was 81.07 for Navicula incerta, 48.26 for Cyclotella striata, and 5.35 for Pleurosigma angulatum. (2) The detection rates from the closed organs in 15 experimental animals were highest in the kidney (73%, 11/15), followed by the heart (67%, 10/15), brain (60%, 9/15), and liver (53%, 8/15). (3) Two Cyclotella striata was detected in the kidney of postmortem control group which suggest the possibility of contamination during laboratory procedure. In conclusion, the authors propose that diatom size could be a significant influencing factor for diatom extraction from the organs of drowned bodies; therefore, the results of diatom analysis must be interpreted after considering the diatom population of the drowning medium at the scene and the possibility of contamination during the laboratory procedure.
Animal Structures ; Animals ; Brain ; Diatoms ; Digestion ; Drowning ; Endopeptidase K ; Forensic Pathology ; Heart ; Kidney ; Liver ; Lung

The fibrillar coat of Candida albicans is of interest as its significance in antigenicity, antiphagocytosis, and adherence to host tissues. The partial biochemical properties and ultrastructure of fibrillar coat induced by rabbit sera were examined. The induced fibrillar layer was destroyed by treatments of lyticase, proteinase K and dithiothreitol. The total protein concentration of fibrillar cell wall lysate was higher than that of non-fibrillar cell wall lysate, but the total sugar concentration was similar. On SDS-PAGE analysis, the protein profiles between in fibrillar cells and in non-fibrillar cells were shown to be different. In fibrillar cells, the major bands of cell wall lysate were 83, 66, 54, 47, 33, and 26 kDa in dithiothreitol-treated lysate. The proteins of 26 and 19 kDa were predominant in lyticase-treated lysate. Although the fibrillar thickness and protein amount of cell wall lysate were increased in according to the incubation time, the protein profiles did not changed. These results suggest that the proteins of 83, 66, 54, 47, 33, 26, and 19 kDa may be major constituents of fibrillar coat in C. albicans.
Candida albicans* ; Candida* ; Cell Wall ; Dithiothreitol ; Electrophoresis, Polyacrylamide Gel ; Endopeptidase K