As the major pathway mediating specific protein degradation in eukaryotes, ubiquitin-proteasome system (UPS) is involved in various physiological and pathological processes such as cell cycle regulation, immune response, signal transduction and DNA-repair. Deubiquitinases (DUB) maintain the balance of UPS and related physiological processes via reversibly removing ubiquitin from the covalently modified protein substrates, which have been implicated in various disease processes in case of their imbalance expression. Because DUB plays critical regulating roles in the UPS pathway, they may be also the ideal drug targets for severe and intractable human diseases, such as cancer and neurodegenerative disease. With the rapid development of proteomic technology, systematical investigation of specific substrates and interacting proteins of varied DUB via mass spectrometry approach may shed light on these DUB's biological function and regulating roles in the physiological and pathogenic states. In this review, we briefly introduce the characteristics of DUB and summarize the recent application and progresses of proteomics in DUB research.
Humans ; Mass Spectrometry ; Proteasome Endopeptidase Complex ; metabolism ; Proteomics ; Signal Transduction ; Ubiquitin ; metabolism ; Ubiquitin-Specific Proteases ; metabolism

Protein ubiquitination is one of the most important and widely exist protein post-translational modifications in eukaryotic cells, which takes the ubiquitin and ubiquitin chains as signal molecules to covalently modify other protein substrates. It plays an important roles in the control of almost all of the life processes, including gene transcription and translation, signal transduction and cell-cycle progression, besides classical 26S protesome degradation pathway. Varied modification sites in the same substrates as well as different types of ubiquitin linkages in the same modification sites contain different structural information, which conduct different signal or even determine the fate of the protein substrates in the cell. Any abnormalities in ubiquitin chain formation or its modification process may cause severe problem in maintaining the balance of intracellular environment and finally result in serious health problem of human being. In this review, we discussed the discovery, genetic characteristics and the crystal structure of the ubiquitin. We also emphasized the recent progresses of the assembly processes, structure and their biological function of ubiquitin chains. The relationship between the disregulation and related human diseases has also been discussed. These progress will shed light on the complexity of proteome, which may also provide tools in the new drug research and development processes.
Humans ; Proteome ; Ubiquitin ; chemistry ; Ubiquitination

Objective To analyze the relationship between blood glucose level, blood pressure and weight with pancreatic cancer genesis. Then to explore the metabolism associated risk factors in pancreatic cancer genesis. Methods Form December 2002 to September 2009 in Ruijin Hospital, 548 pancreatic cancers with pathology diagnosis after pancreatectomy were collected for the study with retrospective analysis method. The association of pancreatic cancer with blood glucose level, blood pressure, weight and other metabolic factors were analyzed. Results With principal component analysis, it suggested that there were strong correlation between blood glucose level, blood pressure and weight index (BMI) increasing with pancreatic cancer. The contribution rates were 3. 614%,25. 236%, 15. 418% and 12. 918%, respectively. Single factor analysis indicated that the association between pancreatic cancers and new onset diabetes mellitus (duration≤ 2 years) was stronger than that of long-term diabetes mellitus. The occurrence rate of pancreatic cancer in patients with long-term diabetes whose blood glucose level was not well controlled recently while well controlled previously (44.6 % ) was significant hister than that in patients without diabetes (5. 6% , P＜0.05). The fasting blood glucose level of these PC patients ( 13.87± 3. 49 mmol/L) was significantly higher than new onset and other long-term diabetes patients, the comparative risk was 13.46 (95% CI 4. 560,39. 731). BMI increasing was a risk factor of pancreatic cancer, but there was no significant statistical difference between risk degree and BMI increasing level. All above metabolic diseases were risk factors of pancreatic cancer, but for pathology, location and stage of pancreatic cancer there was no statistical difference in theses factors. Conclusion This study suggested diabetes, BMI increasing and hypertension were high risk factors of pancreatic cancer genesis. New onset and long-term diabetes patients whose blood glucose not controlled well recently should be watched carefully for pancreatic cancer. Early treatment and intensive follow-up of metabolic disease might be helpful to early diagnosis and prognosis of pancreatic cancer.

Objective We increase the soluble expression of artificial tandem hybrid ubiquitin binding domains ( ThUBD) in prokaryotic cytoplasm of Escherichia coli BL21 ( DE3 ) , which offer an effective and special profiling for ubiquitin conjugates( UbC) .Methods Codon optimization of the ThUBD was performed, followed by analysis of codon relative adaptiveness based on relative frequency of synonymous codon ( RFSC) of E.coli.Further induced expression and yeast ubiquitin conjugate enrichment quantified the soluble ThUBD-S and tested the ability to bind UbC.Results The statistical result showed that the percentage of codon of the highest usage frequency was increased from 48%to 75%, and codon adaptation index( CAI) was increased from 0.63 to 0.88 after codon optimization, which might suggest a higher expression of the ThUBD in E.coli BL21 (DE3).The subsequent SDS-PAGE indicated that the soluble target protein was increased four times, which accounted for 13.06%of total cell lysis.Further ubiquitinated proteome of yeast demonstrated that the ability to bind and enrich UbC of optimized ThUBD-S did not change compared with original ThUBD.Conclusion The expression of ThUBD-S can quadruple after codon optimization.At the same time, codon optimization does not impact its soluble expression and the ability to bind UbC.

Objective To study the effect of siRNA targeting bcl-2 gene on the drug-sensitivity of HL-60 cell to homoharringtonin(HT). Methods siRNA, selected by previous experiments and packaged with lipofectamine 2000, was transferred into HL-60 cells. Six hours later, the cells were cultured with homoharringtonin. The cell growth of HL-60 cells was detected by MTT methods at 24, 48, 72 hours respectively. The level of bcl-2 protein and ROS (reactive oxygen species, ROS) as well as membrane potential of mitochondrion was determined by flowcytometry. Results The siRNA could significantly increase the inhibitive effect of homoharringtonin on growth of HL-60 cells. The combination of siRNA with homoharringtonin resulted in the falling of bcl-2 protein level and rising of ROS level as well as descending of membrane potential of mitochondrion of HL-60 significantly (P

OBJECTIVE To investigate the clinical features and causes of nosocomial infection in schizophrenics with tuberculosis so as to take effective measures for better prevention and treatment.METHODS A retrospective investigation was used to analyze the clinical features and causes of nosocomial infection in 207 schizophrenics with tuberculosis.RESULTS The rate of nosocomial infection in schizophrenics with tuberculosis was 39.13%.The main infection way was via respiratory tract,followed by skin,soft tissue and gastrointestinal tract.The period and degrees of tuberculosis,the history of smoking,psychiatric symptoms and the complications were the causes of nosocomial infection.CONCLUSIONS Controlling psychiatric symptoms actively,preventing and treating somatic complications,using antibiotics rationally and less or no smoking are the effective measures to avoid nosocomial infection in schizophrenics with tuberculosis.

Objective To study the clinical significance of P - selection expression in children with viral encephalitis and the correlation between this expression and the cerebral infarction with critical viral encephalitis. Methods Flow cytometric was employed to detect the expression of P- selection on the surface of platelet membrane in 44 children with viral encephalitis(20 light patients and 24 critical patients) and 20 healthy control children. The area of the cerebral infarction was determined by computed tomographic scan in 20 patients with critical viral encephalitis. The correlation between the two variables was analyzed. Results The expressions of P - selection on the surface of platelet membrane on less than 5 days and on 2 weeks after the onset of viral encephalitis were significantly higher in critical patients than those in normal control children and light patients( P

Objective To investigate the clinical features,recanalization after medical treatment and clinical outcome of cerebral artery dissection.Methods We reviewed the clinical records of ischemic stroke patients with cerebral artery dissection who visited the First Affiliated Hospital of Zhejiang University between October 2010 and December 2013.We recorded patients' general information,neurological deficit,imaging and clinical treatment.We followed up the patients and statistically analyzed demographic data,recanalization and clinical outcome.Results Among 28 cases of cerebral artery dissection,carotid dissection was the most common (n =19,67.9％),followed by vertebral artery dissection (n =7,25.0％),while the combination of carotid and vertebral artery dissection was rare (n =2,7.1％).In the imaging of cerebral artery dissection,wire beads and occlusion were the most common signs (n =15,53.6％),followed by rat tail sign (n =7,25.0％),aneurysmal dilatation (n =4,14.3％) and dualchamber levy (n =2,7.1％).There were 18 cases of acute cerebral infarction in the 28 studied cases,but there was no significant difference between the degree of stenosis and cerebral infarction caused by cerebral artery dissection.All patients received standard antithrombotic therapy.The difference of the recanalization rate between using anti-platelet aggregation and anticoagulant therapy was not statistically significant (5/6 vs 9/13,P =0.37).Conclusions Although cerebral artery dissection is relatively rare clinically,the typical clinical manifestations of the disease and the characteristic imaging are helpful for the diagnosis.Standardization of antithrombotic treatment is still the first-line treatment of cerebral artery dissection.

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Objective:To explore the efficiency of a new scoring system of gastric cancer screening for early gastric cancer in health examination population.Methods:The risk score of gastric cancer was assessed based on the new scoring system in health examination population. A notice for further gastroscopy was sent to the medium-risk and high-risk people. Gastroscopy was performed on those who agreed to undergo the examination.Results:From January to April 2019, a total of 5 357 people in health system visited the Physical Examination Center of Shanghai Songjiang Clinical Medical College of Nanjing Medical University for health examination. Seven hundred and forty people were classified as medium- and high-risk groups by the new screening system, 576 in medium-risk group, and 164 in high-risk group. Among them, 131 cases (17.70%) came for further gastroscopy, of whom 91 (69.47%) were in the medium-risk group and 40 (30.53%) in the high-risk group. After gastroscopy, 4 cases of gastric cancer and 1 case of esophageal cancer were detected, and both were early cancer. In the medium-risk group, 2 cases (2/91, 2.20%) of early gastric cancer and 1 case (1/91, 1.10%) of early esophageal cancer were found. In the high-risk group, 2 cases (2/40, 5.00%)of early gastric cancer were found. The tumor detection rate of high-risk group (5.00%) was higher than that of medium-risk group (3.30%), but there was no significant difference ( P>0.05). Conclusion:Risk stratification with the new scoring system of gastric cancer screening can improve the detection rate of early gastric cancer.