Proteolytic cleavage is one of the post-translational modifications and plays important roles in many biological processes, such as apoptosis and tumor cell metastasis. The identification of the cleavage events can improve our understanding of their biological functions in these processes. Although proteomic approaches using N-terminal labeling have resulted in the discovery of many proteolytic cleavages, this strategy has its own inherent drawbacks. Labeling of protein C-termini is an alternative approach. Here, we optimized the labeling procedure in the profiling protein C-termini by enzymatic labeling (ProC-TEL) and improved the labeling efficiency for the positive isolation of protein C-terminal peptides and mass spectrometric identification. We applied this approach to a complex protein mixture from Escherichia coli and identified many C-terminal peptides and internal cleaved peptides from more than 120 proteins. From the identified cleavages, we found several previously known internal proteolytic cleavage sites and many novel ones which may play roles in regulating normal biological processes. This work provides a potential new way, complementary to the N-terminomics, for the identification of proteolytic cleavages in complex biological systems.
Cathepsin A ; chemistry ; Protein C ; chemistry ; Protein Processing, Post-Translational ; Proteolysis ; Proteomics

Objective To investigate how the independent components(ICs)energies of multichannel local field potentials(LFPs) code event base on the analysis of ICA of the cortical LFPs of rats. Methods Taking the event point as the zero point, 15-channel LFPs between the span of ±500ms recorded from the prefrontal cortex of rats were decomposed into 15 ICs. The energies of the ICs were computed in a 50-ms window. By sliding the window with step of 25 ms, a dynamic distribution mapping of the 15 ICs' energies was established. ICs with distinctly increased energies during the span of ±200 ms, which indicating that these ICs energies coded event,were selected as the targets. The corresponding channels of these ICs were determined consequently via the inverse transformation of ICA. Results Considering each trail of the repetitious analysis for the same segment of data, the spatial localization of the dominate function region(s) turned out to be relatively stable in spite of the uncertainty of the number and sequence of the target IC(s) due to the ambiguities of the decomposition of ICA.Meanwhile, the analysis results of a series of data segments showed satisfactory correspondence between data segments and dominate function regions. Conclusion The ICs' energies of multichannel LFPs are able to code events in working memories; It is valid for ICA to identify the coding patterns of multichannel LFPs to events; ICA is capable to localize the dominate function regions of event coding with satisfactory robustness.

Objectives To explore the intrinsic factors related to the pathogenesis of acute arterial thrombosis (AAT) and to elucidate the patho-genesis of AAT on the basis of differentially expressed genes. Methods Patients with acute myocardial infarction (AMI), stable angina (SA) and healthy controls (n=20 per group) were recruited, and the whole human genome microarray analysis was performed to detect the dif-ferentially expressed genes among these subjects. Results Patients with AMI had disease-specific gene expression pattern. Biological func-tional analysis showed the function of T cells was significantly reduced, the mitochondrial metabolism significantly decreased, the ion me-tabolism was abnormal, the cell apoptosis and inflammatory reaction increased, the phagocytosis elevated, the neutrophil-mediated immunity increased and the post-traumatic repair of cells and tissues increased in AMI patients. The biological function in SA group and healthy con-trols remained stable and was comparable. Conclusions The reduced function of T cell gene models in AAT showed the dysfunction of the immune system. The pathogenesis of AAT may be related to the inflammatory reaction after arterial intima infection caused by potential pathogenic microorganisms.

Objective To study the clinical efficacies of different surgical palliative treatments for patients with advanced pancreatic carcinoma.Methods A retrospective analysis was conducted on the clinical data of 119 patients with advanced pancreatic carcinoma who were treated in the department of General Surgery of Linyi's Central Hospital from January 2010 to January 2016.According to the different surgical methods the patients received,they were divided into the seed group (n =38),the radiofrequency ablation (RF) group (n =52) and the control group (n =29).Patients' general data,operation data,postoperative complications and follow-up profiles were analyzed.Results There was no significant difference in the patients'general data among the three groups of patients (P ＞ 0.05).The incidences of postoperative complication in the seed group,the RF group and the control group were 55.3％,38.5％ and 20.7％,respectively.There was significant differences between the incidences of postoperative complications between the seed group and the control group (P ＜ 0.05).There was no perioperative death.The postoperative follow-up rate was 91.6％.Pain in the seed group and the RF group significantly improved,while it was not significantly improved in the control group.There were significant differences in the postoperative pain scores (P ＜0.05).The 6 months,1-year and 2-year overall survival rates were 61.3％,25.9％,8.6％ respectively.The median survival was 8 months.In the seed group,the 6 months,1-year and 2-year survival rates were 75.2％,37.8％,18.9％ respectively.The median survival was 9 months.In the RF group,the 6 months,1-year and 2-year survival rates were 60.4％,25％,6.3％ respectively.The median survival was 8 months.In the control group,the 6 months,1-year and 2-year survival rates were 53.4％,15.3％,3.8％ respectively.The median survival was 6.5 months.On log-rank test,there was no significant difference in survival rates among the three groups (P =0.145).Conclusions Patients with advanced pancreatic carcinoma had poor prognosis.I125 radioactive particles implantation and radiofrequency ablation were useful to improve patients'quality of life and prolonged their survival.

Objective:To analyse the role of Bcl-6/Blimp-1/IL-21 in pathogenesis of primary Sj?gren′s syndrome(PSS),the relationship between Bcl-6/Blimp-1/IL-21 expression and labial gland biopsy grading.Methods:Immunohistochemical(IHC) method was used to detect the expression of Bcl-6/Blimp-1 in salivary gland between 30 cases pations with PSS in disease group and 11 cases patients with mucous cyst,lower lip trauma in control group,and the serum IL-21 levels between disease group of 40 cases patients with PSS and 30 cases healthy volunteers were measured by enzyme-linked immunosorbent assay( ELISA) ,relations among 15 patients with IL-21 expression levels with labial gland biopsy grading.Results:The expression levels of Bcl-6/Blimp-1/IL-21 in disease group were higher than control group;in disease group,with pathological grades increased,the expression levels of Bcl-6/Blimp-1/IL-21 were also raised.Conclusion:①Bcl-6/Blimp-1/IL-21 may participate in the pathogenesis of PSS;Bcl-6/Blimp-1/IL-21 is associated with infiltration lymphocytes of salivary gland.

Objective To investigate expression differences of neutrophil and mononuclear phagocyte related gene mRNAs among acute myocar-dial infarction (AMI), stable angina (SA) and control groups, and then discuss their expression characteristics in the stable angina pectoris (SAP) and AMI stages of coronary artery disease (CAD). Methods Whole Human Genome Oligo Microarrays were applied to assess the differential expression characteristics of neutrophil and mononuclear phagocyte related mRNAs in patients with AMI (n=20), SA (n=20) and controls (n=20). Results (1) Almost all colony-stimulating factors (CSF) and their receptors related mRNAs was up-regulated in AMI and SA groups compared with the control group, and the expression of granulocyte-macrophage colony stimulating factor receptor (GM-CSFR) and granulocyte colony stimulating factor receptor (G-CSFR) mRNAs in the AMI group was significantly up-regulated com-pared with the other two groups (P<0.01). (2) The expression of mRNAs related to monocyte chemoattractant protein-1 (MCP-1), CCR2 (MCP-1 receptor) and CXCR2 (IL-8 receptor) was significantly up-regulated (P<0.01) in AMI group compared with SA and control groups. IL-8 mRNA expression in the AMI group was clearly higher than the controls (P<0.05). (3) All mRNAs expression related to opsonic re-ceptors (IgG FcR and C3bR/C4bR) was significantly up-regulated in AMI group compared with SA and control group (P<0.01), and the SA group showed an upward trend compared with controls. (4) Most pattern recognition receptor (PRR)-related mRNAs expression was up-regulated in AMI group compared with SA and control groups. Most toll-like receptor (TLR) mRNAs expression was significantly up-regulated (P<0.01) than the SA and control groups;macrophage scavenger receptor (MSR) mRNA was significantly up-regulated in AMI group compared with the control group (P<0.01), and the SA group showed an upward trend compared with the controls. Conclusions The expression of most neutrophil and mononuclear-macrophage function related genes mRNAs was significantly up-regulated by stages during the progression of CAD, suggesting that the adhesive, chemotactic and phagocytic functions of neutrophil and mononuclear-ma-crophage were strengthened in the occurrence and development of coronary atherosclerosis and AMI. This also showed a stepped up-ward trend as the disease progressed.

Objective:The study aimed to study the efficacy and safety of combined dual therapy using anti-programmed death (PD)-1 and tyrosine kinase inhibitor (TKI) with combined triple therapy using anti-PD-1, TKI and locoregional intervention triple therapy in patients with postoperative refractory recurrent liver cancer.Methods:Patients with postoperative refractory recurrent liver cancer who had undergone either anti-PD-1 and TKI dual therapy or anti-PD-1, TKI and locoregional intervention triple therapy between July 2016 and March 2019 at the First Medical Center, Chinese PLA General Hospital were retrospectively studied. Tumor responses were assessed by the modified response evaluation criteria in solid tumors and overall survival and progression free survival were compared. Adverse events were evaluated according to the National Cancer Institute Common Terminology Criteria for Adverse Events.Results:Of 63 patients who were included in this study, there were 25 patients in the dual therapy group (16 males and 9 females, aged 54.3±8.8 years) and 38 patients in the triple therapy group (31 males and 7 females, aged 55.5±8.4 years). The 1-year survival rate of the triple therapy group was significantly higher than the dual therapy group (94.5%vs 54.9%) ( P<0.01). The disease control rate was 64.0% (16/25) in the dual therapy group and 84.2% (32/38) in the triple therapy group, and the difference was not significant ( P>0.05). The incidence of treatment-related adverse events in the triple therapy group and the dual therapy group were 78.9% (30/38) and 80% (20/25), respectively. There was no treatment-related death in the 2 groups. Conclusions:Anti-PD-1 and TKI dual therapy and anti-PD-1, TKI and locoregional intervention triple therapy were effective and tolerable treatments for postoperative refractory recurrent liver cancer. The latter treatment had a significantly better clinical benefit on survival outcomes.

Objective To summarize the diagnosis and treatment of congenital pulmonary airway malformation in children.Methods The clinical data of 68 children with congenital pulmonary airway malformations from June 2014 to June 2017 were analyzed retrospectively,including 39 males and 29 females with mean age(1.1 ±:0.5) years old(from 2 months 12 days to 15 years),7 cases of pulmonary cyst resection in 68 cases,3 cases with lung segment resection,1 cases with lingulectomy,57 cases with lobectomy.Results 68 cases were cured and discharged without death.Postoperative short-term complications included subcutaneous emphysema in 5 cases,pleural effusion in 2 cases,pneumothorax in 2 cases.All cases were followed up well between 3 months and 1 year.Conclusion The diagnosis of congenital pulmonary airway malformation in children mainly depends on imaging examination.Because it is easy to repeat infection,and can not rule out malignant transformation,early diagnosis and early treatment should be done.Surgical resection is the only way to treat the disease.

Objective:To compare the preventive effects of different doses of esketamine on remifentanil-induced hyperalgesia(RIH)in the patients underwent thoracoscopic pnlmonary lobectomy,and to provide the basis for the multimodal analgesia and rapid postoperative recovery.Methods:The prospective,double-blind,parallel-designed randomized controlled trial(RCT)were conducted,and 107 patients who underwent visual-assisted thoracoscopic pulmonary lobectomy were included.The patients were randomly divided into normal control group,low dose of esketamine group,and high dose of esketamine group using random number methods.Except the patients who were eliminated and those who dropped out of the study,there were 31 patients in normal control group,33 patients in low dose of esketamine group,and 33 patients in high dose of esketamine group.The patients in low dose of esketamine group were given the intravenous injection of 0.25 mg·kg-1 esketamine(diluted to 5 mL)during anesthesia induction;the patients in high dose of esketamine group were given the intravenous injection of 0.50 mg·kg-1 esketamine(diluted to 5 mL),and the patients in normal control group were given 5 mL intravenous injection of saline.The mechanical pain thresholds of the non-dominant forearm skin and skin around the surgical incision at different time points,numeric rating scale(NRS)scores,Ramsay sedation scores,perioperative analgesic drug dosages,and the incidences of adverse reactions such as postoperative delirium,nausea,and vomiting of the patients in various groups were recorded.Results:Compared with normal control group,the mechanical pain thresholds around the surgical incision skin of the patients in low and high doses of esketamine groups were increased(P＜0.05);compared with low dose of esketamine group,the extubation time of the patients in high dose of esketamine group was increased(P＜0.05).Two minutes after anesthesia induction administration,compared with normal control group,the mean arterial pressure(MAP)and heart rate(HR)of the patients in low and high doses of esketamine groups were increased(P＜0.05),but there were no significant differences in the MAP and HR of the patients between low dose of esketamine and high dose of esketamine group(P＞0.05);compared with normal control group,the incidences of hallucinations and delirium among the patients in high dose of esketamine group were increased(P＜0.05),while there were no significant differences in the incidences of above adverse reactions in low dose of esketamine group(P＞0.05);compared with low dose of esketamine group,the incidences of hallucinations and delirium among the patients in high dose of esketamine group were increased(P＜0.05).Conclusion:Intravenous administration of esketamine with a dosage of 0.25 mg·kg-1 during anesthesia induction improves the postoperative mechanical pain threshold of the patients undergoing thoracoscopic pulmonary lobectomy,which exhibits effective prevention of RIH without an increase in incidences of adverse reactions during the perioperative period.

Objective To explore the clinical efficacy of Chaiqi Ningshen Anmian Decoction(CNAD)combined with wrist ankle acupuncture(WAA)in treating chronic insomnia(CI)patients with heart and spleen deficiency.Methods CI patients diagnosed and treated at the Beijing Huairou Hospital of Traditional Chinese Medicine from April 2022 to April 2023 were selected,and patients were randomly divided into the Eszolam(ET)group and the combination group(CNAD combined with WAA)according to the random number table method,with 50 cases in each group.The primary outcome was clinical efficacy[evaluated by the Traditional Chinese Medicine Syndrome Integral(TCMSI)].Secondary outcomes included changes in cognitive function[assessed by the Montreal Cognitive Assessment(MoCA)],anxiety level[assessed by the Hamilton Anxiety Scale(HAMA)],sleep quality[assessed by the Pittsburgh Sleep Quality Index(PSQI)],glycated serum albumin(GA),serotonin(5-HT),interleukin-1 β(IL-1β),and C-reactive protein(CRP)before and after treatment in both groups.Results Before treatment,there was no significant differences in total TCMSI between the two groups(P＞0.05).After treatment,the total TCMSI of the combination group was significantly lower than that of the ET group(P＜0.05),and the treatment effectiveness rate was significantly higher in the combination group(P＜0.05).Before treatment,there were no significant differences in MoCA score,HAMA score,PSQI score,serum GA level,serum 5-HT level,serum IL-1 level and serum CRP level between the groups(P＞0.05).After treatment,the MoCA score,serum GA level,and serum 5-HT level in the combination group were significantly higher than those in the ET group(P＜0.05),while the HAMA score,PSQI score,serum IL-1β level,and serum CRP level were significantly lower in the combination group(P＜0.05).Conclusion Compared with ET,CNAD combined with WAA significantly improves insomnia symptoms in CI patients with heart and spleen deficiency,enhances cognitive function and sleep quality,and reduces anxiety levels.This may be related to the upregulation of serum 5-HT,IL-1 β and the inhibition of the inflammatory response.