Identification of lysine acetyltransferase p300 substrates using 4-pentynoyl-coenzyme A and bioorthogonal proteomics.

Proteomic studies have identified a plethora of lysine acetylated proteins in eukaryotes and bacteria. Determining the individual lysine acetyltransferases responsible for each protein acetylation mark is crucial for elucidating the underlying regulatory mechanisms, but has been challenging due to limited biochemical methods. Here, we describe the application of a bioorthogonal chemical proteomics method to profile and identify substrates of individual lysine acetyltransferases. Addition of 4-pentynoyl-coenzyme A, an alkynyl chemical reporter for protein acetylation, to cell extracts, together with purified lysine acetyltransferase p300, enabled the fluorescent profiling and identification of protein substrates via Cu(I)-catalyzed alkyne-azide cycloaddition. We identified several known protein substrates of the acetyltransferase p300 as well as the lysine residues that were modified. Interestingly, several new candidate p300 substrates and their sites of acetylation were also discovered using this approach. Our results demonstrate that bioorthogonal chemical proteomics allows the rapid substrate identification of individual protein acetyltransferases in vitro.

Quality evaluation of Cyclocarya paliurus from different habitats by fingerprint and multi-component quantification combined with chemical pattern recognition / 中草药

Objective: Based on the method of UPLC fingerprint, multi-component quantification and chemical pattern recognition, the quality of Cyclocarya paliurus from different producing areas was evaluated to provide basis for further development and utilization. Methods: The UPLC fingerprints of 20 batches of C. paliurus from different habitats in five provinces were established to determine the common peaks. Three chemical components were identified and the content of the samples was determined by comparison with the reference materials. Cluster analysis and principal component analysis were carried out by SPSS 22.2 and SIMCA software. Results: A total of 16 chromatographic peaks were selected as the common peaks of the fingerprint, 20 batches of C. paliurus could be divided into six categories by cluster analysis, and the results of principal component analysis and cluster analysis were basically the same; By principal component analysis, the cumulative variance contribution rate of the five principal component factors was 86.765%. The quality of S14 and S15 samples from Enshi, Hubei Province was the best, and S4, S5, S8, and S16 of Xiushui, Jiangxi Province were the second. The content of isoquercetin, quercitrin and afzelin were 0.360%-0.884%, 0.263%-1.097%, and 0.092%-0.403%, respectively, among which the total content of S4, S8, S14, and S15 were the top 4, and that of S18 and S20 was in the last two places. Conclusion: Combined with fingerprint, cluster analysis and principal component analysis, we can evaluate the quality of C. paliurus more comprehensively. Isoquercetin, quercitrin and afzelin can be used as the index components for C. paliurus quality control.

Polio antibody levels in infants during 2015-2016 in Shanghai before and after polio vaccination strategy adjustment / 上海预防医学

Objective To investigate the neutralized antibody level of poliomyelitis among healthy children before and after implementation of the new immunization strategy in Shanghai. Methods The poliomyelitis levels of healthy children were detected by neutralization test of the micro cells. Results The neutralizing antibody positive rates were 100% for polioⅠand polioⅢ before and after implementation of the new immunization strategy.GMTs (geometric mean titers) for polioⅠwere 1:1 047.46 before it, and 1:254.70 after it; those for polio Ⅲ were 1:513.52 before it and 1:185.49 after it. Conclusion The neutralized antibody levels of the poliomyelitis among healthy infants maintain a high level before and after implementing the new immunization strategy in Shanghai.

Spectrum-effect relationship between UPLC fingerprint of Smilax china and anti-pelvic inflammation in rats / 中国中药杂志

To study the correlation between ultra high performance liquid chromatography( UPLC) fingerprint of Smilax china and its anti-pelvic inflammatory effect,and to explore the pharmacodynamic material basis of S. china against pelvic inflammatory disease.UPLC fingerprints of 10 batches of S. china from different habitats were established,and the values of SOD,MDA,TNF-α,and IL-6 in rats with pelvic inflammation were measured. The weight of each single pharmacodynamics index to the total efficacy was determined by analytic hierarchy process,and the contribution of each peak in fingerprints to the each single pharmacodynamics index and total efficacy was analyzed by the grey relational analysis. Then the structures of chemical constituents at the identified peaks were confirmed by comparing with the reference substance. The 27 common characteristic peaks of UPLC fingerprints were all related to the anti-pelvic inflammation effect of S. china,of which 13 peaks were identified as peak 2( 3,5-dihydroxy-2-methylbenzoic acid-3-O-glucoside),peak 3( chlorogenic acid),peak 5( 2,7,4-trihydroxydihydroflavone-5-O-glucoside),peak 6( 7,4-dihydroxydihydroflavonol-5-O-glucoside),peak 7( taxifolin-7-O-glucoside),peak 9( taxifolin),peak 10( polydatin),peak 11( oxyresveratrol),peak 12( astilbin),peak15( resveratrol),peak 16( quercitrin),peak 18( engeletin) and peak 24( kaempferol). The correlation degree of 21 peaks and the total efficacy was greater than 0. 8,and the top 10 ranked by correlation degree were as follows: peak 1,3,7,19,18,17,4,11,16,and 21. The results showed that the anti-pelvic inflammation effect of S. china was achieved by the combined action of pharmacodynamic substances. In order to control the quality of S. china and its prepared slices more effectively,the index components of content detection should be selected reasonably.

Efficacy and safety of integrative medical program based on blood cooling and detoxification recipe in treating patients with hepatitis B virus related acute-on-chronic liver failure: a randomized controlled clinical study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy and safety of integrative medical program based on blood cooling and detoxification recipe (BCDR) in treating patients with hepatitis B virus related acute-on-chronic liver failure (HBV-ACLF) of heat-toxicity accumulation syndrome (HTAS).</p><p><b>METHODS</b>Adopting randomized controlled clinical design, a total of 105 HBV-ACLF patients of HTAS were randomly assigned to the trial group (64 cases) and the control group (41 cases). Patients in the control group were treated with comprehensive Western therapy, while those in the trial group were treated with comprehensive Western therapy plus BCDR. All were treated for 8 weeks and followed up for 40 weeks. Effect and safety of the treatment were assessed, including fatality, liver functions [total bilirubin (TBIL), albumin (ALB), alanine aminotransferase (ALT), and aspartate transaminase (AST)], and prothrombin activity (PTA) after treatment and at week 48 of follow-ups.</p><p><b>RESULTS</b>After 8-week treatment, there was statistical difference in the overall fatality rate (15.63% vs 34.15%), the fatality rate in the mid-term (25.0% vs 64.7%), TBIL at week 8 (64.54 +/- 79.75), AST [at week 2: (178.97 +/- 44.24) U/L vs (288.48 +/- 58.49) U/L; at week 4: (61.65 +/- 27.36) U/L vs (171.12 +/- 89.11) U/L] and PTA [at week 4: (58.30 +/- 15.29) vs (42.56 +/- 15.27); at week 6: (60.77 +/- 20.40) vs (43.08 +/- 12.79)] (all P < 0.05). At week 48 of the followup, the fatality rate of the trial group (21.88%) decreased by 17. 14% when compared with that of the control group (39.02%; P < 0.05). No obvious adverse event occurred in the two groups during the 8-week treatment period.</p><p><b>CONCLUSION</b>BCDR could significantly reduce the mortality of HBV-ACLF patients.</p>

Molecular epidemiological analysis of rubella virus isolates from 2001 to 2011 in Shanghai, China / 病毒学报

Throat swabs collected from patients whose serum was measles IgM negative and rubella IgM positive during 2001-2011 were used to conduct cell culture for rubella virus. After identification of cell culture with RT-PCR, nucleotide of gene E1 of rubella virus was amplified and sequenced, followed by molecular epidemiological analysis. A total of 31 rubella viruses were isolated from 60 throat swabs. Compared 27 isolates with the WHO reference strains of all genotypes, phylogenetic tree was constructed based on the amplified 739 nucleotide fragment. These isolates belonged to two different genotypes respectively. Isolates 11009, 11052 and 11106 in 2011 belonged to genotype 2B, and others belonged to genotype 1E. Most of mutations were nonsense mutation, and sequence of amino acid was highly conserved. Amino acid sequence of most isolates of genotype 1E was identical, which suggested rubella viruses from same transmission chain might be transmitted continually since 2001. Rubella virus genotype 2B was found to be popular for the first time in Shanghai in 2011. The nucleotide sequences of these genotype 2B isolates showed 99% identity compared with that of isolates recently from Vietnam, Japan and Argentina. The resources of these strains were not confirmed due to the absence of rubella virus surveillance before.

Expression of intrahepatic CD4+ CD25+ regulatory T cells in patients with chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the expression and distribution of intrahepatic CD4+ CD25+ regulatory T cells in immuno-tolerant and immuno-clearance phase of patients with chronic hepatitis B.</p><p><b>METHODS</b>The expression of FoxP3 was detected in 19 cases of immuno-tolerant phase and 12 cases of immuno-clearance phase by immunohistochemistry. The relation between the intrahepatic expression of FoxP3 and the clinicopathological features were analyzed.</p><p><b>RESULTS</b>The positive signal of FoxP3 is located in nuclear of lymphocyte and mainly aggregated in portal areas as well as occasionally scattered in hepatic sinusoids. The expression of intrahepatic FoxP3 in the group of immuno-tolerant phase was significantly increased than those in normal control (P < 0.01), and greatly decreased than those in immuno-clearance phase (P < 0.01). No correlation was observed among the expression of intrahepatic FoxP3, ALT, levels of HBV DNA, HBeAg positive, in patients of immuno-clearance phase, respectively. There were significant differences between immuno-tolerant phase and immuno-clearance phase age, ALT, TBIL, PTA, HBV-DNA and detection of HBeAg but not in sex and family history of HBV infection.</p><p><b>CONCLUSION</b>CD4+ CD25+ regulatory T cells may play important roles in the clearance of HBV as well as in liver inflammation and injury during chronic HBV infection.</p>

Clinical experience from treatment of seven SARS patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To summarize the clinical experience from treatment of patients with severe acute respiratory syndrome (SARS).</p><p><b>METHODS</b>Retrospective analysis of seven patients with SARS in Ditan hospital treated since April 22 in 2004 was performed.</p><p><b>RESULTS</b>In the 7 patients, 2 were male, 5 were female, and the average age was (35.3 plus/minus 11.3) years. The main clinical manifestations were fever, cough, minor or serious dyspnea, nausea, signs of injury to other organs, and so on. The treatment regiments included oxygen, small dosage and short period of methylprednisolone (1 to 2 mg/kg), use of ventilator, psychological intervention, and treatment of underlying diseases, after which, all the 7 patients recovered.</p><p><b>CONCLUSION</b>Rational use of methylprednisolone and timely use of ventilator were the key steps of treatment.</p>