Risk factors of linezolid-related thrombocytopenia in patients in the department of hepatobiliary surgery / 药学实践杂志

Objective To provide the evidence for clinical medication safety by the investigation of the risk factors of linezolid-related thrombocytopenia in cancer patients in the department of hepatobiliary surgery. Methods Patients who received linezolid for anti-infective treatment from January 2017 to December 2021 were selected. The patients were divided into thrombocytopenia group and non-thrombocytopenia group according to whether thrombocytopenia occurred or not after administration of linezolid. The general data and laboratory indicators of the two groups were compared, and the risk factors of linezolid-related thrombocytopenia were screened by multivariate logistic regression analysis. Results A total of 104 patients were included in the study, including 84 patients who underwent surgery and 20 patients who did not. The incidence of linezolid-related thrombocytopenia was 24.0%. There were significant differences in gender, age, duration of linezolid use, platelet count, white blood cell count, alanine aminotransferase(ALT), aspartate aminotransferase(AST), total bilirubin, creatinine, estimated glomerular filtration rate between the two groups (P<0.05); logistic regression analysis suggested that age ≥60 years (OR=7.093; P=0.017), duration of linezolid use ≥12 days (OR=4.399; P=0.035), baseline platelet count ≤200×109/L (OR=8.470; P=0.004), baseline AST≥50 U/L (OR=15.465; P<0.001), and baseline white blood cell count ≥11×109/L (OR=11.436; P=0.001) were the risk factors for linezolid-related thrombocytopenia in cancer patients. Conclusion During the treatment of linezolid in cancer patients, attention should be paid to the adverse reactions of thrombocytopenia in the patients, especially those with old age, long-term treatment, low baseline platelets, poor baseline liver function, and high baseline white blood cell counts.

Molecular dynamics simulation study of effects of key mutations in SARS-CoV-2 on protein structures

SARS-CoV-2 has been spreading rapidly since 2019 and has produced large-scale mutations in the genomes. The mutation in genes may lead to changes in protein structure, which would have a great impact on the epidemiological characteristics. In this study, we selected the key mutations of SARS-CoV-2 from a real-time monitoring tool, including D614G, A222V, N501Y, T716I, S982A, D1118H of spike (S) protein, and performed molecular dynamics (MD) simulations on single-site mutant D614G, double-site mutant D614G&A222V and penta-site mutant N501Y&D614G&T716I&S982A&D1118H to investigate their effects on protein structure and stability using molecular dynamics (MD) simulations. The results suggested that D614G improved the stability of S protein, while D614G&A222V and N501Y&D614G&T716I&S982A&D1118H showed an increased solvent accessible surface area and they might enhance the ability of protein to react with the outside environment. Our findings could complement the mechanistic link between genotype--phenotype--epidemiological characteristics in the study of SARS-CoV-2. We also found no significant difference between the antigenicity of S protein and the mutants through Ellipro, which may reference for vaccine development and application.

Real-time monitoring epidemic trends and key mutations in SARS-CoV-2 evolution by an automated tool

With the global epidemic of SARS-CoV-2, it is important to monitor the variation, haplotype subgroup epidemic trends and key mutations of SARS-CoV-2 over time effectively, which is of great significance to the development of new vaccines, the update of therapeutic drugs, and the improvement of detection reagents. The AutoVEM tool developed in the present study could complete all mutations detections, haplotypes classification, haplotype subgroup epidemic trends and key mutations analysis for 131,576 SARS-CoV-2 genome sequences in 18 hours on a 1 core CPU and 2G internal storage computer. Through haplotype subgroup epidemic trends analysis of 131,576 genome sequences, the great significance of the previous 4 specific sites (C241T, C3037T, C14408T and A23403G) was further revealed, and 6 new mutation sites of highly linked (T445C, C6286T, C22227T, G25563T, C26801G and G29645T) were discovered for the first time that might be related to the infectivity, pathogenicity or host adaptability of SARS-CoV-2. In brief, we proposed an integrative method and developed an efficient automated tool to monitor haplotype subgroup epidemic trends and screen out the key mutations in the evolution of SARS-CoV-2 over time for the first time, and all data could be updated quickly to track the prevalence of previous key mutations and new key mutations because of high efficiency of the tool. In addition, the idea of combinatorial analysis in the present study can also provide a reference for the mutation monitoring of other viruses.

Evolution and molecular characteristics of SARS-CoV-2 genome

ObjectivesTo reveal epidemic trend and possible origins of SARS-CoV-2 by exploring its evolution and molecular characteristics based on a large number of genomes since it has infected millions of people and spread quickly all over the world. MethodsVarious evolution analysis methods were employed. ResultsThe estimated Ka/Ks ratio of SARS-CoV-2 is 1.008 or 1.094 based on 622 or 3624 SARS-CoV-2 genomes, and the time to the most recent common ancestor (tMRCA) was inferred in late September 2019. Further 9 key specific sites of highly linkage and four major haplotypes H1, H2, H3 and H4 were found. The Ka/Ks, detected population size and development trends of each major haplotype showed H3 and H4 subgroups were going through a purify evolution and almost disappeared after detection, indicating H3 and H4 might have existed for a long time, while H1 and H2 subgroups were going through a near neutral or neutral evolution and globally increased with time. Notably the frequency of H1 was generally high in Europe and correlated to death rate (r>0.37). ConclusionsIn this study, the evolution and molecular characteristics of more than 16000 genomic sequences provided a new perspective for revealing epidemiology of SARS-CoV-2.

Prediction and Evolution of B Cell Epitopes of Surface Protein in SARS-CoV-2

The discovery of epitopes is helpful to the development of SARS-CoV-2 vaccine. The sequences of the surface protein of SARS-CoV-2 and its proximal sequences were obtained by BLAST, the sequences of the whole genome of SARS-CoV-2 were obtained from the GenBank. Based on the NCBI Reference Sequence: NC_045512.2, the conformational and linear B cell epitopes of the surface protein were predicted separately by various prediction methods. Furthermore, the conservation of the epitopes, the adaptability and other evolutionary characteristics were also analyzed. 7 epitopes were predicted, including 5 linear epitopes and 2 conformational epitopes, one of the linear and one of the conformational were coincide. The epitope D mutated easily, but the other epitopes were very conservative and the epitope C was the most conservative. It is worth mentioning that all of the 6 dominated epitopes were absolutely conservative in nearly 1000 SARS-CoV-2 genomes, and they deserved further study. The findings would facilitate the vaccine development, had the potential to be directly applied on the treatment in this disease, but also have the potential to prevent the possible threats caused by other types of coronavirus.

Identification and Validation of Circulating MicroRNA Signatures for Breast Cancer Early Detection Based on Large Scale Tissue-Derived Data / 한국유방암학회지

PURPOSE: Breast cancer is the most commonly occurring cancer among women worldwide, and therefore, improved approaches for its early detection are urgently needed. As microRNAs (miRNAs) are increasingly recognized as critical regulators in tumorigenesis and possess excellent stability in plasma, this study focused on using miRNAs to develop a method for identifying noninvasive biomarkers. METHODS: To discover critical candidates, differential expression analysis was performed on tissue-originated miRNA profiles of 409 early breast cancer patients and 87 healthy controls from The Cancer Genome Atlas database. We selected candidates from the differentially expressed miRNAs and then evaluated every possible molecular signature formed by the candidates. The best signature was validated in independent serum samples from 113 early breast cancer patients and 47 healthy controls using reverse transcription quantitative real-time polymerase chain reaction. RESULTS: The miRNA candidates in our method were revealed to be associated with breast cancer according to previous studies and showed potential as useful biomarkers. When validated in independent serum samples, the area under curve of the final miRNA signature (miR-21-3p, miR-21-5p, and miR-99a-5p) was 0.895. Diagnostic sensitivity and specificity were 97.9% and 73.5%, respectively. CONCLUSION: The present study established a novel and effective method to identify biomarkers for early breast cancer. And the method, is also suitable for other cancer types. Furthermore, a combination of three miRNAs was identified as a prospective biomarker for breast cancer early detection.

Monitoring of antimicrobial resistance of Acinetobacter baumannii in the intensive care unit of a hospital from 2010 to 2013 / 中国感染控制杂志

Objective To analyze the isolation rates and antimicrobial resistance of Acinetobacter baumannii (AB) from intensive care unit (ICU)between 2010 and 2013,and provide evidence for clinical anti-infective therapy. Methods The isolation and antimicrobial resistance of AB from ICU between 2010 and 2013 were analyzed retro-spectively.Results A total of 1 413 pathogenic strains were isolated,556(39.35%)of which were AB,isolation rates in each year were 39.45%,41 .35%,29.44%,and 40.53% respectively.AB were mainly isolated from lower respiratory tract (75.72%).Antimicrobial susceptibility testing results showed that AB had low resistance rates to cefoperazone/sulbactam(5.85%)and amikacin (17.45%);detection rates of multidrug-resistant and extensively drug-resistant AB increased from 9.63% and 3.70% to 42.50% and 31 .88%,respectively (both P < 0.001 ). Conclusion AB is the common pathogen in ICU,antimicrobial resistance is serious,isolation of multidrug-resistant and extensively drug-resistant AB increased year by year;intensifying the monitoring of drug resistance is helpful for the treat-ment and prevention of AB infection.

Mechanism by which interferon reduces the resistance of MGMT positive glioma stem cells to temozolomide / 中国组织工程研究

OBJECTIVE:To investigate the role of interferon to increase the sensibilization of MGMT positive glioma stem cel s to temozolomide in vitro. METHODS:Glioma cel lines, U251 and SKMG-4, were induced by suspended cloning bal formation method to harvest MGMT positive glioma stem cel s, U251G and SKMG-4G. Cel counting kit-8 assay was used to detect the kil ing effect of interferonα/βcombined with temozolomide on MGMT positive glioma stem cel s. RT-PCR and western blot assay were employed to determine the expression of MGMT and nuclear factorκB in MGMT positive glioma stem cel s. RESULTS AND CONCLUSION:Western blot results showed positive expression of MGMT in U251G and SKMG-4G cel s at protein levels. After intervention with interferonα/β, the mRNA expression of MGMT and nuclear factorκB in SKMG-4G and U251G cel s was reduced significantly, and then further decreased after temozolomide treatment. These findings indicate that interferonα/βcan remarkably strengthen the kil ing effect of temozolomide on MGMT positive glioma stem cel s.

Research on the technological process and methods of the comprehensive pharma-ceutical experiment of rutin tablets / 药学实践杂志

Objective To imitate the development process of new drugs with rutin as the model and to do multidiscipli-nary experiments of preparation and pharmacodynamics of rutin tablets .Methods Thin-layer chromatography was used to i-dentify rutin in Pagodatree flower bud .High-performance liquid chromatography was used for quantitative determination of ru-tin in Pagodatree flower bud and rutin products .The vasodilatation effect of rutin was investigated .The preparation of rutin tablets was completed .Results and Conclusion We completed the identification of Pagodatree flower bud ,extraction and puri-fication of rutin from Pagodatree flower bud ,the assay of rutin ,the pharmaco-dynamics study and the formulation of rutin tab-lets .The experiments helped the postgraduates to be familiar with the research process of new drugs and to improve their ex-perimental operation skills .

Screening of genetic markers based on human type 2 diabetes mellitus (T2DM) susceptibility loci in a Cynomolgus T2DM model / 中国比较医学杂志

Objective Type 2 diabetes mellitus ( T2DM) as a common disease around the world becomes a great threat to the health of human beings.The cynomolgus T2DM model, which preferably simulates human T2DM onset and progress, can be beneficial to the drug development and clinical treatment.In the present study, 37 of T2DM-susceptibility SNPs and the extended genome sequences were used to obtain corresponding SNPs in the T2DM cynomolgus monkeys. Methods Firstly, DNA pool screening was conducted.Then, using polymerase chain reaction to amplify and to sequence the cynomolgus homologous sequences.Using DNAStar software to analyze the differences between bases.Finally, we used analysis of variance and F test to calculate the frequency of alleles.We also used the GLM models of SAS software to analyze the association of genotype with fasting plasma glucose and glycosylated hemoglobin.Results SNP661A,SNP661B, SNP343A, SNP343B, SNP343C, SNP565A, SNP565B and SNP565C were found to have a significant difference of allele frequencies between spontaneous cases and controls.Conclusions The findings of this study suggest that SNP661A, SNP661B, SNP343A, SNP343B and SNP343C may play an important role in the establishment of cynomolgus T2DM models.

Mutational research on the role of lysine 21 in the Pichia stipitis xylose reductase / 生物工程学报

The xylose reductase of Pichia stipitis is one of the most important enzymes. It can be used to build up recombinant Saccharomyces cerevisiae strain for utilizing xylose and producing ethanol. Intercellular redox imbalance caused by NADPH preference over NADH for Pichia stipitis xylose reductase (PsXR) has been considered to be one of the main factors for poor ethanol productivity. Some key amino acids of PsXR, which affect the activity or coenzyme preference, were investigated in our previous study. In this study, Lys21 were rational designed for site-directed mutagenesis to alter coenzyme specificity of PsXR from NADPH and NADH into single NADH. The wild gene and mutagenesis genes were ligated into pET28b, and were transferred into E.coli BL21(DE3). After induced by IPTG, the xylose reductases were purified. Purified mutants K21A (Lys21-->Ala), K21R(Lys21-->Arg) were characterized by steady-state kinetic analysis. The results showed that the coenzyme dependence of K21A was completely reversed to NADH.