Penicillin G acylases (PGAs) are industrially important enzymes primarily used for the synthesis of first- and second-generation cephalosporins or penicillins. This study aims to establish a high-efficiency biosynthetic system for cefradine on the purpose of significantly enhancing its catalytic efficiency in cefradine synthesis and developing its potentials for industrial application. In this study, we identified and engineered penicillin G acylase and obtained a highly active mutant KsPGA M7(M168F/F313G) for the synthesis of cefradine. The mutant achieved a conversion rate over 95% in the scaled-up reaction. To validate its industrial applicability, we immobilized both the wild-type and mutant enzymes and applied them in continuous flow reactions, which achieved a space-time yield of 2 800 g/(L·d). This study lays a foundation for the future applications of penicillin G acylases in the industrial synthesis of cefradine.
Penicillin Amidase/biosynthesis* ; Protein Engineering/methods* ; Cephradine/metabolism* ; Escherichia coli/metabolism* ; Enzymes, Immobilized/metabolism* ; Recombinant Proteins/biosynthesis*

Proteomic characterization of plasma is critical for the development of novel pharmacodynamic bio-markers.However,the vast dynamic range renders the profiling of proteomes extremely challenging.Here,we synthesized zeolite NaY and developed a simple and rapid method to achieve comprehensive and deep profiling of the plasma proteome using the plasma protein corona formed on zeolite NaY.Specifically,zeolite NaY and plasma were co-incubated to form plasma protein corona on zeolite NaY(NaY-PPC),followed by conventional protein identification using liquid chromatography-tandem mass spectrometry.NaY was able to significantly enhance the detection of low-abundance plasma proteins,minimizing the"masking"effect caused by high-abundance proteins.The relative abundance of middle-and low-abundance proteins increased substantially from 2.54％to 54.41％,and the top 20 high-abundance proteins decreased from 83.63％to 25.77％.Notably,our method can quantify approxi-mately 4000 plasma proteins with sensitivity up to pg/mL,compared to only about 600 proteins iden-tified from untreated plasma samples.A pilot study based on plasma samples from 30 lung adenocarcinoma patients and 15 healthy subjects demonstrated that our method could successfully distinguish between healthy and disease states.In summary,this work provides an advantageous tool for the exploration of plasma proteomics and its translational applications.

ObjectiveTo observe the effect of the enhanced external counterpulsation （EECP） combined with drug on social function and efficacy in patients with depressive episodes， so as to provide references for the treatment of depressive episodes. MethodsA total of 66 hospitalized patients who was in hospital at department of psychiatry of the Second Xiangya Hospital of Central South University， met the criteria of Diagnostic and Statistical Manual of Mental Disorders，fifth edition（DSM-5） diagnosis of depressive episode or bipolar disorder depressive episode from May 2019 to March 2020 were included by simple random sampling. The participants were divided into study group （n=36） and control group （n=30） according to the random number table method. Both groups received conventional drug treatment， and the study group recieved the EECP intervention at same time. The Depression symptoms and social function were assessed before and after treatment by using Hamilton Depression Scale-24 item （HAMD-24） and Sheehan Disability Scale （SDS）. Treatment efficacy of the two groups was compared. ResultsAfter the intervention， the HAMD-24 and SDS scores in both groups were lower than those before treatment， the differences were statistically significant （t=8.149， 5.791， 8.016， 3.488， P˂0.01）. And the SDS score of the study group was siginficantly lower than that of the control group （t=-3.008， P<0.05）. The total effective rate of treatment in the study group was higher than that of the control group， and the difference was statistically significant （90.63% vs. 63.33%， χ²=8.725， P˂0.05）. ConclusionEECP therapy combined with drug has better efficacy on the patients with depressive episodes， and it can improve social function effectively.

Objective:To establish a nomograph prediction model of early critical changes in children with febrile convulsion, and to provide guidance for the prevention and nursing of children with febrile convulsion.Methods:Convenient sampling method was adopted to select 384 children with febrile convulsion in Anhui Children ′s Hospital from January 2018 to April 2021 as the research objects. Based on pews, the children with febrile convulsion were divided into 334 cases of non risk group and 50 cases of risk group. Binary Logistic regression analysis were used to determine the independent risk factors affecting the early critical changes of children with febrile convulsion. A nomogram was drawn based on the independent risk factors. The discrimination and consistency of the model were verified by model ROC curve and Hosmer Lemeshow goodness of fit.The external validation of model prediction efficiency were verified by validation data. Results:Binary Logistic regression analysis showed that age, respiratory rhythm disorder, unconsciousness, breath rate, heart rate, neutrophil-to-lymphocyte ratio (NLR), red blood cell distribution width (RDW), duration of first convulsion and mean body temperature after first convulsion were influence factors for early critical changes in children with febrile convulsion ( P<0.05). The C-index of the model was 0.974 (95% CI 0.954-0.993), and the C-index of the external validation of the model was 0.922 (95% CI 0.880-0.966). The results of H-L fitting test showed that the difference was not statistically significant( χ2=0.29, P>0.05). Conclusions:The early critical changes of children with febrile convulsion may be affected by respiratory rhythm disorder, confusion of consciousness, breath rate, heart rate, NLR, RDW, duration of the first convulsion, mean temperature after the first convulsion and other factors. Pediatric emergency department should collect corresponding intervention measures for children with febrile convulsion according to the establishment of prediction model to prevent their early deterioration.

Objective:To investigate the applicability of time series model in predicting incidence of nosocomial infection in a cancer center in Shanghai, and to provide the references for early warning and prevention.Methods:The nosocomial infection data of inpatients of a tertiary oncology hospital in Shanghai from 2013 to 2018 were collected. The autoregressive integrated moving average (ARIMA) model and the exponential smoothing model were established by SPSS 22.0 expert modeler. The fitting predictions were compared between these two time series models to select the optimal one. The nosocomial infection data from January 2019 to June 2019 were used to test the predictive effect of the model.Results:A total of 379 477 cancer inpatients were studied, 3 170 of which acquired nosocomial infection and the incidence was 0.84% from 2013 to 2018. Additive Holt-Winters method exponential smoothing model was the better model with R2of 0.82. Using this model, the predicted value fitted well with observed value from January 2019 to June 2019, and the mean relative percentage error was 15.22%. Conclusion:Additive Holt-Winters method exponential smoothing model could be used to fit and predict the tendency of nosocomial infection among cancer patients, which can provide reference for surveillance of nosocomial infection in oncology hospitals.

Receptor-interacting protein (RIP) kinase 1 is involved in immune-mediated inflammatory diseases including ulcerative colitis (UC) by regulating necroptosis and inflammation. Our group previously identified TAK-632 (

Porcelain tooth technology is widely used in the treatment of oral diseases, but there are few reports on the possible occupational hazard factors in the process of porcelain tooth production. Porcelain teeth production will produced a large amount of silica dust and metal dust during the grinding process. The technical workers who have been engaged in this work for a long time are very prone to pneumoconiosis due to their poor personal protection awareness. This paper analyzed the clinical data of a pneumoconiosis patient engaged in porcelain tooth making, and analyzed the possible occupational hazard factors in the process of porcelain teeth production, so as to improve the understanding of relevant enterprises, technical workers and medical personnel on the disease and reduce the risk of porcelain teeth production workers suffering from pneumoconiosis.

Porcelain tooth technology is widely used in the treatment of oral diseases, but there are few reports on the possible occupational hazard factors in the process of porcelain tooth production. Porcelain teeth production will produced a large amount of silica dust and metal dust during the grinding process. The technical workers who have been engaged in this work for a long time are very prone to pneumoconiosis due to their poor personal protection awareness. This paper analyzed the clinical data of a pneumoconiosis patient engaged in porcelain tooth making, and analyzed the possible occupational hazard factors in the process of porcelain teeth production, so as to improve the understanding of relevant enterprises, technical workers and medical personnel on the disease and reduce the risk of porcelain teeth production workers suffering from pneumoconiosis.

To observe the migration of human amniotic mesenchymal stem cells (hAMSCs) labeled with PKH26 in the endometrium of rats intrauterine adhesion. hAMSCs were isolated, identified and labeled with PKH26 to detect the biological characteristics of the cells. Rat intrauterine adhesion models were established using mechanical and infective method and PKH26-labeled hAMSCs were transplanted through the tail vein. The distribution of PKH26 labeled hAMSCs in the endometrium of rats were observed with the fluorescence confocal microscope. The results showed that PKH26 stain had no significant effect on cell activity, cycle, apoptosis and so on. PKH26-labeled positive cells were mainly distributed in injured endometrium of rats. It shows that the PKH26 labeling technique is a safe and effective method for tracing the human amniotic mesenchymal stem cells in the treatment of intrauterine adhesions.

To study the biological function of DNAH2 (Homo sapiens dynein, axonemal, heavy chain 2) gene, we constructed human stable U2OS cell line of DNAH2 gene knockout through CRISPR/Cas9n double nick system. The A, B sgRNAs (Single guide RNA) and complementary strands were designed and synthesized. The double-stranded structures were formed during annealing, and connected with BbsⅠ cohesive ends-containing pX462 linear vector to construct the recombinant eukaryotic expression plasmids, including pX462-DNAH2-A and pX462-DNAH2-B. After the co-transfection of the two plasmids into U2OS cells, the addition of puromycin and limiting dilution method were used to obtain positive monoclonal cell line. Western blotting assay was then performed to detect the expression of DNAH2 protein, and PCR-sequencing technology was finally utilized to analyze the mutation feature. The results showed that A, B sgRNAs duplex was successfully inserted into pX462 vector, and DNAH2 protein was not expressed and DNAH2 gene suffered from the frame-shift mutation in U2OS-DNAH2-KO monoclonal cell line. These demonstrated that DNAH2 knockout U2OS stable cell line was successfully constructed through CRISPR/Cas9n double nick system, which providing a useful tool for the study of DNAH2 gene.