ObjectiveTo divide the muscle into different subzones according to different density ranges using the stratified analysis on the basis of myosteatosis， and to investigate the effect of muscle density changes on complications （Clavien-Dindo grade ≥Ⅲ） after orthotopic liver transplantation （OLT）. MethodsA retrospective analysis was performed for the medical records of 145 patients who underwent OLT in The First Hospital of Jilin University from May 2013 to September 2020， and with the plain CT scan images of the largest level of lumbar 3 vertebrae of each patient as the original data， Neusoft Fatanalysis software was used to measure related muscle parameters. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. The chi-square test or Fisher test was for comparison of categorical data between two groups. RIAS software was used to extract clinical features and perform analysis and modeling， and three machine learning models of logistic regression （LR）， support vector machine （SVM）， and random forest （RFC） were constructed. The receiver operating characteristic （ROC） curve， the calibration curve， and the decision curve were plotted for each model to calculate the area under the ROC curve （AUC）， sensitivity， specificity， precision， F1 score， and accuracy. ResultsThe three machine learning models of LR-C， SVM-C， and RFC-C were established based on the 7 clinical features before muscle stratification analysis， among which the RFC-C model had an AUC of 0.803， a sensitivity of 0.588， and a specificity of 0.778 in the test set. Among the models of LR-CS， SVM-CS， and RFC-CS established based on the 16 clinical features after muscle stratification analysis， the LR-CS and SVM-CS models had an AUC of 0.852 in the test set， with a sensitivity of 0.765 and 0.706， respectively， and a specificity of 0.889 and 0.926， respectively. Comparison of the AUC， sensitivity， specificity， precision， F1 score， and accuracy of each model in the test set before and after muscle stratification analysis showed that there were improvements in the parameters of the predictive model after muscle stratification analysis. Comparison of the decision curves and calibration curves of each predictive model showed that the LR-CS and SVM-CS models had good efficacy in predicting postoperative complications （Clavien-Dindo grade≥Ⅲ） in OLT patients. ConclusionOn the basis of myosteatosis， the division of the muscle into different subzones according to different densities using the stratified analysis has a certain value in predicting postoperative complications in patients with OLT.

BACKGROUND:In the treatment of skin trauma with active repair,tissue engineering techniques are needed to generate new tissue to replace necrotic tissue.Skin scaffolds have a good application prospect in the field of wound repair.Skin scaffolds need to present three-dimensional porous structures with certain mechanical strength to meet the needs of cell proliferation and division.However,the mechanical strength of the currently used gelatin-based biomaterials is weak and cannot meet the requirements of the use of skin scaffolds. OBJECTIVE:To study the 3D printing process used in the preparation of tissue engineering skin scaffolds by gelatin/oxidized nanocellulose composites,and focus on the relationship between the porosity and mechanical strength of the scaffolds prepared under different process parameters. METHODS:Oxidized nanocellulose whiskers at 10%concentration were extracted from Humulus scandens and then compounded with 5%gelatin to obtain gelatin/oxidized nanocellulose composites.The elastic modulus of gelatin and gelatin/oxidized nanocellulose composite was determined.Skin scaffolds were prepared by 3D printing extrusion molding using gelatin/oxidized nanocellulose composite as the base material.Mechanical and rheological properties of the composite were tested to determine extrusion molding parameters(filling gap 1.5-2.5 mm,uniform distribution of 0.1 mm;air pressure of 160-200 kPa),and the skin scaffold with a three-dimensional porous structure was prepared.The compressive performance of the skin scaffold was tested and compared with the finite element analysis results.The relationship between the filling gap and the porosity and mechanical strength of the scaffold was demonstrated. RESULTS AND CONCLUSION:(1)The elastic modulus of 5%gelatin was increased by 8.84 times by adding 10%oxidized nanocellulose whisker.A gel filament with a diameter of 1 mm was obtained by extrusion at the air pressure of 160 kPa.When the filling gap increased from 1.5 mm to 2.5 mm,the theoretical porosity of the scaffold increased from 33%to 60%,but the compressive strength decreased from 230 000 Pa to 95 000 Pa.(2)These findings showed that the skin scaffold with theoretical porosity of 50%and elastic modulus of 160 000 Pa was prepared by using 2 mm filling gap.The scaffold had a clear three-dimensional porous structure.

Perilla (Perilla frutescens L.) is an important edible-medicinal oil crop, with its seed containing 46%-58% oil. Of perilla seed oil, α-linolenic acid (C18:3) accounts for more than 60%. Lysophosphatidic acid acyltransferase (LPAT) is one of the key enzymes responsible for triacylglycerol assembly in plant seeds, controlling the metabolic flow from lysophosphatidic acid to phosphatidic acid. In this study, the LPAT2 gene from the developing seeds of perilla was cloned and designated as PfLPAT2. The expression profile of PfLPAT2 gene was examined in various tissues and different seed development stages of perilla (10, 20, 30, and 40 days after flowering, DAF) by quantitative real-time PCR (qRT-PCR). In order to detect the subcellular localization of PfLPAT2 protein, a fusion expression vector containing PfLPAT2 and GFP was constructed and transformed into Nicotiana benthamiana leaves by Agrobacterium-mediated infiltration. In order to explore the enzymatic activity and biological function of PfLPAT2 protein, an E. coli expression vector, a yeast expression vector and a constitutive plant overexpression vector were constructed and transformed into an E. coli mutant SM2-1, a wild-type Saccharomyces cerevisiae strain INVSc1, and a common tobacco (Nicotiana tabacum, variety: Sumsun NN, SNN), respectively. The results showed that the PfLPAT2 open reading frame (ORF) sequence was 1 155 bp in length, encoding 384 amino acid residues. Functional structure domain prediction showed that PfLPAT2 protein has a typical conserved domain of lysophosphatidic acid acyltransferase. qRT-PCR analysis indicated that PfLPAT2 gene was expressed in all tissues tested, with the peak level in seed of 20 DAF of perilla. Subcellular localization prediction showed that PfLPAT2 protein is localized in cytoplasm. Functional complementation assay of PfLPAT2 in E. coli LPAAT mutant (SM2-1) showed that PfLPAT2 could restore the lipid biosynthesis of SM2-1 cell membrane and possess LPAT enzyme activity. The total oil content in the PfLPAT2 transgenic yeast was significantly increased, and the content of each fatty acid component changed compared with that of the non-transgenic control strain. Particularly, oleic acid (C18:1) in the transgenic yeast significantly increased, indicating that PfLPAT2 has a higher substrate preference for C18:1. Importantly, total fatty acid content in the transgenic tobacco leaves increased by about 0.42 times compared to that of the controls, with the C18:1 content doubled. The increased total oil content and the altered fatty acid composition in transgenic tobacco lines demonstrated that the heterologous expression of PfLPAT2 could promote host oil biosynthesis and the accumulation of health-promoting fatty acids (C18:1 and C18:3). This study will provide a theoretical basis and genetic elements for in-depth analysis of the molecular regulation mechanism of perilla oil, especially the synthesis of unsaturated fatty acids, which is beneficial to the genetic improvement of oil quality of oil crops.
Acyltransferases ; Cloning, Molecular ; Escherichia coli/metabolism* ; Fatty Acids ; Perilla frutescens/metabolism* ; Plant Oils ; Plant Proteins/metabolism* ; Saccharomyces cerevisiae/metabolism* ; Seeds/chemistry* ; Tobacco/genetics*

ObjectiveTo observe the regulatory effect of Yuyetang on Ghrelin level in rats with type 2 diabetes mellitus （T2DM）-induced cognitive impairment （DCI） and explore the pathway in the prevention and treatment of DCI. MethodThe T2DM model was induced in rats by intraperitoneal injection of streptozotocin （STZ） combined with the high-fat and high-sugar diet （STZ）. The model rats were divided into model group， metformin group （200 mg·kg^-1）， and low-，medium-， and high-dose Yuyetang groups（4.575，9.15， 18.3 g·kg^-1）according to the blood glucose， with 10 rats in each group. A normal group was also set up. The rats were administered with corresponding drugs by gavage for 30 days， and the body weight and blood glucose of the rats in each group were observed and recorded. After drug intervention， the learning and memory abilities of rats were tested by the Morris water maze. After the test， the whole brains of rats were sampled for hematoxylin-eosin （HE） staining to observe the pathological changes in the hippocampal CA1 region， and the expression of Ghrelin in gastric tissues and hippocampal CA1 region was detected by immunohistochemistry. ResultCompared with the normal group ， the model group showed increased blood glucose（P<0.01），reduced body weight（P<0.01），prolonged escape latency（P<0.05，P<0.01）， shortened retention time and movement distance in the target area，decreased number of platform crossings（P<0.01）， abnormal morphology and structure of cells with disordered arrangement and reduced number in the hippocampal CA1 region， and decreased expression of Ghrelin in the serum，hippocampal CA1 region， and gastric tissues（P<0.05， P<0.01）. Compared with the model group， the medium- and high-dose Yuyetang groups showed increased body weight， while all Yuyetang groups showed reduced blood glucose（P<0.01）， shortened escape latency （P<0.05）， prolonged retention time and movement distance in the target area，increased platform crossings （P<0.05， P<0.01）， improved morphology and structure of cells， increased number of normal cell in the hippocampal CA1 region， and elevated Ghrelin levels in the serum， gastric tissues， and hippocampal CA1 region（P<0.05， P<0.01）. ConclusionYuyetang can effectively improve the cognitive ability of DCI rats， and its mechanism may be related to the regulation of Ghrelin levels in the serum， hippocampal CA1 region， and gastric tissues.

OBJECTIVE： To provide reference for the revision of China National Formulary for Children and make a sample for Chinese medical institutions to formulate their own formulary. METHODS： The suggestions on formulary revision and the formulation of formulary in medical institutions were put forward through comparing the selection principle， catalogues， chapters， drug items， formulary quantity and coincident drugs of World Health Organization （WHO） Model Formulary for Children （2010 edition） （WMFc）， British National Formulary （Children edition） （2016-2017 edition） （BNFc）， Chinese National Formulary （Children edition） （2013 edition） （CNFc）. RESULTS & CONCLUSIONS： The selection principle of WMFc was safe， effective and economical； that of BNFc was accurate and up-to-date， that of CNFc was safe， effective， economical and appropriate. The catalogues of three formularies included introduction， outline and separated section， but the contents or descriptions were different. For example， in separated section， WMFc was divided into 27 chapters， BNFc was divided into 16 chapters and CNFc was divided into 20 chapters. The chapters and catalogues of WMFc were classified according to ATC； those of BNFc were classified according to organ system and disease arrangement； those of CNFc were classified according to disease treatment system. 15 chapters of three formularies were the same， such as drug for nervous system diseases， drug for endocrine system and drug for respiratory system. The unique chapter of CNFc was “drugs for stomatological diseases”. The drug items of three formularies included drug name， indications， usage and dosage， contraindication， ADR， matters need attention， preparation and specifications， etc. Compared with CNFc， unique chapters of WMFc included ATC numbering， drug interaction， liver injury， etc.； those of BNFc included pharmacological action， interaction， allergy， etc. WMFc contained 271 drugs， CNFc 847 drugs and BNFc 955 drugs. Among them， there were 166 overlaps between WMFc and CNFc， 359 between BNFc and CNFc， 174 between WMFc and BNFc. There were 141 same drugs in the three formularies. When revising or formulating formulary， our country should not copy them mechanically， but should revise formulary according to our national conditions. It is necessary to refer to selection principles of WMFc and BNFc， update our children’s formulary in certain or real time， standardize drug access criteria of formulary， rationally increase the variety of drug and call on the relevant departments of the state to improve the standards for the formulation of drug instructions for children.

Objective? To explore effects of CoughAssist on sputum excretion efficacy among patients with intensive care unit-acquired weakness (ICU-AW). Methods? From January 2016 to December 2017, we selected 84 ICU-AW patients with mechanical ventilation of ICU in Hu'nan Provincial People's Hospital by convenience sampling. All of the patients were divided into control group and observation group with the random number table, 42 cases in each group. Two groups all received routine treatment and nursing. On this basis, observation group carried out CoughAssist therapy. We compared the sputum excretion effects, results of sputum smear/sputum culture, arterial blood gas indexes, respiratory mechanics indexes, the incidence of ventilator-associated pneumonia (VAP), time of mechanical ventilation, hospital days in ICU and the score of Medical Research Council (MRC) of patients between two groups. Results? There were no statistical difference in the volume of sputum excretion of patients in two groups before dividing groups (P＞0.05). One to three days after treatment, the volume of sputum excretion and number of effective sputum excretion patients in observation group were more than those in control group; the positive rate of sputum culture of patients in observation group was lower than that in control group; the differences were all statistical (P＜ 0.05). The partial pressure of oxygen (PaO2) of observation group was higher than that of control group; the partial pressure of carbon dioxide (PaCO2) of observation group was lower than that of control group; the differences were all statistical (P＜0.05). The pressure support/pressure control (PS/PC) and airway resistance of observation group were lower than those of control group;and the compliance and cough peak flow (PCF) were higher than those of control group with statistical differences (P＜ 0.01). The incidence of VAP, time of mechanical ventilation and hospital days in ICU of observation group were lower than those of control group (P＜0.05). There was no statistical difference in the scores of MRC between two groups (P＞0.05). Conclusions? The application of CoughAssist in ICU-AW patients with mechanical ventilation can improve the sputum excretion effects as well as respiratory function and shorten the time of mechanical ventilation and hospital days in ICU, and improve the clinical effects.

Objective? To explore the effects of structured hypothermia intervention program in elderly intestinal cancer patients with frailty. Methods? From November 2017 to July 2018, we selected 96 patients with surgery for intestinal cancer scored more than two points in Fried frailty assessment for the aged before surgery at the First Affiliated Hospital of Hu'nan Normal University & People's Hospital of Hu'nan Province by cluster sampling. All of the patients were divided into experimental group (n=48) and control group (n=48) with the method of random number table. Experimental group carried out the structured management model, formulated and implemented the structured hypothermia intervention program, while control group provided warm nursing based on nursing routine. We compared the respiration, blood pressure, heart rate and blood oxygen saturation before, during and after surgery as well as temperature, incidence of hypothermia and related indicators (extubation time, retention time in resuscitation room, urine volume and intraoperative blood loss) at each time points during surgery. Results were analyzed with the repeated measure variance analysis, t test and χ2 test. Results? There were interactions between time and groups in temperature, blood pressure, heart rate, respiration and blood oxygen saturation (P＜ 0.05). The fluctuation with time of the above vital signs in the experimental group was lower than that of control group. In experimental group, the incidence of hypothermia was lower than that in control group; the extubation time and retention time in resuscitation room were shorter than those in control group; the intraoperative blood loss was less than that in control group and the urine volume was more than that in control group;the differences were all statistically significant (P＜ 0.01). Conclusions? The application of structured hypothermia intervention program in elderly intestinal cancer patients with frailty can effectively maintain the stable intraoperative vital signs, shorten the time anesthesia resuscitation, reduce the incidences of hypothermia and related complications, standardize preventive measures for intraoperative hypothermia and guarantee patient intraoperative safety among elderly intestinal cancer patients with frailty.

Aging associated cognitive decline has been linked to dampened neural stem/progenitor cells (NSC/NPCs) activities manifested by decreased proliferation, reduced propensity to produce neurons, and increased differentiation into astrocytes. While gene transcription changes objectively reveal molecular alterations of cells undergoing various biological processes, the search for molecular mechanisms underlying aging of NSC/NPCs has been confronted by the enormous heterogeneity in cellular compositions of the brain and the complex cellular microenvironment where NSC/NPCs reside. Moreover, brain NSC/NPCs themselves are not a homogenous population, making it even more difficult to uncover NSC/NPC sub-type specific aging mechanisms. Here, using both population-based and single cell transcriptome analyses of young and aged mouse forebrain ependymal and subependymal regions and comprehensive "big-data" processing, we report that NSC/NPCs reside in a rather inflammatory environment in aged brain, which likely contributes to the differentiation bias towards astrocytes versus neurons. Moreover, single cell transcriptome analyses revealed that different aged NSC/NPC subpopulations, while all have reduced cell proliferation, use different gene transcription programs to regulate age-dependent decline in cell cycle. Interestingly, changes in cell proliferation capacity are not influenced by inflammatory cytokines, but likely result from cell intrinsic mechanisms. The Erk/Mapk pathway appears to be critically involved in regulating age-dependent changes in the capacity for NSC/NPCs to undergo clonal expansion. Together this study is the first example of using population and single cell based transcriptome analyses to unveil the molecular interplay between different NSC/NPCs and their microenvironment in the context of the aging brain.
Aging ; genetics ; Animals ; Astrocytes ; cytology ; metabolism ; Brain ; cytology ; metabolism ; Cell Differentiation ; genetics ; Cell Division ; genetics ; Cell Proliferation ; genetics ; Gene Expression Regulation ; genetics ; Mice ; Neural Stem Cells ; metabolism ; Single-Cell Analysis ; Stem Cells ; cytology ; metabolism ; Transcriptome ; genetics

By analyzing the present situation, advantages and challenges of the development of new nurses training in hospitals in the Massive Open Online Course (MOOC) era, it is suggested that the intelligent development of MOOC can not only meet the training needs of new nurses in the new situation, but also promote the application of MOOC in nursing inservice education. It is suggested that we should break the barriers, promote the sharing of high quality resources, improve the construction of curriculum system, strengthen the training supervision and assessment system, increase the policy support, strengthen the cultivation of MOOC technical personnel and technical innovation, and explore the "mixed teaching" model and other initiatives to promote MOOC science and its efficient development.

Objective To identify the activity of HCV IRES translation differences and identify the relationship between HCV IRES translation activity and ROS in different concentrations of ferric ammonium citrate ( FAC) in-duction.Methods 1 ) Expression plasmid pCI-Rluc-HCV IRES-Fluc was confirmed by endonuclease digestion as well as luciferase transient expression in Huh-7 cell;2) Controlled by dual-luciferase reporter assay, the differ-ent translation activity of HCV internal ribosomal entry site ( IRES ) was examined in a concentration of 50 μmol/L and 300μmol/L of FAC induction;ROS fluorescent staining method was used to detect the activity of ROS in Huh-7 cells, Western blot method was used to detect the protein expression changes of Nrf2 in Huh-7 cells;3) On the basis of the above experiments, 100 μmol/L DPI was added in 300 μmol/L FAC experimental group, to analyse the changes of HCV replication and ROS production after joining DPI.Results The generation of ROS and the activity of luciferase in the model group were significantly higher than that in the control group ( P<0.05 ) .FAC can enhance the expression of HCV IRES and increase the production of ROS , then causing Nrf2 expression in Huh-7 cell.However,after adding ROS inhibitor DPI, the above functions in Huh-7 cell were weakened.Conclusions The increase of HCV IRES expression induced by FAC is related to excessive ROS pro-duction induced by FAC in Huh-7 cells.