A NEW METHOD OF ISOLATING KUPFFER CELLS FROM BIOPSY TISSUE OF RAT LIVER

The isolation of a high yield and purity of Kupffer cells has been reported in detail.1 This paper reports into the research about isolation Kupffer cells from biopsy tissue of liver. This method includes 5 important steps: (1) take fresh liver tissue, and mince with scissors. (2) spin at low speed to wash off red blood cells. (3) digest in collagenase for suitable time. (4) isolate Kupffer cells on a percoll density gradient. (5) cell charaterization was observed by N.S.E stain and peroxidatic activity with lumino-meter measurement and phagocytosis with latex beads.2.3

Extracellular vesicles in Alzheimer’s disease:from pathology to therapeutic approaches

Alzheimer’s disease is a progressive and fatal neurodegenerative disorder that starts many years before the onset of cognitive symptoms.Identifying novel biomarkers for Alzheimer’s disease has the potential for patient risk stratification,early diagnosis,and disease monitoring in response to therapy.A novel class of biomarkers is extracellular vesicles given their sensitivity and specificity to specific diseases.In addition,extracellular vesicles can be used as novel biological therapeutics given their ability to efficiently and functionally deliver therapeutic cargo.This is critical given the huge unmet need for novel treatment strategies for Alzheimer’s disease.This review summarizes and discusses the most recent findings in this field.

Vibration Isolation Characteristics of Impedance-balanced Ship Equipment Foundation under Unbalanced Excitation

A new type of impedance-balanced ship equipment foundation structure based on the principle of impedance balancing using a“discontinuous panel-vibration isolation liquid layer-foundation structure”is proposed to solve the problem of poor low-frequency vibration isolation of the foundation under unbalanced excitation of shipboard equipment.Based on the finite element method,the influence of characteristic parameters of the foundation panel structure on its vibration reduction characteristics under unbalanced excitation is explored.The results show that the vibration isolation level of the impedance-balanced foundation is 10 dB higher than the traditional foundation in the low-frequency band of 10-500 Hz when subjected to combined excitation of concentrated force and moment.Increasing the thickness of the impedance-balanced foundation panel can enhance the isolation effect.Increasing the number of sub-panels can effectively reduce the vibration response of the foundation panel and enhance the isolation performance of the foundation.The connection stiffness between sub-panels has a small effect on the isolation performance of the foundation.

Analysis of All-Carbon Brick Bottom and Ceramic Cup Synthetic Hearth Bottom

One of the bottlenecks of the blast furnace （BF） campaign is the life length of hearth bottom. The basic reason for the erosion of hearth bottom is its direct contact with hot metal. According to the theory of heat transfer, models of BF hearth bottom are built based on the actual examples using software and VC language, and the calculated results are in good agreement with the data of BF dissection after blowing out. The temperature distribution and the capability of the resistance to erosion for different structures of hearth bottom are analyzed, especially the two prevalent kinds of hearth bottom arrangements called ＂the method of heat transfer＂ for all-carbon brick bottom and ＂the method of heat isolation＂ for ceramic synthetic hearth bottom. Features of the two kinds of hearth bottoms are analyzed. Also the different ways of protecting the hearth bottom are clarified, according to some actual examples. After that, the same essence of prolonging life, and the fact that the existence of a ＂protective skull＂ with low thermal conductivity between the hot metal and brick layers is of utmost importance are shown.

Isolation of Chloroplasts from Marine Microalga Isochrysis galbana Parke for Their Lipid Composition Analysis

Marine microalga Isochrysis galbana is an important feed species with a high nutritional value.Different from other uni-cellular algae,its cell contains two chloroplasts which are the major sites for lipid synthesis.Here,we optimized a chloroplast isola-tion approach suitable for the isolation of I.galbana chloroplasts and determined the purity and integrity of the isolated chloroplasts through microscopic observations and enzyme activity assay.The chloroplast lipids were analyzed with a ultrahigh-performance li-quid chromatography-Q Exactive Orbitrap-mass spectrometry.This newly developed isolation approach is simple and reliable to isolate chloroplasts with high integrity and purity.The average yield of intact chloroplasts was 15.3%±0.1%.Glycolipids and acyl-glycerols were the main chloroplast lipids.Glycolipids accounted for 56.6%of chloroplast lipid.Digalactosyldiacylglycerol(DGDG),monogalactosyldiacylglycerol(MGDG)and sulfoquinovosyldiacylglycerol(SQDG)were the main glyceroglycolipids.The fatty acyl R1/R2 were mostly 18:4/16:1,18:3/16:1 and 18:4/18:5 in DGDGs,14:0/18:4,18:4/18:5,18:4/18:4 and 18:3/18:4 in MGDGs and 16:0/14:0,16:0/18:3,and 18:4/18:3 in SQDGs.In addition,diacylglycerol(DAG)was the most abundant acylglycerols;the content of 22:6/18:4-DAG was the highest.There was a little amount of glycosphingolipid(GSL)in chloroplast.Digalactosylmonoglyceride(DGMG),monogalactosylmonoglyceride(MGMG),sulfoquinovosylmonoacylglycerol(SQMG),monoglyceride(MAG),phospholi-pids(PLs),ceramide(Cer)and betaine lipids were nearly undetectable in chloroplast.The fatty acid proportions of DGDGs,MGDGs,SQDGs,DAGs,triglycerides(TAGs)and GSLs were either higher or lower than or similar to those of whole-cell.Collectively,our isolation approach is applicable to many aspects of chloroplast biology,and may offer a reference for the isolation of chloroplasts from other marine microalgae.

Effective methods for isolation and purification of extracellular vesicles from plants

Plant extracellular vesicles(EVs)play critical roles in the cross-kingdom trafficking of molecules from hosts to interacting microbes,most notably in plant defense responses.However,the isolation of pure,intact EVs from plants remains challenging.A variety of methods have been utilized to isolate plant EVs from apoplastic washing fluid(AWF).Here,we compare published plant EV isolation methods,and provide our recommended method for the isolation and purification of plant EVs.This method includes a detailed protocol for clean AWF collection from Arabidopsis thaliana leaves,followed by EV isolation via differential centrifugation.To further separate and purify specific subclasses of EVs from heterogeneous vesicle populations,density gradient ultracentrifugation and immunoaffinity capture are then utilized.We found that immunoaffinity capture is the most precise method for specific EV subclass isolation when suitable specific EV biomarkers and their corresponding antibodies are available.Overall,this study provides a guide for the selection and optimization of EV isolation methods for desired downstream applications.

Rapid and efficient method for isolating plant high molecular weight(HMW)DNA with high purity

PREPARATION of HMW DNA (Megabase-size) is the basis for construction of genomic library with large DNA inserts such as bacterial artificial chromosome (BAC) and yeast artificial chromosome (YAC), and for long-range physical mapping. It can also be used for the macro-study of repeat sequences. Since HMW DNA during preparation is inclined to be sheared physically and digested by internal nucleases, it is very difficult to prepare the HMW DNA. Initially, plant HMW DNA was prepared by embedding protoplasts in the low melting-point (LMP) agarose; however, it had several disadvantages: (ⅰ) Culture of protoplasts was time-consuming, costly and tedious. ( ⅱ ) It was only used successfully for limited