[Optimization of isolation and culture of Lin- Sca-1+ cardiac stem cells from newborn mice].

Cardiac stem cells (CSCs) transplantation has been recognized to be effective on the treatment of myocardial infarction (MI), but some techniques still need to be developed in the isolation and culture of CSCs, which is the key problem restricting the clinical application of CSCs. This study was focused on the isolation of Lin- (lineage-negative) Sca-1+ (stem cell antigen-1-positive) CSCs from newborn C57BL/6J mice (0-3 d) by mixed enzymatic-explant isolation in combination with immunomagnetic separation. The digesting time, digesting frequency, incubation temperature, stirring speed, centrifugation time and rotational speed were strictly controlled in the experiment. In order to increase the survival rate of CSCs, the medium changing time and manner were optimized in primary CSCs culture. The percentages of Sca-1+ cells in primary and passage cells were detected by flow cytometry and immunofluorescence staining. The results showed that: (1) the proportion of Lin- Sca-1+ cells within the collected cells could be as high as (85.03 ± 5.60)% after isolation and purification; (2) In vitro culture of Lin- Sca-1+ CSCs grew into spheres on the 5th day, and over the whole bottom of the dish on the 7th day. The growth curve showed that the cells were in logarithmic growth phase on the 3rd day; (3) Immunofluorescence staining data showed that the expression of Sca-1, the CSCs membrane-specific marker, was decreased after subculture, and flow cytometry data showed that the percentages of Sca-1+ cells were (71.82 ± 2.63)%, (58.38 ± 3.70)% and (46.19 ± 4.72)% in passage 1 (P1), P3, and P5 CSCs, respectively. The above results suggest that high purity of Lin- Sca-1+ CSCs can be obtained by enzymolysis combined with immunomagnetic separation method. Moreover, the CSCs culture system is stable. In our experiment, the Sca-1+ CSCs isolation and culture method has been successfully established, and it is simple, stable, effective and reliable. The method can provide a stable methodological basis for the treatment of MI by Lin- Sca-1+ CSCs transplantation.

Combined Transcriptomic and Proteomic Analysis of Myzus persicae, the Green Peach Aphid, Infected with Cucumber Mosaic Virus.

Aphids transmit CMV (cucumber mosaic virus) in a non-persistent manner. However, little is known about the mechanism of CMV transmission. In this study, an integrated analysis of the mRNA and protein was performed to identify important putative regulators involved in the transmission of CMV by aphids. At the level of transcription, a total of 20,550 genes (≥2-fold expression difference) were identified as being differentially expressed genes (DEGs) 24 h after healthy aphid transfer to infected tobacco plants using the RNA-seq approach. At the protein level, 744 proteins were classified as being differentially abundant between virus-treated and control M. persicae using iTRAQ (isobaric tags for relative and absolute quantitation) analysis. The combined mRNA and protein analysis enabled the identification of some viral putative regulators, such as cuticle proteins, ribosomal proteins, and cytochrome P450 enzymes. The results show that most of the key putative regulators were highly accumulated at the protein level. Based on those findings, we can speculate that the process by which aphids spread CMV is mainly related to post-translational regulation rather than transcription.

New Insights into the Mechanism of Enhanced Performance of Li[Ni0.8Co0.1Mn0.1]O2 with a Polyacrylic Acid-Modified Binder.

A binder is an important component in lithium-ion batteries and plays a significant role in maintaining the properties of active substances. Most studies in the field of binders have only focussed on physical properties such as bonding performance. Here, a polyacrylic acid-modified binder was designed and adapted to Li[Ni0.8Co0.1Mn0.1]O2, which enhanced the electrochemical stability of Li[Ni0.8Co0.1Mn0.1]O2 from 30.2 to 66.6% (300 cycles at 1 C). We for the first time discovered that this was caused by a chemical reaction between polyacrylic acid and the residual lithium on the surface during the cycling, which formed a lithium propionic acid coating layer and maintained the stability of the layered structure.

Biofilm Production Ability, Virulence and Antimicrobial Resistance Genes in Staphylococcus aureus from Various Veterinary Hospitals.

: Staphylococcus aureus (S. aureus) is one of the most clinically important zoonotic pathogens, but an understanding of the prevalence, biofilm formulation ability, virulence, and antimicrobial resistance genes of S. aureus from veterinary hospitals is lacking. By characterizing S. aureus in different origins of veterinary hospitals in Guangzhou, China, in 2019, we identified with the presence of S. aureus in pets (17.1%), veterinarians (31.7%), airborne dust (19.1%), environmental surfaces (4.3%), and medical device surfaces (10.8%). Multilocus sequence typing (MLST) and Staphylococcus protein A (spa) typing analyses demonstrated methicillin-sensitive S. aureus (MSSA) ST398-t571, MSSA ST188-t189, and methicillin-resistant S. aureus (MRSA) ST59-t437 were the most prevalent lineage. S. aureus with similar pulsed-field gel electrophoresis (PFGE) types distributed widely in different kinds of samples. The crystal violet straining assays revealed 100% (3/3) of MRSA ST59 and 81.8% (9/11) of MSSA ST188 showed strong biofilm formulation ability, whereas other STs (ST1, ST5, ST7, ST15, ST88, ST398, ST3154 and ST5353) showed weak biofilm production ability. Polymerase chain reaction (PCR) confirmed the most prevalent leucocidin, staphylococcal enterotoxins, ica operon, and adhesion genes were lukD-lukE (49.0%), sec-sel (15.7%), icaA-icaB-icaC-icaR (100.0%), and fnbB-cidA-fib-ebps-eno (100.0%), respectively. Our study showed that the isolates with strong biofilm production ability had a higher prevalence in clfA, clfB, fnbA and sdrC genes compared to the isolates with weak biofilm production ability. Furthermore, 2 ST1-MRSA isolates with tst gene and 1 ST88-MSSA isolate with lukS/F-PV gene were detected. In conclusion, the clonal dissemination of S. aureus of different origins in veterinary hospitals may have occurred; the biofilm production capacity of S. aureus is strongly correlated with ST types; some adhesion genes such as clfA, clfB, fnbA, and sdrC may pose an influence on biofilm production ability and the emergence of lukS/F-PV and tst genes in S. aureus from veterinary hospitals should raise our vigilance.

[Frequency up-conversion properties of Er3+ /Yb3+ co-doped zinc oxide powders].

Er3+ /Yb3+ co-doped ZnO powders were prepared by the high temperature sintering method with starting composition of (mol%) 95ZnF2-4. 8Yb2 O3-0. 2Er2 O3. Microstructure analysis by X-ray diffraction (XRD) showed that the sample consists of two phases, i. e. ZnO and YbF3, which verified that the ZnF2 was oxidized during the high temperatue sintering Composition analysis by scanning electron microscope (SEM) and spectroscopic measurements showed that the Er3+ and Yb3+ ons were successfully used in doping the lattice of ZnO, but most of Yb3+ ions were in the YbF3 phase. These results indicated that the up-conversion luminescence was emitted from ZnO, not from YbF3. Under the excitation of 980 nm diodelaser, four strong up-conversion emissions peaks centered at 658, 538, 522 and 409 nm, corresponding to the transitions 4F9/2 --> 4I15/2, 4S3/2 --> 4I15/2, 2 H11/2 --> 4I15/2 and 2 H9/2 --> I15/2, respectively, were observed. Especially, a strong red up-conversion emission was observed, which is different from that the green up-converted luminescence is dominated in glass and ceramics. Three important cross energy transfer (CRET) processes between Er3+ ions played an important role for this. Under 488 nm Ar+ laser excitation, intense violet (409 nm), weak blue (466, 450 nm) and ultraviolet (379 nm) up-conversion luminescence originating from the transitions 2 H9/2 --> 4I15/2, 2P3/2 --> 4I11/2, 4 F3/2 /4 F5/2 --> 4I15/ 2 and 4G11/2 --> 4 I15/2, respectively, were obtained. The dependence of up-conversion intensities on excitation power indicated that two-photon absorption processes were responsible for the violet luminescence under 488 nm excitation, and the violet up-converted luminescence was achieved through the forward and back energy transfer between Er3+ and Yb3+ ions. Our results show that ZnO as a host material has the potential applications in the up-conversion red phosphors and ultraviolet laser materials.

ClCRY2 facilitates floral transition in Chrysanthemum lavandulifolium by affecting the transcription of circadian clock-related genes under short-day photoperiods.

Plants sense photoperiod signals to confirm the optimal flowering time. Previous studies have shown that Cryptochrome2 (CRY2) functions to promote floral transition in the long-day plant (LDP) Arabidopsis; however, the function and molecular mechanism by which CRY2 regulates floral transition in short-day plants (SDPs) is still unclear. In this study, we identified a CRY2 homologous gene, ClCRY2, from Chrysanthemum lavandulifolium, a typical SDP. The morphological changes in the C. lavandulifolium shoot apex and ClFTs expression analysis under SD conditions showed that adult C. lavandulifolium completed the developmental transition from vegetative growth to reproductive growth after eight SDs. Meanwhile, ClCRY2 mRNA exhibited an increasing trend from 0 to 8 d of SD treatment. ClCRY2 overexpression in wild-type (WT) Arabidopsis and C. lavandulifolium resulted in early flowering. The transcript levels of the CONSTANS-like (COL) genes ClCOL1, ClCOL4, and ClCOL5, and FLOWERING LOCUS T (FT) homologous gene ClFT1 were upregulated in ClCRY2 overexpression (ClCRY2-OE) C. lavandulifolium under SD conditions. The transcript levels of some circadian clock-related genes, including PSEUDO-REPONSE REGULATOR 5 (PRR5), ZEITLUPE (ZTL), FLAVIN-BINDING KELCH REPEAT F-BOX 1 (FKF1), and GIGANTEA (GI-1 and GI-2), were upregulated in ClCRY2-OE C. lavandulifolium, while the expression levels of other circadian clock-related genes, such as EARLY FLOWERING 3 (ELF3), ELF4, LATE ELONGATED HYPOCOTYL (LHY), PRR73, and REVEILLE8 (RVE8), were downregulated in ClCRY2-OE C. lavandulifolium under SD conditions. Taken together, the results suggest that ClCRY2 promotes floral transition by fine-tuning the expression of circadian clock-related gene, ClCOLs and ClFT1 in C. lavandulifolium under SD conditions.

[Fluorescence spectra and analysis of chicken liver GST].

Glutathione transferases (GST; EC2. 5. 1. 18) is an important detoxification enzyme which catalyze the conjugation of glutathione to a large variety of endogenous and exogenous electrophilic compounds to protect the functions of body. In the present paper, three dimensional fluorescence spectra were obtained, through which the authors could identify the fluorescence spectra of peptide bond, Tyr and Trp residue. The authors compared aromatic amino-acid residue fluorescence spectra in GST with dissociative to know red or blue shift of the fluorescence peak. The authors also studied the peptide bond and Trp residue fluorescence spectra at various pH, which suggested the change in GST surface and surface hydrophobicity, and the microenviroment change of aromatic amino-acid residue in enzyme.

Comparative analyses of light-induced anthocyanin accumulation and gene expression between the ray florets and leaves in chrysanthemum.

Light is one of the key environmental factors that affect anthocyanin biosynthesis. However, the underlying molecular mechanism remains unclear, and many problems regarding phenotypic change and corresponding gene regulation have not been solved. In the present study, comparative analyses of light-induced anthocyanin accumulation and gene expression between the ray florets and leaves were performed in Chrysanthemum × morifolium 'Purple Reagan'. After contrasting the variations in the flower color phenotype and relative pigment content, as well as expression patterns of structural and regulator genes responsible for anthocyanin biosynthesis and photoreceptor between different plant organs under light and dark conditions, we concluded that (1) both the capitulum and foliage are key organs responding to light for chrysanthemum coloration; (2) compared with flavones, shading makes a greater decrease on the anthocyanins accumulation; (3) most of the structural and regulatory genes in the light-induced anthocyanin pathway specifically express in the ray florets; and (4) CmCHS, CmF3H, CmF3'H, CmANS, CmDFR, Cm3GT, CmMYB5-1, CmMYB6, CmMYB7-1, CmbHLH24, CmCOP1 and CmHY5 are key genes for light-induced anthocyanin biosynthesis in chrysanthemum ray florets, while on the transcriptional level, the expressions of CmPHYA, CmPHYB, CmCRY1a, CmCRY1b and CmCRY2 are insignificantly changed. Moreover, the inferred comprehensive effect of multiple signals on the accumulation of anthocyanins and transmission channel of light signal that exist between the leaves and ray florets were further discussed. These results further our understanding of the relationship between the gene expression and light-induced anthocyanin biosynthesis, and lay foundations for the promotion of the molecular breeding of novel flower colors in chrysanthemums.

[Luminescence properties and up-conversion mechanism of Er3+ -Yb3+ -Tm3+ co-doped CdF2: PbF2-based glass].

Under 980 nm excitation, we obtained five intense up-conversion luminescence bands in Er3+ -Yb3+ -Tm3+ co-doped CdF2: PbF2-based glass as follows: infrared (800 nm), red (645 nm), green (545 nm and 525 nm), blue (480 nm ) and violet (407 nm) up-conversion luminescence. With the addition of Tm3+ , the intensity of 480 nm blue luminescence increased notably compared with that of co-doped Yb3+ -Er3+ sample, which attributed to Tm3+ special energy level structure; logI-logP plot of the luminescence intensity versus pump power shows that 480 nm luminescence emission is a two-photon excitation process induced by the cooperative up-conversion between two Yb3+ ions and the slope of logI-logP plot decreases gradually with increasing pump power and shows a tendency to bend down. The up-conversion mechanism is analyzed in detail under 980 nm semiconductor laser excitation; the dependence of 480 nm up-conversion luminescence intensity on the pump power under the steady states is discussed by using rate equation and the results agree well with the experiments.

Lanthanide doping-facilitated growth of ultrasmall monodisperse Ba2LaF7 nanocrystals with excellent photoluminescence.

Lanthanide doping not only works as sensitizer and activator, but also plays an important role to facilitate the growth of nanocrystal and to control the size, shape, and property of nanocrystals. Here, reported was the synthesis of monodisperse Ba(2)LaF(7) nanocrystals with the size of sub-10nm through a solvothermal method. We found the dopants of Ho(3+), Er(3+), or Yb(3+) facilitated the growth of Ba(2)LaF(7) nanocrystals obviously to a certain size within a shorter reaction time. Similar phenomenon can also be observed in the synthesis of LaF(3) nanocrystals. We find that Ln(3+) (e.g., Ho(3+), Er(3+), or Yb(3+)) with smaller radius can reduce the nucleation energy and lead to heterogeneous nucleation, which favors the growth of Ba(2)LaF(7) nanocrystals obviously. In addition, intense upconversion emission can be observed from Ln(3+)-doped Ba(2)LaF(7) nanocrystals under the 980 nm laser excitation, providing great potential application in biological imaging. Especially, Ba(2)LaF(7):Yb/Er (20/1 mol%) nanocrystals present more intense upconversion emission than α-NaYF(4):Yb/Er (20/1 mol%) nanocrystals under the same conditions.

[Effects of exogenous gamma-aminobutyric acid on polyamine metabolism of melon seedlings under hypoxia stress].

Taking melon cultivar 'Xiyu No. 1 ' as test material, a hydroponic experiment was conducted to investigate the effects of exogenous gamma-aminobutyric acid (GABA) on the seedlings polyamine metabolism under hypoxia stress. Compared with the control in normoxic treatment, the seedlings under hypoxia stress had significantly higher glutamic acid decarboxylase (GAD) activity and GABA content, and their polyamine synthesis enzymes activities all enhanced significantly, which led to a marked increase of polyamines contents. Meanwhile, the seedlings leaf- and root diamine oxidase (DAO) and polyamine oxidase (PAO) activities also had a significant increase. The increment of root arginine decarboxylase (ADC) activity was higher, which induced a higher content of free putrescine (Put) in roots, while the increment of leaf ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) activities were higher, inducing a higher content of free spermidine (Spd) in leaves. The PBs-extractable DAO and PAO activies in roots were significantly lower than those in leaves, but the cell wall-bound PAO activity was in adverse. Under hypoxia stress, the addition of exogenous GABA increased the leaf- and root GABA and glutamic acid contents and decreased the GAD activity significantly. The increase of arginine, ornithine, and methionine contents promoted the activities of polyamines synthesis enzymes, which led to the significant increase of polyamines contents and the significant decrease of DAO and PAO activities.

[Effects of organic manure on wheat growth under lead stress].

A pot experiment was conducted to study the effects of organic manure on the wheat growth under different levels of lead stress. With increasing lead stress level, whether fertilization or not, the plant height, shoot dry mass, adventitious root number, root total length, root dry mass, root activity, root total and active absorbing area, and root SOD and POD activities decreased, and root MDA content presented an increasing trend. The decrement of the above-mentioned parameters differed with fertilization treatments. Applying organic manure mitigated the impact of lead stress on wheat growth to some extent, delayed the senescence of wheat roots, and promoted root development and growth, ultimately leading to the increase of wheat yield and the decrease of lead content in grain.