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2024
Volume: 33 Issue 3
12 Article(s)
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Plant Growth-promoting Rhizobacteria: Mechanisms and Perspectives
LI Xiaochun, SONG Kai, CHEN Bo, JIANG Lian, and HE Yawen
The bacteria that live in the rhizosphere and promote the growth of host plants are called plant growth-promoting rhizobacteria (PGPR). PGPR application helps to reduce the use of pesticides and fertilizers and contributes to the sustainable development of agriculture. Over the past 20 years, significant progresses havThe bacteria that live in the rhizosphere and promote the growth of host plants are called plant growth-promoting rhizobacteria (PGPR). PGPR application helps to reduce the use of pesticides and fertilizers and contributes to the sustainable development of agriculture. Over the past 20 years, significant progresses have been made in both theoretical and applied re- searches of PGPR. This review aims to summarize the mechanisms underlying PGPR growth promotion and disease resistance, including improving nutrient supply and uptake, producing phytohormone and signal molecules, synthesizing volatile organic compounds, and secreting antibiotics. The future research direction of PGPR in both basic and application aspects is also dis- cussed, hoping to provide theoretical reference for future PGPR related research..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 193 (2024)
Research on the Role of Folic Acid in the Development of AnimalEmbryonic Organs
ZENG Xin, CAI Xiaoyan, and LI Fang
Folic acid, also known as vitamin B9, plays a key role in DNA synthesis, RNA transcription, homocysteine to methionine and cellular metabolism. Embryonic development is regulated by both environmental and genetic factors. Folic acid isan important nutrient for the normal development of animal embryonic organs. MaternaFolic acid, also known as vitamin B9, plays a key role in DNA synthesis, RNA transcription, homocysteine to methionine and cellular metabolism. Embryonic development is regulated by both environmental and genetic factors. Folic acid isan important nutrient for the normal development of animal embryonic organs. Maternal folic acid supplementation can reducethe risk of congenital organ dysplasia, such as heart, eyes, neural tube and kidney development abnormalities. In this study, weelucidates the involvement of folic acid transport, folic acid metabolism and the relationship between folic acid supplementationand epigenetic modifications in early embryonic organ development. In addition, we analyzed the effect of folic acid deficiencyon the development of animal embryonic organs and the importance of folic acid supplementation during pregnancy, providing atheoretical basis for the prevention and treatment of abnormal embryonic organ development in the future..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 201 (2024)
Segmentation and Quantification of Zebrafish Hindbrain Vessel ConfocalImages Based on DE-3D U-Net
ZHENG Wenhu, LI Hui, CHEN Chong, and WANG Linbo
The choroid plexues in the zebrafish hindbrain controls the exchange of substances between brain and its vessels,and the vascular abnormalities around the choroid plexues can lead to associated cerebrovascular diseases. We first performedserial imaging of the major blood vessels in the hindbrain region of zebrafish, anThe choroid plexues in the zebrafish hindbrain controls the exchange of substances between brain and its vessels,and the vascular abnormalities around the choroid plexues can lead to associated cerebrovascular diseases. We first performedserial imaging of the major blood vessels in the hindbrain region of zebrafish, and obtained a series of images of zebrafish between 43 hours and 63 hours after fertilization using confocal microscopy to photograph embryos under normal conditions andafter cylindrospermopsin treatment. Afterwards, the DE-3D U-Net was developed for the segmentation of the hindbrain images.The segmentation results showed that the class pixel accuracy of DE-3D U-Net could reach to 86.67%, 93.18% and 83.74% forthe three main vessels and the segmentation results were used to plot vascularization curves. The quantitative results showed thatthe radii of the three vessels showed a reduction of 18% after treatment with high concentrations of cylindrospermopsin, and theangle of the mesencephalic vein showed a reduction of 30° to 50°. This study proposed a neural network for segmentation ofhindbrain vessels of zebrafish, and it provided a basis for research on the development of zebrafish choroid plexsus..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 209 (2024)
Optimization Design of Sequencing Module for Domestic Genetic Analyzer
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A domestic 24 channels genetic analyzer combined with a domestic 36 cm capillary arrays were used to solve theproblems of fluorescence signal correction, running voltage, and migration correction during sequencing on the genetic analyzerbased on non-gel sieving capillary electrophoresis technology. Spectral calibrationA domestic 24 channels genetic analyzer combined with a domestic 36 cm capillary arrays were used to solve theproblems of fluorescence signal correction, running voltage, and migration correction during sequencing on the genetic analyzerbased on non-gel sieving capillary electrophoresis technology. Spectral calibration model and appropriate baseline noise valueswere obtained under the condition that the original spectral fluorescence signal of the analyzer was used as a regulation basis. Astandard curve was plotted by combining an running voltage and a base spacing value, while a clear range length was also calculated by a sequencing analysis software, so as to determine the optimal running voltage and base spacing. In addition, a migration offset value was further calculated and a linear model was eventually established between the base size and the migrationtime, which achieved an accurate identification of bases. It was reported that with the appropriate spectral calibration model andbaseline noise threshold limitation, the strongest ability to detect the longest clear range length, that is 561 bp, was found in theanalyzer when the running voltage was set to 10 kV and the base spacing was 13.05 frames. By compensating for migration corrections, a linear model was established between the base size and the migration time. Among them, R2 values of bases G, A,T, and C increased from 0.992 7, 0.992 7, 0.994 5, and 0.987 9 to 0.999 6, 0.999 8, 0.999 6, and 0.999 7, respectively. Aftercorrection, all fluorescent bases were accurately labeled without any omissions or errors, clear range length extended to 621 bp.This study could better guide the optimization and design of a sequencing module of the domestic genetic analyzer, making theDNA base recognition function of the analyzer more efficient and accurate..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 217 (2024)
Expression, Purification, and Preliminary Crystallographic Study ofFerritin from Spiroplasma eriocheiris
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Ferritin is a class of proteins ubiquitously found in various types of nucleated organisms and involved in the ironhomeostasis regulation of body. Ferritin can self-assemble to form a hollow nanocage of about 14 nm in size and is often usedas carrier for drug delivery and vaccine construction. The crystal structure of Ferritin is a class of proteins ubiquitously found in various types of nucleated organisms and involved in the ironhomeostasis regulation of body. Ferritin can self-assemble to form a hollow nanocage of about 14 nm in size and is often usedas carrier for drug delivery and vaccine construction. The crystal structure of Mycoplasma ferritin shows atypical ferroxidasecentres and new pathway for Fe2+ uptake, which are structurally different from those of other species. Not much work has beenreported for Spiroplasma ferritin. Here, our studies show that Spiroplasma ferritin has low sequence similarity with other ferri tins including Mycoplasma ferritins. Spiroplasma eriocheiris Ferritin (SeFer) was recombinant expressed in E. coli in solubleform after IPTG (isopropyl-beta-D-thiogalactopyranoside) induction. High-purity SeFer protein was obtained after purificationby DEAE (diethylaminoethyl) weak anion exchange chromatography and gel filtration. Crystal screen of SeFer was performedby sitting drop vapor diffusion at 4℃ and 18℃. SeFer crystals grew in several screen conditions. The best crystal diffracted to2.90 ? by synchrotron radiation source and it belonged to the I432 space group. The X-ray crystallography data collected herecan be used for further structure determination. Native-PAGE, dynamic light scattering, and the position of peaks of SeFer inmolecular sieve showed that the molecular weight of SeFer is about 400.0 kD, and the diameter of SeFer was about 15 nm, bothof them are close to the theoretical value. The structural model of SeFer was predicted by using Alphafold 2. The predicted mod el shows that SeFer has a regular icosahedral overall structure, which is similar to other ferritins. However, SeFer model showsthat the key residues of ferroxidase center differ from those of mycoplasma ferritin. Our results not only provide a basis forstudying the crystal structure of SeFer, but also provide new candidates for vaccine vectors..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 227 (2024)
A Study on Mechanism of ASM in the Treatment of ALI Based onNetwork Pharmacology and Molecular Docking
FENG Ling, HUANG Zhaoxia, LI Junlan, DONG Zhengping, and ZHANG Yongqin
Based on network pharmacology and molecular docking technology, the mechanism of radix angelicae sinensis-radix salviae miltiorrhizae drug pair (ASM) in the treatment of acute liver injury (ALI) was studied and verified by experiment. Inthis study, network pharmacology and molecular docking methods were used to screen Based on network pharmacology and molecular docking technology, the mechanism of radix angelicae sinensis-radix salviae miltiorrhizae drug pair (ASM) in the treatment of acute liver injury (ALI) was studied and verified by experiment. Inthis study, network pharmacology and molecular docking methods were used to screen the core targets of ASM in the treatmentof ALI, and the core targets were enriched by analysis, and protein interaction network was constructed. The core target was molecular docked with the active components of ASM, and the molecular docking was verified by constructing ALI rat model anddetecting the expression of relevant serum biochemical indexes and liver proteins. The results showed that 67 active ingredients were obtained from ASM, of which 75 targets interacted with acute liver injury. Protein-protein interactions (PPI) networkanalysis showed that TP53, CASP3, JUN, STAT3, AKT1, VEGFA, TNF, IL-6, MMP9 and PTGS2 may be the key targets ofASM in the treatment of ALI. The core targets of GO and KEGG signaling pathway enrichment analysis were mainly concentrated in hepatitis B signaling pathway, lipid and atherosclerosis, AGE-RAGE, and apoptosis signaling pathways. Moleculardocking results showed that the core targets (TP53 and CASP3) had good binding sites with β-sitosterol, baicalin and cryptotanshinone. The results of in vivo experiments showed that ASM could reduce the expression levels of ALT, AST and MDA in serum of CCl4-induced model rats, and increase the activity of SOD, CAT and GSH in liver, thereby alleviating the CCl4-inducedoxidative stress. The ASM could up-regulate BCL2 and down-regulate the expression levels of BAX, CASP3 and TP53. Theresults of this study indicate that ASM can effectively ameliorate ALI, which may be achieved through β-sitosterol, baicalin andcryptotanshinone and other chemical components, acting on key targets such as TP53 and CASP3, and through multiple signaling pathways such as p53 signaling pathway and apoptosis pathway. This study provides theoretical and experimental basis forclinical ASM treatment of ALI, and provides new ideas and methods for further research and development of traditional Chinese medicine..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 243 (2024)
Evolutional and Functional Analysis of Plant GYF Domains
LU Xun, LU Jianhua, PENG Shuming, XIA Qingzhu, PENG Bo, LI Benben, and ZHANG Xiaobo
According to the extensive distribution of GYF domain proteins and the diversity of its functions, it is necessary tostudy GYF domain proteins further and conduct evolutionary analysis. Since AtEXA1 is the most well-studied GYF domain proteinin plants, this study mainly focuses on the proteins similar to AtEXA1. AnalysAccording to the extensive distribution of GYF domain proteins and the diversity of its functions, it is necessary tostudy GYF domain proteins further and conduct evolutionary analysis. Since AtEXA1 is the most well-studied GYF domain proteinin plants, this study mainly focuses on the proteins similar to AtEXA1. Analysis of 8 053 GYF domain proteins in eukaryotes ranging from unicellular lower fungi to higher organisms showed that AtEXA1-like proteins are mainly found in plant species, whileGYF proteins from animals have low similarity to AtEXA1. It was also found that AtEXA1-like proteins are present in almost allagricultural crops. This study demonstrates the conservation of all GYF domain proteins, initially deconstructs the evolution of GYFdomain proteins, and provides implications for subsequent studies on the functional characteristics of GYF domain proteins..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 253 (2024)
Catechin Alleviates Apoptosis, Inflammation and Oxidative Damageof Rat Cardiomyocytes Induced by Lipopolysaccharide by Inhibitingp38 MAPK Phosphorylation
XING Jianhua, SUN Lulu, and LI Zhexian
To investigate the effects of catechins on oxidative damage, inflammation and apoptosis of rat cardiomyocytes(H9C2) induced by lipopolysaccharide (LPS), as well as the regulation of p38 mitogen-activated protein kinase (MAPK) sig naling pathway, H9C2 cells were cultured in vitro and divided into the control group whichTo investigate the effects of catechins on oxidative damage, inflammation and apoptosis of rat cardiomyocytes(H9C2) induced by lipopolysaccharide (LPS), as well as the regulation of p38 mitogen-activated protein kinase (MAPK) sig naling pathway, H9C2 cells were cultured in vitro and divided into the control group which is without intervention, LPS groupwith 10 μg/mL LPS treatment, catechin with different concentrations (20, 40, 80, 160 nmol/L catechin treatment based on LPSgroup) and SB203580 group (10 μg/mL LPS+1 μmol/L SB203580 treatment), inhibitor group (10 μg/mL LPS+160 nmol/L catechin +1 μmol/L p38 MAPK pathway inhibitor SB203580 treatment) and activator group (10 μg/mL LPS+160 nmol/L catechin+10 μmol/L p38 MAPK pathway activator C16-PAF). Cell viability was measured with cell counting kit-8 (CCK-8) after 24 htreatment. The levels of interleukin-6 (IL-6) and interleukin-10 (IL-10) were detected by enzyme-linked immunosorbent assay(ELISA). Malondialdehyde (MDA) and superoxide dismutase (SOD) kits were used to detect the expression levels of oxidativestress factors MDA and SOD in cell supernatant. The apoptosis rate was determined by Hoechst 33258 staining. The expression levels of Caspase-3 and p38 MAPK pathway-related proteins were determined by Western blot. Compared with the controlgroup, the cell viability of LPS group significantly decreased (P<0.05). Compared with LPS group, the cell viability significantly increased after 160 nmol/L catechin supplementation (P<0.05). Finally, 160 nmol/L catechin group was selected as catechin group for follow-up experiment. In the follow-up experiment, compared with the control group, SOD and IL-10 contentsin LPS group significantly decreased (P<0.05), MDA and IL-6 contents, apoptosis number, Caspase-3 and P-P38 MAPK protein expression significantly increased (P<0.05)；compared with LPS group, catechin group and SB203580 group significantly reversed the changes of the above indexes (P<0.05)；compared with the catechin group, SB203580 in the inhibitor groupenhanced the changes in the above indicators, while C16-PAF in the activator group attenuated the effect of catechin on LPSinduced H9C2 cells (P<0.05). In this study, catechins can significantly inhibit the apoptosis, inflammation and oxidative stressinjury of H9C2 cells induced by LPS, and the mechanism of action may be related to the inhibition of p38 MAPK pathway signal transduction..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 259 (2024)
Effect and Mechanism of Azithromycin on Inflammatory Injury ofAlveolar Epithelial Cells Induced by Lipopolysaccharide
WEN Ling, LI Baoqi, ZHAO Yanmin, ZHENG Shuyang, and SU Ying
To explore the regulatory effects of azithromycin on lipopolysaccharide (LPS) -induced proliferation, apoptosisand Janus kinase 2 (JAK2)/signal transduction and transcription promoter 3 (STAT3) signaling pathways in human alveolarepithelial cells, in vitro culture of human alveolar epithelial cells A549, were divided iTo explore the regulatory effects of azithromycin on lipopolysaccharide (LPS) -induced proliferation, apoptosisand Janus kinase 2 (JAK2)/signal transduction and transcription promoter 3 (STAT3) signaling pathways in human alveolarepithelial cells, in vitro culture of human alveolar epithelial cells A549, were divided into blank group (no intervention), LPSgroup (10 μg/mL LPS treated for 24 h), low/medium/high concentration experimental group (10 μg/mL LPS+1, 2, 4 μg/mLazithromycin) and azithromycin group (10 μg/mL LPS+4 μg/mL azithromycin), inhibitor group (10 μg/mL LPS+50 μmol/LJAK2/STAT3 pathway inhibitor AG490), azithromycin+inhibitor group (10 μg/mL LPS+4 μg/mL azithromycin+50 μmol/LAG490) and azithromycin+activator group (10 μg/mL LPS+4 μg/mL azithromycin+0.5 μmol/L JAK2/STAT3 pathway activator Colivelin). After 24 hours of intervention, enzyme-linked immunosorption assay (ELISA), cell counting kit-8 (CCK-8),5-acetyney-2' deoxyuracil nucleoside (EdU), Hoechst 33258 staining and Western blotting (WB) were used for the expressionlevels of inflammatory factors interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), cell viability, proliferation rate, apoptosis rate, Caspase-3 and Cyclin D1 and the expression levels of JAK2/STAT3 signaling pathway related proteins. The results show that: compared with blank group, the expression levels of inflammatory cytokines IL-6, IL-8 and TNF-αin LPS group significantly increased, and cell viability significantly decreased. Compared with LPS group, the expression levelsof inflammatory cytokines IL-6, IL-8 and TNF-α in high-concentration experimental group significantly decreased, and cell viability significantly increased. In this study, 4 μg/mL azithromycin with significant difference from LPS group was selected asazithromycin group for follow-up experiments. Compared with blank group, the cell proliferation rate and Cyclin D1 protein expression levels in LPS group significantly decreased, while the apoptosis rate, Caspase-3, p-JAK2 and p-STAT3 protein expression levels significantly increased. Compared with LPS group, azithromycin group and inhibitor group significantly reversed thechanges of the above indexes. Compared with azithromycin group, cell proliferation rate and Cyclin D1 protein expression levels in azithromycin+inhibitor group further significantly increased, while apoptosis rate, IL-6, IL-8, TNF-α, Caspase-3, p-JAK2and p-STAT3 protein expression levels further significantly decreased. Azithromycin+activator group significantly reversed thechanges of the above indexes. Azithromycin can reduce LPS-induced inflammatory damage of A549 cells by inhibiting JAK2/STAT3 signaling pathway, promote cell proliferation and inhibit apoptosis, and provide evidence for exploring azithromycintreatment of LPS induced acute lung injury..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 267 (2024)
Protective Effects of Metformin Mediated NF-κB Signaling Pathway onCell Damage in Diabetic Nephropathy
LIU Danyang, WU Zhenyong, SUN Yaru, and CUI Yuxiu
In order to explore the protective effect of metformin mediated nuclear factor-κB (NF-κB) signaling pathway on thedamage of diabetic nephropathy cells, HGPC cells were cultured in vitro and treated with 10, 20, 30 and 40 mmol/L D-glucoseto select the optimal concentration of D-glucose for the construction of HGPC hyperIn order to explore the protective effect of metformin mediated nuclear factor-κB (NF-κB) signaling pathway on thedamage of diabetic nephropathy cells, HGPC cells were cultured in vitro and treated with 10, 20, 30 and 40 mmol/L D-glucoseto select the optimal concentration of D-glucose for the construction of HGPC hyperglycemic inflammatory injury model. Then20, 40, 80 and 160 mmol/L metformin were used for intervention, and the optimal concentration of metformin was screened.The cells were then divided into control group (Con group), high glucose group (HG group) and metformin group (Met group,30 mmol/L D-glucose+80 mmol/L metformin), inhibitor group (Y group, 30 mmol/L D-glucose+1 μmol/L NF-κB pathway inhibitor BAY 11-7082), metformin+inhibitor group (Met+Y group, 30 mmol/L D-glucose+80 mmol/L metformin+1 μmol/LBAY 11-7082) and metformin+agonist groups (Met+A group, 30 mmol/L D-glucose+80 mmol/L metformin+1 μmol/L NF-κBpathway agonist Prostratin), intervention was conducted for 24 h. Cell count kit 8 (CCK-8) was used to detect cell viability；theexpression levels of inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA)；cell invasion andmigration were determined by Transwell assay. Western blotting (WB) was used to detect the expression levels of E-cadherin,N-cadherin, vimentin, Fibronectin (FN), NF-κB p65 and p-NF-κB p65. The results show that: HGPC cells were treated with30 mmol/L D-glucose for 24 h to construct HGPC hyperglycemic inflammation model, and 80 mmol/L was the optimal concentration of metformin. Compared with Con group, the levels of inflammatory cytokines tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β), invasion ability, migration ability, the expression levels of N-cadherin, vimentin, FN and p-NF-κB p65protein in HG group significantly increased, while the expression level of E-cadherin protein decreased (P<0.05). Comparedwith HG group, the levels of inflammatory cytokines TNF-α, IL-1β, cell invasion ability, migration ability, the expression levels of N-cadherin, vimentin, FN and p-NF-κB p65 protein in Met group and Y group significantly decreased, and the expressionlevel of E-cadherin protein increased (P<0.05). Compared with the Met group, the above indexes changed more significantlyafter the addition of BAY 11-7082 (P<0.05), and the trend of Met+A group was opposite to that of Met+Y group (P<0.05).Metformin inhibits D-glucose-induced HGPC inflammatory response, invasion, migration and epithelial-mesenchymal transitionprocess by blocking the activation of NF-κB pathway, and protects podocyte injury induced by high glucose..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 275 (2024)
Effects of Oridonin on Migration and Invasion of HumanChondrosarcoma SW-1353 Cells
XU Chao, LI Zhen, MENG Di, and LI Jun
To investigate the effects of oridonin on migration and invasion of human chondrosarcoma cells. In vitro experiments, human chondrosarcoma SW-1353 cells were divided into control group being without intervention, 1.25, 2.50, 5.00μmol/L oridonin groups (1.25, 2.50, 5.00 μmol/L oridonin) and positive drug group (6 μmol/LTo investigate the effects of oridonin on migration and invasion of human chondrosarcoma cells. In vitro experiments, human chondrosarcoma SW-1353 cells were divided into control group being without intervention, 1.25, 2.50, 5.00μmol/L oridonin groups (1.25, 2.50, 5.00 μmol/L oridonin) and positive drug group (6 μmol/L adriamycin). Human chondrosarcoma SW-1353 cells were tested for viability, mobility, invasion ability and epithelial-mesenchymal transition (EMT) expression. The results show that: compared with the control group, SW-1353 cell viability in the 5.00 μmol/L oridonin group andpositive drug group significantly decreased (P<0.05). 1.25, 2.50, 5.00 μmol/L oridonin groups and positive drug obviously inhibted cell migration and invasion in a concentration-dependent manner (P<0.05). The expression of E-cadherin decreased theexpression of vimentin and N-cadherin and inhibited EMT process by increasing the expression of E-cadherin protein, and thehigher the concentration, the more significant the change was (P<0.05). Lamonidin can significantly inhibit the migration andinvasion of human chondrosarcoma SW-1353 cells, or through mediating EMT, which provides a theoretical basis for the biological study of human chondrosarcoma cells..
Acta Laser Biology Sinica
- Publication Date: Aug. 14, 2024
- Vol. 33, Issue 3, 284 (2024)
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