1.Construction of Saccharomyces cerevisiae Mutant Deficient in adh2 and ald6 Genes
Yan-Zun WANG ; Juan-Juan LEI ; Xian-Zhang JIANG ; Yuan-Yuan GAO ; Xin LI ; Can-Hua LAN ; You-Qiang CHEN ; Ru-Kai CHEN ; Jian-Zhong HUANG
Microbiology 1992;0(02):-
The purpose of this investigation is to improve ethanol production and decrease acetate formation in Saccharomyces cerevisiae strain YS2-?adh2.The strain YS2-?adh2 with deleted alcohol dehydrogenase Ⅱ(adh2) gene was isolated in our lab with higher ethanol production than that of the strain YS2.The ace-taldehyde dehydrogenase Ⅵ(ald6) gene encoded a cytosolic acetaldehyde dehydrogenase,a key enzyme of the pyruvate dehydrogenase(PDH) bypass,transfers acetaldehyde to acetate.To disrupt ald6 gene of the strain YS2-?adh2,ald6 gene targeting cassettes were synthesized by long flanking homology PCR(LFH-PCR) and then were transformed into YS2-?adh2 mutants by LiAc/SS Carrier DNA/PEG method.Positive transformants were selected with G418 and further confirmed by PCR.Once correctly integrated into the genome,the selective marker was rescued by transforming the plasmid pSH65 into the positive transformants and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.We named the ald6 gene knocked-out strain as YS2-?adh2-?ald6 which has a 12.5% higher ethanol production and a 18% lower acetate formation compared to the strain YS2.
2.Identication of pearl powder and conch powder from different origins by differential scanning calorimetry.
Jia CHEN ; Ming-hua LI ; Kun-zi YU ; Ya-juan DONG ; Nan-ping ZHANG ; Xiao-ru HU ; Feng WEI ; Shuang-cheng MA
China Journal of Chinese Materia Medica 2015;40(8):1459-1462
The paper is aimed to establish a methods for identication of pearl powder and conch powder from different origins. Hermetic aluminum pan was used to encapsulate samples. The optimal testing conditions were: heating rate 10 degrees C x min(-1), sample weight 3 mg and nitrogen gas flow rate 40 mL x min(-1). The enthalpy values of pearl powder and conch powder was obvious different. Identication of pearl powder and conch powder by DSC is a practical method for its accuracy, convenience and practificality.
Animal Shells
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chemistry
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Animals
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Calorimetry, Differential Scanning
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methods
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China
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Discriminant Analysis
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Pinctada
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chemistry
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classification
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Powders
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chemistry
3.Screening and validation of long non-coding RNAs in brain tissue of inflammation-induced preterm mice.
Ru-Juan CHEN ; Sha XI ; Fan WANG ; Mi XIAO ; Xiao-Jie LIN ; Li LIU
Chinese Journal of Contemporary Pediatrics 2016;18(5):435-439
OBJECTIVETo investigate the association between long non-coding RNAs (lncRNAs) and brain injury in inflammation-induced preterm mice, and to provide a reference for the prevention and treatment of brain injury.
METHODSAn intraperitoneal injection of lipopolysaccharide in pregnant mice was performed to establish a model of inflammation-induced preterm mice with brain injury (preterm group). The full-term mice delivered by normal pregnant mice were used as controls (full-term group). The lncRNA chip assay was used to screen out the lncRNAs associated with brain injury in preterm mice. Quantitative real-time PCR was used to validate the lncRNAs identified by the above method.
RESULTSThe preterm and full-term groups showed significant differences in the expression of 1 978 lncRNAs (P<0.05), consisting of 786 up-regulated lncRNAs and 1 192 down-regulated lncRNAs, and 29 lncRNAs were 1.5 or more times differentially expressed between the two groups. A further analysis was performed for the 10 most differentially expressed lncRNAs, and the results showed that these lncRNAs were involved in the biological processes including transcription, signal transduction, apoptosis, cell cycle, and inflammatory response, as well as G protein-coupled receptor signaling pathway and neuropeptide signaling pathway. Real-time PCR was performed to validate the expression of two lncRNAs in brain tissue in the preterm and full-term groups, and the results were consistent with those of the chip assay.
CONCLUSIONSThe expression profiles of lncRNAs in brain tissue change significantly in inflammation-induced preterm mice, and the G protein-coupled receptor signaling pathway may be involved in the pathogenesis of preterm brain injury.
Animals ; Brain ; metabolism ; Female ; Inflammation ; complications ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Long Noncoding ; analysis ; Receptors, G-Protein-Coupled ; physiology ; Signal Transduction ; physiology
4.Effects of Hand Robot-assisted Training Based on Motor Imagery on Upper Limb Function of Stroke Patients
Ying HOU ; Lin GAO ; Miao-Miao CHEN ; Ru-Jiao ZHANG ; Qing-Juan WANG
Chinese Journal of Rehabilitation Theory and Practice 2019;25(1):81-85
Objective To investigate the effect of hand robot-assisted training based on motor imagery on upper limb function of stroke patients.Methods From November, 2016 to May, 2018, 55 hemiplegic patients with upper limb dysfunction were randomly assigned to control group (n = 25) and experimental group (n = 30). The control group received routine hand motor training, while the experimental group received hand robot-assisted training, for four weeks. They were assessed with simplified Fugl-Meyer Assessment-Upper Extremity (FMA-UE), Wolf Motor Function Test (WMFT) and Modified Barthel Index (MBI) before treatment (t0), one week of treatment (t1), immediately after treatment (t2) and 2 months after treatment (t3).Results The score of WMFT improved in the experimental group at t1 (P < 0.05), with no significant difference between groups (Z =-0.901, P> 0.05). The scores of FMA, WMFT and MBI improved in both groups at t2 and t3 (P < 0.05), and the scores of FMA and MBI improved more in the experimental group than in the control group (Z>-2.073, t> 2.034, P < 0.05).Conclusion Hand robot-assisted training based on motor imagery can promote recovery of upper limb function in stroke patients more effective than routine hand function training.
5.Spatio-temporal process and the influencing factors on influenza A (H1N1) pandemic in Changsha
Hong XIAO ; Huai-Yu TIAN ; Jian ZHAO ; Xi-Xing ZHANG ; Pei-Juan ZHU ; Ru-Chun LIU ; Tian-Mu CHEN
Chinese Journal of Epidemiology 2011;32(6):587-592
Objective To analyze the spatio-temporal process on 2009 influenza A (HlNl) pandemic in Changsha and the influencing factors during the diffusion process. Methods Data were from the following 5 sources, influenza A (HlNl) pandemic gathered in 2009, Geographic Information System (GIS) of Changsha, the broad range of theorems and techniques of hot spot analysis, spatio-temporal process analysis and Spearman correlation analysis. Results Hot spot areas appeared to be more in the economically developed areas, such as cities and townships. The cluster of spatial-temporal distribution of influenza A (HlNl) pandemic was most likely appearing in Liuyang city (RR=22.70,P<0.01). The secondary cluster would include districts as Yuelu (RR=6A9,P< 0.01) , Yuhua (RR=81.63, P<0.01). Xingsha township appeared as the center in the Changsha county (RR=2.90, P<0.01) while townships as Yutangping (RR=19.31, P<0.01) , Chengjiao (RR=73.14,P<0.01) and Longtian appeared as the center in the west of Ningxiang county (RR= 14.43,P<0.01) and Wushan as the center in the Wangcheng county (RR= 13.84,P<0.01). As time went on, the epidemic moved towards the eastern and more developed regions. Regarding factor analysis, population, the amount of students, geographic relationship and business activities etc. appeared to be the key elements influencing the transmission of influenza A (H1N1) pandemic. At the beginning of the epidemic, population density served as the main factor (r=0.477, P<0.05) but during the initial and fast growing stages, it was replaced by the size of students to serve as the important indicator (r=0.831, P<0.01; r=0.518, P<0.01). However, during the peak of the epidemics, the business activities played an important role (r=-0.676, P<0.01). Conclusion Groups under high risk and districts with high incidence rates were shifting, along with the temporal process of influenza A(H1N1) pandemic, suggesting that the protection measures need to be adjusted, according to the significance of influencing factors at different stages.
6.Neuroprotective effect of Wendan Decoction on a mouse model of sleep disorders via IKKβ/NF-κB pathway
Li LI ; Ru LIU ; Jing HE ; Yun CHEN ; Juan GUO ; Ke JI ; Ling LIU
Chinese Traditional Patent Medicine 2024;46(3):803-809
AIM To investigate the effect of Wendan Decoction on nerve injury in a mouse model of sleep disorders and its mechanism.METHODS A mouse model of insomnia was established by the modified multiple platform sleep deprivation method.After successful modeling,the mice were randomly divided into the model group,the estazolam tablet group(0.15 mg/kg)and the low-dose and high-dose Wendan Decoction groups(12.5,50 g/kg),with 6 mice in each group,in contrast to the 6 mice of the control group.After 7 days of drug intervention,the mice had their changes of cerebral cortex,hippocampal CA1 area and hypothalamus observed by HE staining;their neuronal damage observed by Nissl staining;their levels of neurofilament light chain(NEFL),neuron-specific enolase(NSE),S100 calcium-binding protein B(S100B),tumor necrosis factor(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)in brain tissue and serum detected by ELISA;their cerebral expression of glial fibrillary acidic protein(GFAP)detected by immunohistochemical method;and their cerebral expressions of GFAP,phosphorylated IκB kinase β(p-IKKβ)and phosphorylated nuclear transcription factor-κB(p-NF-κB)detected by Western blot.RESULTS Compared with the model group,the high-dose Wendan Decoction group displayed increased number of neurons,complete and neatly arranged structure;decreased number of neurons with nuclear shrinkage and deformation;increased Nissl bodies,decreased levels of NEFL,NSE,S100B,TNF-α,IL-6 and IL-1β in serum and brain tissue(P<0.01);decreased cerebral expression of GFAP(P<0.01);and decreased phosphorylation levels of cerebral p-IKKβ and p-NF-κB(P<0.01).CONCLUSION Wendan Decoction can reduce the nerve damage and the expression of proinflammatory mediator in sleep disorders mice,and the mechanism may be related to the inhibited activation of IKKβ/NF-κB pathway.
8.Distribution of Inflammatory Cells and Expression of PSGL-1 in Infant Brainstem Tissue Related Fatal Brainstem Encephalitis.
Yan LIU ; Qiao-e ZHONG ; Jing-zai WANG ; Yong-zai WANG ; Jie GU ; Wen-juan SUN ; Hui-ru BAI ; Li-qin CHEN
Journal of Forensic Medicine 2015;31(5):347-351
OBJECTIVE:
To explore the distribution of inflammatory cells and positive expression of P-se- lectin glycoprotein ligand-1 (PSGL-1) in infant brainstem tissue from hand-foot-mouth disease related fatal brainstem encephalitis.
METHODS:
Twenty brainstem samples from infants suffered from brainstem en- cephalitis were collected as the experimental group. Ten brainstem samples from infants died of non- brain diseases and injuries were collected as the control group. The distribution of inflammatory cells and the expression of PSGL-1 in the two groups were examined by immunohistochemical method. The characteristics of the positive cells were observed.
RESULTS:
In brainstem tissue of the experimental group, there were sleeve infiltrations of inflammatory cells around the vessels and in the glial nodule. Microglia was the most and following was neutrophils around the vessels and in the glial nodule. There was a significant statistical difference among microglias, neutrophils and lymphocytes (P < 0.05). There was no sleeve infiltration in the control group. PSGL-1 protein was expressed widely in inflammatory cells in the experimental group, especially in the inflammatory cells around the vessels and in the glial nodule. But PSGL-1 positive staining could be observed significantly less in the control group comparing with the experimental group (P < 0.05).
CONCLUSION
Microglia is the main type of inflammatory cells involved in the progress of the fatal disease. Moreover, PSGL-1 could participate in the pathogenesis of hand-foot-mouth disease related fatal brainstem encephalitis.
Brain Stem/pathology*
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Encephalitis/pathology*
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Hand, Foot and Mouth Disease/pathology*
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Humans
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Infant
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Membrane Glycoproteins/metabolism*
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Microglia/pathology*
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Neutrophils/pathology*
9.Multivariate study on central line-associated bloodstream infection in patients undergoing hematopoietic stem cell transplantation
Ru-Hui HAN ; Mei-Juan JIN ; Xue-Feng QIAN ; Mei-Zhen QIAO ; Kai CHEN ; Chen WU
Chinese Journal of Infection Control 2019;18(2):127-131
Objective To investigate the incidence of central line-associated bloodstream infection (CLABSI) in patients with hematopoietic stem cell transplantation (HSCT), explore risk factors for the occurrence of CLABSI.Methods Basic information of patients with acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) who underwent HSCT in a hematology department from November 1, 2016 to October 31, 2017 was collected, incidences of original CLABSI (OCLABSI) and modified CLABSI (MCLABSI) were calculated, related risk factors were analyzed by multivariate Cox regression.Results A total of 218 patients with AML and MDS who underwent HSCT were enrolled, 19 of whom had OCLABSI and 10 had MCLABSI.Twenty-one strains of pathogens were isolated from 19 patients with OCLABSI, including 9 gram-positive bacteria, 11 gram-negative bacteria, 1 fungus;9 strains were multidrug-resistant organisms.The main risk factors for OCLABSI included the female (HR=0.088;95%CI:0.017-0.440;P=0.003), age (HR=1.560;95%CI:1.066-2.530;P=0.034), bone marrow cell transplantation only (HR=4.408;95%CI:1.860-22.593;P=0.043), ATG/CSA/MMF/MTXG for preventing graft-versus-host disease (GVHD) (HR=0.101;95%CI:0.015-0.686;P=0.019), and MTX for preventing GVHD (HR=0.097;95%CI:0.011-0.816;P=0.032).Conclusion Definition of MCLABSI can provide more accurate monitoring on deep central venous catheter-related bloodstream infection.Incidence of CLABSI in HSCT patients can be reduced by early detection of high-risk population according to high-risk factors, strict adherence to the prevention and control measures of bloodstream infection, and implementation of immune recombination after enhanced transplantation.
10.A novel biosynthetic hybrid scaffold seeded with olfactory ensheathing cells for treatment of spinal cord injuries.
Lei-min QIAN ; Zhi-jian ZHANG ; Ai-hua GONG ; Ru-juan QIN ; Xiang-lan SUN ; Xu-dong CAO ; Jin-bo LIU ; Ping JIANG ; Yong-chang CHEN
Chinese Medical Journal 2009;122(17):2032-2040
BACKGROUNDImplantation of tissue-engineered scaffolds is one of the most promising therapeutic strategies for inducing nerve regenerations following spinal cord injuries. In this paper, we report a novel bioengineered hybrid scaffold comprised of three major extracellular matrix (ECM) proteins.
METHODSECM-scaffolds (ECM-S) were prepared by gelling fibrinogen, fibronectin and laminin using fresh rat plasma. Olfactory ensheathing cells (OECs) were isolated from fresh rat olfactory mucosa, purified under differential adhesion, and assessed by immunofluorescent staining. OECs were seeded onto ECM-S and cultured. The effects of the scaffolds on the seeded cells were detected using the immunofluorescent staining, Western blotting, scanning electron microscopy and transmission electron microscopy.
RESULTSTissue-engineered ECM-S could be easily molded into mat-like or cylindrical shapes and gelled by addition of fresh plasma. Observations by electron microscopy show that the ECM-S forms a stable three-dimensional porous network. Studies on the effects of the ECM-S on the biological behaviors of OECs in vitro indicate that the scaffold can promote OEC adhesion, proliferation and process extensions. Additionally, OECs seeded on the scaffold maintained the expression of nerve growth factor, matrix metalloproteinase-3 and matrix metalloproteinase-9.
CONCLUSIONWe developed a biosynthetic hybrid gel which could be used as a scaffold for OEC transplantation; this gel can promote nerve regeneration following spinal cord injuries.
Animals ; Cells, Cultured ; Immunoblotting ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Olfactory Bulb ; cytology ; physiology ; transplantation ; Olfactory Mucosa ; cytology ; physiology ; transplantation ; Rats ; Spinal Cord Injuries ; therapy ; Tissue Engineering ; methods ; Tissue Scaffolds