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RNA sequencing screening of differentially expressed genes after spinal cord injury

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Neural Regeneration Research 2019年第9期14卷 1583-1593页

作者： Yi Li Ying Chen Xiang Li Jian Wu Jing-Ying Pan ri-Xin Cai Ri-Yun Yang Xiao-Dong Wang School of Biology&Basic Medical Sciences Soochow UniversitySuzhouJiangsu ProvinceChina Department of Histology and Embryology Medical CollegeNantong UniversityNantongJiangsu ProvinceChina Jiangsu Key Laboratory of Neuroregeneration Co-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

In the search for a therapeutic schedule for spinal cord injury,it is necessary to understand key genes and their corresponding regulatory networks involved in the spinal cord injury process.However,ad hoc selection and analysis of one or two genes cannot fully reveal the complex molecular biological mechanisms of spinal cord injury.The emergence of second-generation sequencing technology(RNA sequencing)has provided a better method.In this study,RNA sequencing technology was used to analyze differentially expressed genes at different time points after spinal cord injury in rat models established by contusion of the eighth thoracic segment.The numbers of genes that changed significantly were 944,1362 and 1421 at 1,4 and 7 days after spinal cord injury respectively.After gene ontology analysis and temporal expression analysis of the differentially expressed genes,C5ar1,Socs3 and CCL6 genes were then selected and identified by real-time polymerase chain reaction and western blot assay.The mRNA expression trends of C5ar1,Socs3 and CCL6 genes were consistent with the RNA sequencing results.Further verification and analysis of C5ar1 indicate that the level of protein expression of C5ar1 was consistent with its nucleic acid level after spinal cord injury.C5ar1 was mainly expressed in neurons and astrocytes.Finally,the gene Itgb2,which may be related to C5ar1,was found by Chilibot database and literature search.Immunofluorescence histochemical results showed that the expression of Itgb2 was highly consistent with that of C5ar1.Itgb2 was expressed in astrocytes.RNA sequencing technology can screen differentially expressed genes at different time points after spinal cord injury.Through analysis and verification,genes strongly associated with spinal cord injury can be screened.This can provide experimental data for further determining the molecular mechanism of spinal cord injury,and also provide possible targets for the treatment of spinal cord injury.This study was approved ethically by the Laboratory Animal Ethics Commit

关键词： nerve regeneration spinal cord injury RNA sequencing C5ar1 Itgb2 Socs3 CCL6 astrocytes rats neural regeneration

Knockdown of polypyrimidine tract binding protein facilitates motor function recovery after spinal cord injury

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Neural Regeneration Research 2023年第2期18卷 396-403页

作者： Ri-Yun Yang Rui Chai Jing-Ying Pan Jing-Yin Bao Pan-Hui Xia Yan-Kai Wang Ying Chen Yi Li Jian Wu Gang Chen Department of Histology and Embryology Medical School of Nantong UniversityNantongJiangsu ProvinceChina Key Laboratory of Neuroregeneration of Jiangsu Province and the Ministry of Education Co-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina Center for Basic Medical Research Medical School of Nantong UniversityNantongJiangsu ProvinceChina Department of Anesthesiology Affiliated Hospital of Nantong UniversityNantongJiangsu ProvinceChina

After spinal cord injury(SCI),a fibroblast-and microglia-mediated fibrotic scar is formed in the lesion core,and a glial scar is formed around the fibrotic scar as a res ult of the activation and proliferation of astrocytes.Simultaneously,a large number of neuro ns are lost in the injured area.Regulating the dense glial scar and re plenishing neurons in the injured area are essential for SCI repair.Polypyrimidine tra ct binding protein(PTB),known as an RNA-binding protein,plays a key role in neurogenesis.Here,we utilized short hairpin RNAs(shRNAs)and antisense oligonucleotides(ASOs)to knock down PTB expression.We found that reactive spinal astrocytes from mice were directly reprogrammed into motoneuron-like cells by PTB downregulation in vitro.In a mouse model of compressioninduced SCI,adeno-associated viral shRNA-mediated PTB knockdown replenished motoneuron-like cells around the injured area.Basso Mouse Scale scores and forced swim,inclined plate,cold allodynia,and hot plate tests showed that PTB knockdown promoted motor function recovery in mice but did not improve sensory perception after SCI.Furthermore,ASO-mediated PTB knockdown improved motor function resto ration by not only replenishing motoneuron-like cells around the injured area but also by modestly reducing the density of the glial scar without disrupting its overall structure.Together,these findings suggest that PTB knockdown may be a promising therapeutic strategy to promote motor function recovery during spinal cord repair.

关键词： antisense oligonucleotides astrocytes glial scar motoneuron-like cells motor function neurogenesis neuron-like cells polypyrimidine tract binding protein short hairpin RNAs spinal cord repair

Facile and efficient synthesis of quinoline-4-carboxylic acids under microwave irradiation

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Chinese Chemical Letters 2010年第1期21卷 35-38页

作者： Zhu, Hui yang, ri Fang yun, Liu Hong Li, Jin Beijing Inst Pharmacol & Toxicol Dept Med Chem Beijing 100850 Peoples R China

A facile and efficient method for the preparation of 2-non-substituted quinoline-4-carboxylic acids is described via the Pfitzinger reaction of isatins with sodium pyruvate following consequent decarboxylation under m... 详细信息

关键词： Quinoline-4-carboxylic acids Pfitzinger reaction Microwave irradiation

Changes in human sperm motility and DNA fragmentation index after incubation at different temperatures following density gradient centrifugation and swim-up procedures

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Reproductive and Developmental Medicine 2022年第4期6卷 243-248页

作者： Yan-Nan yang Ling Wang Yu-Bing Liu yun-Jing Xue Chen-Chen Liu Fei Shi Xue Dai ri-Cheng Chian Center for Reproductive Medicine Shanghai 10th People’s Hospital of Tongji UniversityShanghai 200072China

Objective:The purpose of this study was to evaluate the sperm motility and DNA integrity at different temperatures to analyze whether the sperms are suitable on the second day for insemination of in vitro matured oocytes by intra-cytoplasmic sperm injection(ICSI)following density gradient centrifugation(DGC)and swim-up(SU)procedures.Methods:Semen samples were collected from 30 outpatients who visited the Center for Reproductive Medicine for semen analyses.Following sperm selection by DGC and SU procedures,the liquified semen samples were divided into three groups and incubated at 4,25,and 37°C,respectively.Following incubation for 24,48,and 72 hours,the sperm motility and sperm DNA fragmentation index(DFI)were analyzed.Results:Following the combination of DGC and SU procedures,the sperm motility(91.8%±8.6%vs.50.8%±13.1%)and DFI(5.1%±7.9%vs.13.0%±11.6%)were significantly improved(Prior incubation.However,sperm motility significantly increased(76.9%±10.4%)(Prison to that of the other 2 groups.However,the sperm DFI did not significantly increase when the sperm samples were incubated at 4(5.7%±5.9%)and 25°C(6.8%±5.6%)for 24 hours compared to that before incubation(5.1%±7.9%).Conclusions:These results indicate that the sperm quality,in terms of motility and DFI,can be efficiently improved by DGC in combination with SU.Following which,the sperm samples can be incubated at 25°C and be used on the second day for insemination of in vitro matured oocytes by ICSI.

关键词： Sperm motility Density gradient centrifugation Swim-up DNA fragmentation index matured oocytes

Deficiency of transmembrane AMPA receptor regulatory protein γ-8 leads to attention-deficit hyperactivity disorder-like behavior in mice

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Zoological Research 2022年第5期43卷 851-870页

作者： Wan-Jun Bai Xing-Guang Luo Bao-Hua Jin Kang-Sheng Zhu Wen-Yan Guo Xiao-Que Zhu Xia Qin Zu-Xiao yang Jiao-Jiao Zhao Si-Ruan Chen ri Wang Jie Hao Fei Wang yun Stone Shi De-Zhi Kong Wei Zhang Department of Pharmacology Institution of Chinese Integrative MedicineKey Laboratory of Neural and Vascular BiologyMinistry of EducationKey Laboratory of New Drug Pharmacology and ToxicologyHebei Medical UniversityResearch Unit of Digestive Tract Microecosystem Pharmacology and ToxicologyChinese Academy of Medical SciencesShijiazhuangHebei 050017China Department of Pharmacy Hebei General HospitalShijiazhuangHebei 050051China Beijing Huilongguan Hospital Peking University Huilongguan School of Clinical MedicineBeijing 100096China Fourth Hospital of Hebei Medical University ShijiazhuangHebei 050017China Department of Psychiatry Affiliated Nanjing Brain HospitalNanjing Medical UniversityNanjingJiangsu 210029China State Key Laboratory of Pharmaceutical Biotechnology and MOE Key Laboratory of Model Animal for Disease Study Model Animal Research CenterNanjing UniversityNanjingJiangsu 210061China

Attention-deficit hyperactivity disorder(ADHD) is a neurodevelopmental disorder prevalent in schoolage children. At present, however, its etiologies and risk factors are unknown. Transmembrane α-amino-3-hydroxy-5-methyl-4-isoxazolepropionicacid(AMPA) receptor regulatory protein γ-8(TARP γ-8,also known as calcium voltage-gated channel auxiliary subunit gamma 8(CACNG8)) is an auxiliary AMPA receptor(AMPAR) subunit. Here, we report an association between TARP γ-8 and ADHD,whereby adolescent TARP γ-8 knockout(KO) mice exhibitedADHD-likebehaviors,including hyperactivity, impulsivity, anxiety, impaired cognition,and memory deficits. Human single-nucleotide polymorphism(SNP) analysis also revealed strong associations between intronic alleles in CACNG8genes and ADHD susceptibility. In addition,synaptosomal proteomic analysis revealed dysfunction of the AMPA glutamate receptor complex in the hippocampi of TARP γ-8 KO mice.Proteomic analysis also revealed dysregulation of dopaminergic and glutamatergic transmissions in the prefrontal cortices of TARP γ-8 KO mice.Methylphenidate(MPH), which is commonly used to treat ADHD, significantly rescued the major behavioral deficits and abnormal synaptosomal proteins in TARP γ-8 KO mice. Notably, MPH significantly reversed the up-regulation of Grik2 and Slc6a3 in the prefrontal cortex. MPH also significantly improved synaptic AMPAR complex function by up-regulating other AMPAR auxiliary proteins in hippocampal synaptosomes. Taken together, our results suggest that TARP γ-8 is involved in the development of ADHD in humans.This study provides a useful alternative animal model with ADHD-like phenotypes related to TARP γ-8deficiency, which has great potential for the development of new therapies.

关键词： Attention-deficit hyperactivity disorder TARPγ-8 Methylphenidate Proteomics Phenome-wide association study

Downregulation of Na^＋/Ca^2＋ Exchanger Isoform 1 Protects Isolated Hearts by Sevoflurane Postconditioning but Not by Delayed Remote Ischemic Preconditioning in Rats

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Chinese Medical Journal 2017年第18期130卷 2226-2233页

作者： yang Yu Cheng-Hui Zhou yun-Tai Yao Li-Huan Li Department of Anesthesiology Fuwai Cardiovascular Hospital Beijing 100037 China

Background：Calcium regulatory proteins-L-type Ca^2＋ channels （LTCCs）,ryanodine receptor 2 （RyR2）,and Na^＋/Ca^2＋ exchanger isoform 1 （NCX1） have been recognized as important protective mechanisms during myocardial ischemia-reperfusion injury （I/ri）.Both sevofturane postconditioning （SevoPoC） and delayed remote ischemic preconditioning （DriPC） have been shown to protect the heart against I/ri.In this study,we aimed to compare the effects of SevoPoC and DriPC on the expression of the three calcium regulatory proteins in an isolated rat heart model.Methods：After 30-min balanced perfusion,isolated hearts from rats were subjected to 30-min ischemia followed by 60-min reperfusion.Totally 40 isolated hearts were randomly assigned to four groups （n =10/group）：time control group,I/ri group,SevoPoC group,and DriPC group.The effect of SevoPoC （3% v/v） and DriPC were observed.Myocardial infarct size （IS）,cardiac troponin I level,and heart function were measured.The protein and messenger RNA levels of LTCCs,RyR2,and NCX1 were determined.Results：Both SevoPoC and DriPC improved the recovery of myocardial function,and reduced cardiac troponin Ⅰ release after I/ri.The decrease in IS was more significant in the SevoPoC group than that in the DriPC group （16.50% ± 4.54% in the SevoPoC group [P =0.0006],and 22.34% ± 4.02% in the DriPC group [P =0.0007] vs.35.00% ± 5.24% in the I/ri group,respectively）.SevoPoC,but not DriPC significantly inhibited the activity of NCX 1 （0.59 ＋ 0.09 in the I/ri group vs.0.32 ± 0.16 in the SevoPoC group,P =0.006;vs.0.57 ± 0.14 in the DriPC group,P =0.072）.No statistical significant differences were observed in the expression of LTCCs and RyR2 between SevoPoC and DriPC.In addition,subsequent correlation analysis showed a significantly positive relationship between the cardiac troponin Ⅰ level and the protein expression ofNCX1 （r =0.505,P =0.023）.Conclusion：SevoPoC may be more effective in the cardioprotection than DriPC partly due to the deactivation of NCX1.

关键词： Calcium Regulatory Protein： Delayed Remote lschemic Preconditioning Ischemia-reperfusion Injury Na^＋/Ca^2＋ Exchanger Sevoflurane Postconditioning

A novel tube for sperm density gradient centrifugation procedure

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Reproductive and Developmental Medicine 2023年第4期7卷 224-229页

作者： yun-Jing Xue Wei Du Cun-Zhong Deng Yuan Xu Chen-Chen Liu Yan-Nan yang Ling Wang Yu-Zhen Lv Zheng Li ri-Cheng Chian Center for Reproductive Medicine Shanghai 1Oth People's Hospital of Tongji UniversityShanghai 200072China Center for Reproductive Medicine Women's and Children's Hospital of Jiaozuo CityJiaozuo 454001China Department of Andrology Center for Men's HealthShanghai General HospitalShanghai 200080China Department of ART Shanghai General HospitalShanghai200080China.

Objective:Density gradient centrifugation(DGC),swim up(SU),or a combination of DGC and SU is used to prepare sperm for infertility treatments using assisted reproductive technologies(ARTs).This study aimed to investigate the efficiency of a new device for detecting sperm DGC.Methods:Sperm samples were collected from 119 males who required semen analysis at three differentin vitro fertilization(IVF)centers(71 samples from Center A,28 samples from Center B,and 20 samples from Center C).Semen samples were divided into two groups for DGC:(1)a normal centrifugation tube and(2)a specially designed RC-Tube.The sperm concentration,motility,and DNA fragmentation index(DFI)were assessed using DGC.Sperm concentration and motility were assessed using a MAKLER counting chamber.Sperm DFI was measured and analyzed using flow cytometry using a DFI Kit.Results:There were no significant differences in sperm concentrations recovered following DGC between the two groups(31.1%±1.7%vs.28.8%±1.3%).Sperm motility was not significantly different following the DGC procedure in both groups(77.6%±1.6%.vs.75.8%±1.6%).The percentage of sperm DFI was also not significantly different following DGC between the two groups(10.4%±1.0%vs.10.5%±1.0%).Conclusions:The newly designed RC-Tube is efficient for sperm DGC and can be used to simplify sperm preparation for infertility treatment.

关键词： Sperm Centrifugation Motility DFI RC-Tube

Unique sandwich design of high-efficiency heat-conducting phosphor-in-glass film for high-quality laser-driven white lighting

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Journal of Advanced Ceramics 2022年第12期11卷 1889-1900页

作者： yang PENG Zikang YU Jiuzhou ZHAO Qing WANG Jiaxin LIU Bo SUN yun MOU Mingxiang CHEN School of Aerospace Engineering Huazhong University of Science and TechnologyWuhan 430074China School of Mechanical Science and Engineering Huazhong University of Science and TechnologyWuhan 430074China

Multi-color phosphor-in-glass(PiG)film has been considered as a promising color converter in high-quality laser lighting owing to its outstanding merits of phosphor versatility,tunable luminescence,and simple preparation.However,the opto-thermal properties of PiG film are severely affected by the photon reabsorption and backward scattering of phosphor structure and the heat conduction of substrate.Herein,a unique sandwich design of phosphor structure was introduced in the multi-color PiG film for high-quality laser lighting.By elaborately synthesizing the borosilicate glass with low glass transition temperature(T_(g)),similar expansion coefficient,and high refractive index(ri),the sandwiched PiGs were prepared by sintering(~600℃)broadband green and red phosphor glass films on the double sides of sapphire.The green and red PiG films were tightly coated on the sapphire with no delamination and maintained higher luminescence intensity than raw phosphors at high temperatures.By simultaneously coupling photon reabsorption and backward scattering,the sandwiched green PiG film-sapphire-red PiG film(G-S-R PiG)yields a high-quality white light with a high luminous efficacy of 163 lm/W and an excellent color rendering index(Cri)of 85.4 under a laser power of 2.4 W,which are the best comprehensive results yet reported.Benefiting from the ingenious sandwich design with heat-conducting sapphire and thin PiG films,the G-S-R PiG displays low working temperatures(<200℃)under high-power laser excitation.This work reveals the role of sandwiched phosphor structure in photon loss and heat dissipation,which provides a new strategy to design PiG films for high-quality laser lighting.

关键词： laser-driven lighting phosphor-in-glass(PiG)film sandwich structure color quality heat dissipation

Non-synchronous blade vibration analysis of a transonic fan

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Chinese Journal of Aeronautics 2023年第1期36卷 178-190页

作者： yun ZHENG Qingzhe GAO Hui yang School of Energy and Power Engineering Beihang UniversityBeijing 100191China Jiangxi Research Institute Beihang UniversityNanchang 330096China

This study numerically investigates the aeromechanic behavior of a transonic fan model with a flat tip-leading-edge on the NASA rotor 67 test case.Single-passage unsteady calculations at a near stall operating point of 82%design speed show that the dominant frequencies of mass flow were not the harmonics of the rotor rotational frequency.A full-annulus fluid–structure interaction analysis was subsequently carried out to examine the unsteady flows and their interactions with blade vibrations.The results show that the modal displacement of the backward traveling seventh nodal diameter of the second torsion mode grew exponentially,which reveals that the blade vibration was non-synchronous.The vibration pattern indicates that the aerodynamic mode was resonant with the structural vibration mode.Around the rotor tip,the circumferential vortical propagation induced by interactions among the main flow,tip leakage flow,and tip clearance vortex was the source of aerodynamic excitation.To clarify the mechanism of the non-synchronous vibration,the coupling between aerodynamic disturbance and structural response,i.e.,aliasing,was summarized.The frequency spectra of the fluctuating pressure show that an aerodynamic Back-ward Traveling Wave(BTW)was co-aliased to a structural BTW due to the propagation of the cir-cumferential vortex.The correlation between the frequency and free convective speed of the aerodynamic disturbance determined the directions of aliasing.

关键词： Aerodynamic disturbance Aliasing Blade vibration Circumferential vortical propagation Traveling wave