1.Role of glycogen synthase kinase-3β in trafficking of NR1-containing N-methyl-D-aspartate receptor and NR2B-containing N-methyl-D-aspartate receptor in spinal cord in rats with incisional pain and remifentanil-induced hyperalgesia
Yuan YUAN ; Jingyao WANG ; Fang YUAN ; Yonghao YU ; Guolin WANG
Chinese Journal of Anesthesiology 2012;32(7):787-790
Objective To investigate the role of glycogen synthase kinase-3β (GSK-3β) in the trafficking of NR1-containing N-methyl-D-aspartate receptor (NMDAR) and NR2B-containing NMDAR in the spinal cord in rats with incisional pain (IP) and remifentanil-induced hyperalgesia.Methods Twenty-four male Sprague-Dawley rats,aged 2-3 months,weighing 240-260 g,were randomly divided into 3 groups ( n =8 each):control group (group C),remifentanil group (group R) and GSK-3β inhibitor TDZD-8 group (group TDZD-8).A 1-cm longitudinal incision was made in the plantar surface of the left hindpaw in anesthetized rats.In group C,dimethyl sulfoxide (DMSO) 2 ml/kg was injected and then the normal saline (equal to the volume of remifentanil) was infused for 60 min via the caudal vein.In group R,DMSO 2 ml/kg was injected before IP was made and then remifentanil was infused at a rate of 1.2 μg· kg- 1 ·min- 1 for 60 min.In group TDZD-8,TDZD-8 2 ml/kg was injected before IP was made and then remifentanil was infused at a rate of 1.2 μg·kg-1 ·min-1 for 60 min.Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were measured 24 h before infusion of normal saline or remifentanil and at 2,6,24 and 48 h after intravenous injection.The rats were sacrificed after the last measurement of the pain threshold and L4-6 segment of the spinal cord was removed to determine the expression of NMDAR NR1 and NR2B subunits in cell membrane (s) and cytoplasm (i) in the spinal cord by Western blot analysis.The ratios of sNR1/iNR1 and sNR2B/iNR2B were calculated.Results Compared with group C,PWT was significantly decreased and PWL was significantly shortened,the expression of sNR1 and sNR2B was up-regulated,the expression of iNR1 and iNR2B was down-regulated,and the ratios of sNR1/iNR1 and sNR2B/iNR2B were significantly increased in groups R and TDZD-8 (P < 0.05).Compared with group R,PWT was significantly increased and PWL was significantly prolonged,the expression of sNR1 and sNR2B was down-regulated,the expression of iNR1 and iNR2B was up-regulated,and the ratios of sNR1/iNR1 and sNR2B/iNR2B were significantly decreased in group TDZD-8 ( P < 0.05).Conclusion GSK-3β is involved in the regulation of trafficking of NR1-containing NMDAR and NR2B-containing NMDAR from cytoplasm to cell membrane in the spinal cord in rats with IP and hyperalgesia induced by remifentanil.
3.Changes in total and surface N-methyl-D-aspartate receptor NR1, NR2A and NR2B subunits in spinal cord in a rat model of incisional pain-remifentanil-induced hyperalgesia
Yuan YUAN ; Jingyao WANG ; Fang YUAN ; Haiyun WANG ; Yonghao YU ; Guolin WANG
Chinese Journal of Anesthesiology 2011;31(9):1045-1048
Objective To investigate the changes in total and surface N-methyl-D-aspartate receptor (NMDAR) NR1,NR2A and NR2B subunits in spinal cord in a rat model of incisional pain(IP)-remifentanil-induced hyperalgesia.Methods Thirty-two male SD rate in which caudal vein catheter were successfully placed were randomly divided into 4 groups ( n =8 each):control group(group C),remifentanil group(group R),IP group (group I) and IP + remifentanil group (group R + I).In groups R and C remifentanil 1.2 μg· kg-1 · min-1 or the same volume of normal saline was infused intravenously for60 main.In groups R + I and I,a 1 cm longitudinal incision was made in the plantar surface of left hindpaw and remifentanil 1.2 μg· kg-1 ·min-1 or the same volume of normal saline was infused intravenously for 60 min.Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were measured at 24 h before and 2,6,24,48 h after remifentanil or normal saline administration.The animals were sacrificed after last pain threshold measurement.The L4-6 segment of the spinal cord was isolated for determination of the expression of the total and surface NMDAR NR1,NR2A and NR2B subunits in spinal cord by Western blot.The ratio of surface NR2B/NR2A was calculated.Results Compared with group C,PWT was significantly decreased,PWL shortened,the expression of the total and surface NMDAR NR1,NR2A and NR2B subunits up-regulated and the ratio of surface NR2B/NR2A increased in groups 1,R and R + I( P < 0.05).Compared with groups R and I,PWT was significantly decreased,PWL shortened,the expression of the total and surface NMDAR NR1,NR2A and NR2B subunits up-regulated and the ratio of surface NR2B/NR2A increased in group R + I( P < 0.05).There was no significant difference in the total and surface NMDAR NR2A subunits among the four groups( P > 0.05).Conclusion The up-regulation of total and surface NMDAR NR1 and NR2B subunits and the increased percentage of surface NR2B subunits in rats spinal cord may be involved in the development of incisional pain- remifentanil- induced hyperalgesia.
4.Assessment of the CT image quality for lung nodule using different field of view and matrix:a phantom study
Yuan FANG ; Xiangsheng XIAO ; Hong YU ; Jing LIU ; Liqing JIN
Journal of Practical Radiology 2016;32(10):1594-1597,1613
Objective To assess the CT image quality of the pure ground glass nodule(pGGN)in chest phantom by using different field of view(FOV)and matrix.Methods CT(Philips Brilliance 128 spiral CT)scans were performed on chest phantom containing 9 artificial pGGNs(diameter≥5 mm)in 3 different FOV (50 mm,150 mm,300 mm ),and were reconstructed in 2 different matrix(512×512, 1 024×1 024),standard kernel.Recorded the CT values and standard deviations (SD)of the nodules and surrounding regions in different FOV and matrix,and calculated the mean standard deviation(MSD),contrast noise ratio (CNR)and signal noise ratio (SNR),then compared the difference among them.Two radiologists assessed the image quality of the pGGNs in blind method respectively,then evaluated the coherence between them using Kappa testing.Results The coherence of 2 observers was substantial or almost perfect.No significant differences were found on MSD,CNR and SNR in different FOV (P value>0.05)when matrix was kept,whereas the visibility of the nodules improved with the FOV changing smaller.And there were significant differences all on MSD,CNR and SNR by using different matrix when FOV was kept.The MSD increased and the CNR,SNR decreased in high-resolution group.The visibility of the pGGNs was not improved obviously in high-resolution group.Conclusion The image quality of the pGGN(diameter≥5 mm)won’t be changed by using smaller FOV when matrix is kept ,but the visibility will be improved with the FOV changing smaller.When FOV is kept,the MSD increased and the CNR,SNR reduced in high-resolution group,but the visibility will not be improved obviously compare to the standard resolution group.
5.Construction of a micro-vascular network model in vitro by microfluidic technology for hemodynamic investigations
Yuan LI ; Beizhong LIU ; Fang GONG ; Chao YU ; Liyi HU
Military Medical Sciences 2014;(11):885-889
Objective The micro-vascular element plays a key role in the delivery of nutrients and the regulation of hemodynamic behavior, however, research is often hindered by ethical , economic and technological issues .Therefore, construction of a micro-vascular network in vitro will help to study the related pathological and physiological behavior in microvessels.Methods In this study, a micro-vascular element model with features of a micro-vascular network in vivo was designed based on the network structure of retinal arterioles .A micro-vascular network model in vitro, characterized by network asymmetry and the presence of both bifurcation-and side-branches , was developed by soft lithography technology . The developed microdevice allowed for the quantification of the cell -depletion layer ( CDL) thickness and hematocrit ( Ht) distribution within the microchannel networks .Results and Conclusion The study showed the potential of the developed in vitro model in revealing key hemodynamic features which have been detected for microvascular elements in vivo, including the relationships between CDL thickness , Ht and red blood cell distribution .The present study provides a new strategy and a technology for studying hemodynamics and microvascular system diseases in vitro.
6.The characteristics of cognitive impairment in patients with primary hypothyroidism
Yu MIAO ; Yuan YING ; Chi SONG ; Wang FANG
Chinese Journal of Behavioral Medicine and Brain Science 2013;(4):332-334
Objective To observe the characteristics of cognitive impairment in patients with primary hypothyroidism.Methods A total of 90 primary hypothyroidism patients untreated with thyroid hormone were selected.All the 90 patients were divided into subclinical hypothyroidism group,mild or moderate hypothyroidism group and serious hypothyroidism group,and 30 patients in each group.The other 30 healthy volunteers were selected as controls.Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were used for the testing their orientation,immediate memory,attention and calculation,delayed recalling,linguistic competence,visual space and execution,naming ability and abstracting power.One-way analysis of variance was performed to determine significant differences among four groups.Results The MMSE scores in subclinical hypothyroidism group,mild or moderate hypothyroidism group and serious hypothyroidism group((27.53 ± 2.16),(26.90±1.88) and (24.80 ± 2.10) respectively) were lower than those of the control (28.23 ± 1.33).The MoCA scores of the above hypothyroidism groups ((23.57 ± 3.33),(2 1.60 ± 2.81) and (20.53 ± 3.03) respectively) were also lower than that of the control (26.63 ± 2.31) (P < 0.05).Except for orientation and immediate memory,statistical significances of the other cognitive function were existed between hypothyroidism groups and the healthy controls (P< 0.05).With the increase in severity of hypothyroidism,the abnormality of attention,calculation,linguistic competence,visual space and executive ability,naming ability and abstracting power were appearing gradually in hypothyroidism groups (P < 0.05),and the scores were low(P< 0.05).Conclusion Defects of attention and calculation,delayed recalling,linguistic competence,visual space and execution,naming ability and abstracting power are existed in primary hypothyroidism patients.
7.Effect of eukaryotic expression plasmid containing methylenetetrahydrofolate reductase gene on transcriptional level of tumor-related genes in human gastric cancer cell line
Dan-Feng SUN ; Jing-Yuan FANG ; Yu-Rong WENG ;
Chinese Journal of Digestion 2001;0(10):-
Objective To analyze the effect of eukaryotic plasmids containing wild (sense) or anti- sense methylenetetrahydrofolate reductase (MTHFR) gene on cell viability and transcription level of tumor related genes in human gastric cancer cell line.Methods Human gastric cancer cell line MKN-45 was cultured.Recombinant plasmids containing wild MTHFR (W) or antisense MTHFR (A) gene, pCMV-W and pCMV-A,were constructed.Then pCMV-W,pCMV-A and pCMV blank plasmid were transfected into MKN45 cells respectively by using lipofect.Cell viability was analyzed by 3-(4,5-bime- thylthiazolyl-2)-2,5-diphenyhetrazolium dromide(MTT).The transcription levels of Dnmt 1,c-myc, p21~(WAF1) and hMLH1 genes were detected by real-time polymerase chain reaction(PCR).Results Cell vi- ability remarkably increased in those transfected with wild MTHFR (P<0.01),which was contrary to those transfected with antisense MTHFR(P<0.01).The expression of those tumor related genes mRNAs were all remarkably decreased in the MKN45-W cells in comparison with those in the MKN45-pCMV cells.No significant difference in the expressions of those tumor related genes mRNAs were found between the MKN45 cells transfected with pCMV-A and blank pCMV.Conclusion MTHFR influences cell viability and the expres- sion level of tumor related genes in human gastric cancer cell line MKN45.
8.Effect of Dexmedetomidine Combined Electrical Stimulation on Coanitive Function of Patients Receiving Extracerebral Intervention.
Jun YUAN ; Yu WU ; Ji-yong LI ; Xi CHEN ; Li ZHANG ; Yu-feng LIU ; Sheng-xiong TONG ; Fang-fang DENG
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(3):285-288
OBJECTIVETo explore the effect of dexmedetomidine combined electrical stimulation on cognitive function of neurosurgical diseases patients treated by extracerebral intervention.
METHODSTotally 122 patients with neurosurgical diseases who underwent selective intervention were randomly assigned to the observation group and the control group, 61 cases in each group. Patients in the control group recieved anesthesia by dexmedetomidine. Those in the observation group received electrical stimulation at Baihui (DU20), Yintang ( EX-HN3), and Neiguan (PC6) before dexmedetomidine anesthesia. The cognitive function of patients at preoperative day 1 and postoperative day 1 was respectively evaluated by Mini-Mental State Examinations (MMSE). Serum NSE, S-100β, IL-1β, IL-6, and TNF-α levels were detected in the two groups before intervention and immediately after intervention using ELISA.
RESULTSMMSE scores of two groups were significantly reduced at post-intervention day 1, as compared with one day before intervention. MMSE score of the observation group at post-intervention day 1 was (23.15 ± 1.87) points, significantly higher than that of the control group [ (19.34 ± 1.64) points , (P < 0.05)]. The postoperative cognitive dysfunction (POCD) incidence rate of the observation group was 16.4% (10/61), significantly lower than that of the control group [39.3% (24/61); P < 0.05]. Compared with before intervention, NSE and S-100β protein levels, IL-1β, IL-6 and α-TNF levels of the two groups increased (P < 0.05). Post-intervention NSE and S-100β protein levels, IL-1β, IL-6 and α-TNF levels were significantly lower in the observation group than in the control group (P < 0.05).
CONCLUSIONDexmedetomidine combied electrical stimulation could effectively prevent the occurrence of postoperative cognition, and reduce levels of NSA, S-100β, IL-1β, IL-6 and TNF-α.
Acupuncture Points ; Anesthesia ; methods ; Cognition ; Cognition Disorders ; prevention & control ; Dexmedetomidine ; therapeutic use ; Electric Stimulation Therapy ; Humans ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Neuropsychological Tests ; Neurosurgical Procedures ; Phosphopyruvate Hydratase ; blood ; Postoperative Complications ; Postoperative Period ; S100 Calcium Binding Protein beta Subunit ; blood ; Tumor Necrosis Factor-alpha ; blood
9.Evaluation of the preventive effect of DL0805-2 against monocrotaline induced rat pulmonary arterial hypertension
Di CHEN ; Tian-yi YUAN ; Yu-cai CHEN ; Hui-fang ZHANG ; Zi-ran NIU ; Lian-hua FANG ; Guan-hua DU
Acta Pharmaceutica Sinica 2021;56(1):208-216
In the treatment of hypertensive crisis, the novel Rho kinase inhibitor DL0805-2 can rapidly lower systematic blood pressure, reduce pulmonary artery pressure, and has a significant protective effect on lung injury. This experiment intends to evaluate the efficacy of DL0805-2 against pulmonary arterial hypertension (PAH) and preliminarily reveals its underlying mechanism. Animal welfare and experimental procedures are in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences. Sprague Dawley (SD) rats were randomly divided into DL0805-2 low, medium, and high dose groups (1, 3, and 10 mg·kg-1), bosentan positive control group, model group, and blank control group. The drug was administered daily on the 7th day after model establishment by monocrotaline injection. On the 25th day of the experiment, relevant indicators were examined to observe the therapeutic effect of DL0805-2 on pulmonary hypertension. DL0805-2 significantly relieved the abnormal changes in the physiological parameters related to PAH induced by monocrotaline, including reducing right ventricular systolic pressure, alleviating cardiac damage caused by pressure overload, and reducing the levels of endothelin-1 and inflammatory factors in lung tissues. DL0805-2 also attenuated pulmonary arteries remodeling. It was preliminarily discovered that DL0805-2 exerts preventive and therapeutic effect on PAH through Rho-kinase pathway. Our results suggested that DL0805-2 had good therapeutic effects on monocrotaline-induced PAH rat model. It intervened early in the disease process, effectively prevented the development of the disease, and reduced the mortality of the diseased animals. The mechanism is related to Rho-kinase pathway.
10.Central mechanisms of masticatory muscle pain induced by occlusal interference.
Yan-fang YU ; Zhi-yuan GU ; Kai-yuan FU
West China Journal of Stomatology 2007;25(6):588-590
OBJECTIVETo study c-fos and substance P expression in the central nervous system following mechanical and chemical nociceptive stimulation to the masseters in rats with occlusal interference.
METHODSOcclusal interference was made by bonding a 2 mm long dentin screw in the pulp cavity of the first maxillary molar in the left side. Seven days after occlusal interference, the rats in occlusal interference and mechanical stimulus group and mechanical stimulus control group were light anesthetized and nociceptive mechanical stimulus were applied to the ipsilateral masseter. Pain response was recorded and all the animals were killed 2 hours later. The rats in the other two groups were deep anesthetized and 100 microL 5% formalin was injected into the ipsilateral masseter, killed 2 hours later. The brainstem and cervical spinal cord were processed c-fos and substance P immunoreactivity and data were quantitatively analyzed.
RESULTSBoth mechanical and chemical stimulus to the ipsilateral masseter induced increasing neuronal c-fos expression in the trigeminal nucleus and in the cervical spinal dorsal horn in occlusal interference and mechanical stimulus group and occlusal interference and chemical stimulus group (P < 0.05). Following mechanical stimulation to the ipsilateral masseter, substance P expression in the trigeminal nucleus transition zone was increased in occlusal interference and mechanical stimulus group (P < 0.05).
CONCLUSIONThe central neuronal sensitization in the brainstem may play an important role in the masticatory muscle pain induced by occlusal interference.
Animals ; Masseter Muscle ; Masticatory Muscles ; Pain ; Proto-Oncogene Proteins c-fos ; Rats ; Rats, Sprague-Dawley