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BACKGROUND:Compared with traditional two-dimensional culture,three-dimensional microtissue culture can show greater advantages.However,more favorable cultivation methods in three-dimensional culture still need to be further explored. OBJECTIVE:To evaluate the cell behavior of microtissue and its ability to promote cartilage formation under two three-dimensional culture methods. METHODS:Cartilage-derived microcarriers were prepared by chemical decellularization and tissue crushing.DNA quantification and nuclear staining were used to verify the success of decellularization,and histological staining was used to observe the matrix retention before and after decellularization.The microcarriers were characterized by scanning electron microscopy and CCK-8 assay.Cartilage-derived microtissues were constructed by combining cartilage-derived microcarriers with human adipose mesenchymal stem cells through three-dimensional static culture and three-dimensional dynamic culture methods.The cell viability and chondrogenic ability of the two groups of microtissues were detected by scanning electron microscopy,live and dead staining,and RT-qPCR. RESULTS AND CONCLUSION:(1)Cartilage-derived microcarriers were successfully prepared.Compared with before decellularization,the DNA content significantly decreased after decellularization(P<0.001).Scanning electron microscope observation showed that the surface of the microcarrier was surrounded by collagen,maintaining the characteristics of the natural extracellular matrix of cartilage cells.CCK-8 assay indicated that microcarriers had no cytotoxicity and could promote cell proliferation.(2)Scanning electron microscopy and live and dead staining results showed that compared with the three-dimensional static group,the three-dimensional dynamic group had a more extended morphology of microtissue cells,and extensive connections between cells and cells,between cells and matrix,and between matrix.(3)The results of RT-qPCR showed that the expressions of SOX9,proteoglycan,and type Ⅱ collagen in microtissues of both groups were increased at 7 or 14 days.The relative expression levels of each gene in the three-dimensional dynamic group were significantly higher than those in the three-dimensional static group at 14 days(P<0.05).At 21 days,the three-dimensional static group had significantly higher gene expression compared with the three-diomensional dynamic group(P<0.001).(4)The results showed that compared with three-dimensional static culture microtissue,three-dimensional dynamic culture microtissue could achieve higher expression of chondrogen-related genes in a shorter time,showing better cell viability and chondrogenic ability.
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OBJECTIVES@#To compare the clinical effect on Bell's facial palsy in the acute stage between the staging comprehensive treatment with acupuncture-moxibustion and western medication.@*METHODS@#Sixty patients with Bell's facial palsy in the acute stage were randomly divided into an observation group and a control group, with 30 cases in each one. The patients in the control group were administered orally with prednisone acetate tablets and methylcobalamin tablets until the 28th day of illness. In the observation group, the staging comprehensive treatment with acupuncture-moxibustion was adopted. On the affected side, Qianzheng (EX-HN 16), Yifeng (TE 17), Sibai (ST 2), Yangbai (GB 14), Jiache (ST 6), Dicang (ST 4) and Touwei (ST 8), etc. were stimulated. In the acute stage (Day 1 to 7 of illness), the routine acupuncture and the point-toward-point needle insertion were delivered, no any manipulation was exerted at acupoints, and the needles were retained for 30 min. In the subacute stage (Day 8 to 14 of illness), on the base of the treatment as the acute stage, the depth of needle insertion was adjusted at a part of acupoints and the even needling technique was operated by twisting needle. Besides, electroacupuncture (EA) was attached to Qianzheng (EX-HN 16) and Dicang (ST 4), with continuous wave of low intensity and high frequency, 100 Hz, for 20 min. In the recovery stage (Day 15 to 28 of illness), on the base of the treatment as the subacute stage, the heavy stimulation of acupuncture was given, in which, the sticking and lifting needle techniques were delivered after the needles were inserted from Sibai (ST 2) toward Dicang (ST 4), and from Dicang (ST 4) toward Jiache (ST 6), separately; warm needling was operated at Yifeng (TE 17), and EA changed to stimulate the acupoints with the intermittent wave of high intensity and low frequency, 2 Hz, for 30 min. Acupuncture-moxibustion was given once every other day until the end of the 28th day of illness. The level of House-Brackmann facial nerve function rating scale (H-B grade),the score of Sunnybrook facial nerve grading system (Sunnybrook), the score of facial disability index (FDI), the temperature difference in the infrared thermal imaging facial area and electromyogram (EMG) situation of the affected muscle group were observed before and after treatment in the two groups. Using musculoskeletal ultrasound,the facial nerve diameter was detected and the clinical effect was compared between the two groups.@*RESULTS@#After treatment, the level of H-B grade, Sunnybrook score, the scores of physical function and social life function in FDI were improved when compared with those before treatment in the patients of either group (P<0.01, P<0.05), and the results of these evaluations in the observation group were better than those of the control group (P<0.05). After treatment, the temperature difference of the frontal area, the eye area, the zygomatic area and the mouth corner was declined in comparison with that before treatment in the two groups (P<0.05), and the temperature difference in each area in the observation group was lower than that of the control group (P<0.05).The root mean square (RMS) of the frontal muscle group, the zygomatic muscle group and the orbicularis muscle group on the affected side increased in comparison with that before treatment in the two groups (P<0.01), and RMS of the observation group was higher than that of the control group (P<0.05) after treatment. Before treatment, the diameter of the facial nerve on the affected side was larger than that on the healthy side (P<0.01), and after treatment, the diameter on the affected side was reduced when compared with that before treatment in the two groups (P<0.01); the diameter of the facial nerve on the affected side in the observation group was smaller than that of the control group (P<0.05), while, the diameter on the affected side was larger when compared with the healthy side in the control group (P<0.05). The total effective rate of the observation group was 93.3% (28/30), higher than that of the control group (83.3% [25/30], P<0.05).@*CONCLUSIONS@#The staging comprehensive treatment with acupuncture-moxibustion is clearly effective on Bell's facial palsy in the acute stage, which affirms the effectiveness of acupuncture-moxibustion for the acute stage of Bell's facial palsy in comparison with conventional western medication.
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Humanos , Paralisia Facial/terapia , Moxibustão , Terapia por Acupuntura , Paralisia de Bell/terapia , FaceRESUMO
Objective:To study the in vitro construction of functional and self-renewing cartilage organoids based on cartilage acellular extracellular matrix (ECM) microcarriers.Methods:Fresh porcine articular cartilage was taken. The merely crushed cartilage particles were set as natural cartilage group and ECM microcarriers of appropriate particle size, which were prepared by the acellular method of combining physical centrifugation and chemical extraction, were set as microcarrier group. Cartilage organoids were constructed by loading human umbilical cord mesenchymal stem cells (hUCMSCs) and human chondrocytes (hCho) with a ratio of 3∶1 with microcarriers through a rotating bioreactor. The organoids with different induction times were divided into 0-, 7-, 14-, and 21-day induction groups. The cell residues of the microcarrier group and natural cartilage group were evaluated by 4′, 6-diaminidine 2-phenylindole (DAPI) fluorescence staining and DNA quantitative analysis. The retention of microcarrier components was observed by Safranin O and toluidine blue stainnings, and the collagen and glycosaminoglycan (GAGs) levels in the microcarrier group and the natural cartilage group were determined by colorimetric method and dimethyl-methylene blue (DMMB) method. The microcarriers were further characterized by scanning electron microscopy and energy dispersive spectroscopy. The hUCMSCs cultured with Dulbecco′s Modified Eagle′s Medium (DMEM) supplemented with fetal bovine serum (FBS) in a volume fraction of 10% was used as the control group and the hUCMSCs cultured with the microcarrier extract was used as the experimental group. Subgroups of hUCMSCs cultured at 3 time points: 1, 3 and 5 days were set up in the two groups separately. Cell Counting Kit 8 (CCK-8) was used to detect the biocompatibility of the two groups. The cellular activity of the organoids of the 0-, 7-, 14-, and 21-day induction groups was detected by live/dead staining and the self-renewal ability of the cartilage organoids of the 14-day induced group was identified by Ki67 fluorescence staining. The organoids of the 7-, 14-, and 21-day induction groups were detected by RT-PCR in terms of the expression levels of chondrogenesis-related marker aggrecan (ACAN), type II collagen (COL2A1), SRY-related high mobility group-box gene-9 (SOX9), cartilage hypertrophy-and mineralization-related marker type I collagen (COL1A1), Runt-related transcription factor-2 (RUNX2), and osteocalcin (OCN). Colorimetric and DMMB assays were performed to determine the ability of organoids in the 0-, 7-, 14-, and 21-day induction groups to secrete collagen and GAGs.Results:The results of DAPI fluorescent staining showed that the natural cartilage group had a large number of nuclei while the microcarrier group hardly had any nuclei. The DNA content of the microcarrier group was (7.8±1.8)ng/mg, which was significantly lower than that of the natural cartilage group [(526.7±14.7)ng/mg] ( P<0.01). Saffranin O and toluidine blue staining showed that the microcarrier was dark- and uniform-colored and it kept a lot of cartilage ECM components. The collagen and GAGs contents of the microcarrier group were (252.9±1.4)μg/mg and (173.4±0.8)μg/mg, which were significantly lower than those of the natural cartilage group [(311.9±2.2)μg/mg and (241.3±0.7)μg/mg] ( P<0.01). Scanning electron microscopy showed that the surface of the microcarriers had uneven and interleaved collagen fiber network. The results of energy spectrum analysis showed that elements C, O and N were evenly distributed in the microcarriers, indicating that the composition of the microcarrier was uniform. The microcarrier had good biocompatibility and there was no statistical significance in the results of CCK-8 test between the control group and the experimental group after 1 and 3 days of culture ( P>0.05). After 5 days of culture, the A value of the experimental group was 0.53±0.02, which was better than that of the control group (0.44±0.03) ( P<0.05). In the 0-, 7-, 14-, and 21-day induction groups, hUCMSCs and hCho were attached to the surface of the microcarriers, with good cellular activity, and the live/death rates were (70.6±1.1)%, (80.5±0.6)%, (94.5±0.9)%, and (90.8±0.5)% respectively ( P<0.01). There were a large number of Ki67 positive cells in cartilage organoids. RT-PCR showed that the expression levels of ACAN, COL2A1, SOX9, COL1A1, RUNX2 and OCN were 1.00±0.09, 1.00±0.24, 1.00±0.18, 1.00±0.03, 1.00±0.06 and 1.00±0.13 respectively in the 7-day induction group; 4.16±0.28, 5.09±1.25, 5.65±1.05, 0.47±0.01, 1.68±0.02 and 0.21±0.06 respectively in the 14-day induction group; 13.42±0.92, 3.07±0.21, 1.84±1.08, 2.72±0.17, 2.91±0.18 and 3.32±1.20 respectively in the 21-day induction group. Compared with the 7-day induction group, the expression levels of ACAN, COL2A1, SOX9 and RUNX2 in the 14-day group were increased ( P<0.05), but COL1A1 expression level was decreased ( P<0.05), with no significant difference in OCN expression level ( P>0.05). Compared with the 7-day induction group, the expression levels of ACAN, COL1A1 and RUNX2 in the 21-day induction group were significantly increased ( P<0.01), with no significant differences in the expression levels of COL2A1, SOX9 and OCN ( P>0.05). Compared with the 14-day induction group, the expression levels of ACAN, COL1A1, RUNX2 and OCN in the 21-day group were increased ( P<0.05 or 0.01), with no significant difference in the expression level of COL2A1 ( P>0.05), but the expression level of SOX9 was decreased ( P<0.05). The contents of collagen in 0-, 7-, 14-and 21-day induction groups were (219.15±0.48)μg/mg, (264.07±1.58)μg/mg, (270.83±0.84)μg/mg and (280.01±0.48)μg/mg respectively. The GAGs contents were (171.18±1.09)μg/mg, (184.06±1.37)μg/mg, (241.08±0.84)μg/mg and (201.14±0.17)μg/mg respectively. Compared with the 0-day induction group, the contents of collagen and GAGs in 7-, 14-, and 21-day induction groups were significantly increased ( P<0.01), among which the content of collagen was the lowest in 7-day induction group ( P<0.01) but the highest in the 21-day induced group ( P<0.01); the content of GAGs was the lowest in the 7-day induced group ( P<0.01) but the highest in the 14-day induction group ( P<0.01). Conclusions:The microcarriers prepared by combining physical and chemical methods are decellularized successfully, with more matrix retention, uniform composition and on cytotoxicity. By loading microcarriers with hUCMSCs and hCho, cartilage organoids are successfully constructed in vitro, which are characterized by good cell activity, self-renewal ability, strong expression of genes related to chondrogenesis and secretion of collagen and GAGs. The cartilage organoids constructed at 14 days of induction have the best chondrogenic activity.
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Neurexin-3α, discovered in 2016, is a new type of autoimmune encephalitis antibody. Anti-neurexin-3α antibody-associated encephalitis is generally associated with prodromal symptoms or mood changes, having main clinical manifestations as seizures, memory disorders, confusion or loss of consciousness, central ventilation insufficiency, abnormal behavior, and speech disorders. This paper reviews the relevant research progress at home and abroad about pathogenesis, diagnosis and differential diagnosis, treatment and prognosis of anti-neurexin-3α antibody-associated encephalitis, so as to expand the understanding of clinicians for this disease.
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@#Peri-implantitis and implant failure are common issues in the implant dentistry. The contaminated implant can be treated with mechanical, chemical and assisted methods for the formation of re-osseointegration. Re-osseointegration is achievable with valid bone regeneration strategies. The survival rate of implant into the site of previous failure is lower than that of the initial implant. For the hopeless and failed implant, it is necessary to remove the implant with corresponding technique.
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Leucine dehydrogenase (LDH) is the key rate-limiting enzyme in the production of L-2-aminobutyric acid (L-2-ABA). In this study, we modified the C-terminal Loop region of this enzyme to improve the specific enzyme activity and stability for efficient synthesis of L-2-ABA. Using molecular dynamics simulation of LDH, we analyzed the change of root mean square fluctuation (RMSF), rationally designed the Loop region with greatly fluctuated RMSF, and obtained a mutant EsLDHD2 with a specific enzyme activity 23.2% higher than that of the wild type. Since the rate of the threonine deaminase-catalyzed reaction converting L-threonine into 2-ketobutyrate was so fast, the multi-enzyme cascade catalysis system became unbalanced. Therefore, the LDH and the formate dehydrogenase were double copied in a new construct E. coli BL21/pACYCDuet-RM. Compared with E. coli BL21/pACYCDuet-RO, the molar conversion rate of L-2-ABA increased by 74.6%. The whole cell biotransformation conditions were optimized and the optimal pH, temperature and substrate concentration were 7.5, 35 °C and 80 g/L, respectively. Under these conditions, the molar conversion rate was higher than 99%. Finally, 80 g and 40 g L-threonine were consecutively fed into a 1 L reaction mixture under the optimal conversion conditions, producing 97.9 g L-2-ABA. Thus, this strategy provides a green and efficient synthesis of L-2-ABA, and has great industrial application potential.
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Aminobutiratos , Escherichia coli/genética , Leucina Desidrogenase/genética , Treonina DesidrataseRESUMO
Objective Toinvestigatetheeffectofpressureandnon-pressureonthedirectimagingoflowerlimbdeepveinswith CTvenography(CTV).Methods 100patients(50malesand50females,aged30-80yearsold,mean (63.5±13.5yearsold)with suspectedlowerextremityvenousdiseasesfrom September2015to October2018 wereretrospectivelyanalyzed.50patientswere scannedafterpressingtheankle(controlgroup),andtheother50werescannedwithoutpressingtheankle(experimentalgroup).Results Therewerenosignificantdifferencesbetweenthecontrolandexperimentalgroupsintheauxiliaryveinscore (t=-0.20,P=0.82), femoralveinscore(t=-0.1,P=0.91),andtotaliliacveinscore(t=-0.03,P=0.97).Conclusion CTV withoutpressingtheankle demonstratescomparableefficacytodirectimagingoflowerlimbdeepveins,withgoodimagequality,reducedcomplicationsandeasy toapply,sothatitshouldbewidelyusedinclinicalpractice.
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Objective To investigate the effect of modified radiographic position for peripherally inserted central catheter (PICC) on XGray imaging.Methods FiftyGone patients of standard anteriorGposterior radiography (control group)and 50 patients of radiography with the left side of the body tilted 10 to 30 degrees to the right (modified group)were collected retrospectively.The quality of imaging was analyzed between groups.Results The image quality was significantly improved in the modified group (χ2= 1 7.27,P<0.05).Conclusion When the bedside mobile DR is performed on PICC patients,raising the left side of the body and tilting to the right 10 to 30 degrees,known as modified radiographic position,can effectively increase the image quality and improve the identification of PICC,so as to accurately determine the depth of the PICC catheter.
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Objective Toinvestigatethechangesinbrainfunctionofheroindependentindividualsafternucleusaccumbens(NAC) ablativesurgery.Methods Twenty-fourheroindependentindividualsafterNACablativesurgery(NACgroup),27heroindependent individualsinshort-termabstinence(HDgroup)and32healthycontrol(HCgroup)wereincludedaresting-statefMRIstudy.Amplitudeoflow-frequencyfluctuations(ALFF)valuesofeachparticipantswerecalculated.DiffernecesinALFFvalueamongthethreegroupswere analyzed by One-Way ANOVA .Results Brain regions with statistically significant A L F F differences a m ong the three groups included left cerebellum,rightposteriorcentralgyrus,leftcaudatenucleus,rightmiddlefrontalgyrus,leftcuneusandbilateralinferiortemporal gyrusI.ntherightcentralposteriorgyrus,theALFFvalueofHDgroupwaslowerthanthatofNACandHCgroupI.ntheleftcerebellum,the ALFFvalueofHDgroupwashigherthanthatofNACandHCgrouprespectivelyI.nbilateralinferiortemporalgyrusandleftcaudatenucleus, ALFFvaluesofNACandHDgroupswerelowerthanHCgroup.ALFFvaluesofNACandHDgroupswerehigherthanHCgroup inrightmedialfrontalgyrusandleftcuneus.Conclusion InheroindependentindividualsafterNACablativesurgery,theALFFvaluesofleft cerebellumandrightposteriorcentralgyrustendtobenormal.NodifferenceinALFFvalueinregionsofdefaultmodenetworkbetweenNACand HDgroups.
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Objective To investigate the changes of brain white matter microstructures in patients with methamphetamine addiction and to analyze the correlation between the changes of brain white matter microstructures and mental symptoms. Methods Twenty-five methamphetamine addictions (MA) and twenty-five healthy controls (HC) were enrolled through community and treatment centres from 2015 to 2016. The data of diffusion tensor imaging (DTI) were obtained on 3.0 T MRI scanner. All the subjects′mental symptoms were assessed by symptom check list-90 (SCL-90). The fractional anisotropy (FA) was compared by the tract based spatial statistic (TBSS). For the comparison of the value of AD, the value of RD and clinical data between two groups, the normal distribution and homogeneity test of variance were carried out first. If the normal distribution and homogeneity of variance were satisfied, the two-sample t test was used, if not, the Mann-Whitney rank sum test was used. The correlation between FA, RD and AD values in white matter differential brain areas and methamphetamine dosage, time and psychosocial scale were analyzed by spearman test. Results SCL-90 scores in group MA were significantly higher than those in HC (P<0.05). Compared with HC group, FA value of right genu of corpus callosum in MA group decreased, and the value of RD value increased significantly (P<0.05). The AD value also decreased in MA group, but there was no significant difference between two groups(P>0.05). There was a positive correlation between FA value of white matter of differential brain area and hostility score in SCL-90 (r=0.450, P=0.024), and a negative correlation between RD value and hostility and paranoid scores (hostility:r=-0.590, P=0.002;paranoid:r=-0.438, P=0.028). Conclusion MA can damage the integrity of the synaptic fibers connecting the frontal lobe of the brain, which is related to multiple mental scores. It suggests that decreased white matter integrity may be one of the neurological mechanisms of mental disorders related to methamphetamine addiction.
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Objective To explore the difference in inhibitory control ability between DRD2 gene subtype of heroin addicts with fMRI.Methods Thirty-seven heroin-dependent patients were divided into DRD2 Taq IA+ group (A+,22)and matched DRD2 Taq IA-group (A-,15).Functional MRI was performed in all patients while they were executing an event-related go/nogo task at 3.0T MR scanner.The differences of brain activated images and behavioral data between the two groups were analyzed with SPM8 and SPSS1 6.0 software,respectively.Results There were no significant differences in reaction time,accuracy and false alarm rates between the two groups (P>0.05).Compared with A-group,weaker activation in the medial prefrontal cortex,dorsal anterior cingulate,middle cingulate,supplementary motor area,temporal lobe,fusiform gyrus,lingual gyrus,hippocampus and parahippocampal gyrus in response inhibition condition were demonstrated in A+ group (voxel number>228,t=2.11,Alphasim correction,P<0.05).There was no corrlation between the intensity within the activated brain regions and the usage of heroin,morphine urine test positive number(P>0.05).Conclusion DRD2 Taq IA+ group exist deactivation of the brain area on memory,inhibitory control,visual spatial attention. It may be the neural basis that contribute to easy addiction and relapse for A+ carriers.
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Objective To investigate the effects of different abstinent duration on brain functional network of heroin addicts at resting state.Methods Sixteen heroin addicts during protracted abstinence for 11-13 months (PA12 group) and twenty heroin addicts during protracted abstinence for 5-7 months (PA6 group) were recruited in the resting-state functional MRI study.Graph theoretical methods were applied to construct topological organization of whole brain network and nodes betweenness of the networks in all subjects,and the between-group differences were analyzed.The correlation of the node betweenness with the abstinence duration was conducted.Results There was no significant difference (P>0.05) in the small world characteristic (γ≈1,λ》1) between two groups.Compared with the PA6 group,the PA12 group demonstrated significantly decreased nodal betweenness in regions of left parahippocampal gyrus,left precentral gyrus,and significantly enhanced nodal betweenness in regions of the left cuneus,left temporal and right middle occipital gyrus (all P<0.05).Moreover,the betweenness of the left precentral gyrus (r =-0.52,P =0.001) and parahippocampal gyrus (r=-0.49,P=0.002) were negatively related with the abstinence duration,the betweenness of the right middle occipital was positively correlated with the abstinence duration (r=0.49,P=0.003).Conclusion The brain network small world topology of heroin addicts tend to be stable after 5-7 months of abstinence.Long-term abstinence may minimize the addicfs memory of drugs and potential drug seeking behavior,and recover addicts' visual spatial attention function.
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Objective To investigate the effect of constraint abstinence duration on regional homogeneity (ReHo) of heroin addicts in resting state.Methods Twenty heroin addicts abstinent for 6 months (PA6 group),sixteen heroin addicts abstinent for 11 months (PA11 group) and 30 demographically matched healthy controls (HC group) were enrolled.ReHo values were calculated and difference among three groups was analyzed.The relationship between ReHo values of differential brain regions and abstinent duration was performed with Pearson method.Results The results demonstrated statistically significant difference in ReHo values among three groups in the left parahippocampal gyrus,caudate,insula,cingulate gyrus,postcentral gyrus and right orbitofrontal cortex.Compared with the PA6 group,the PA11 group demonstrated lower ReHo value in left parahippocampal gyrus,caudate,insula,cingulate gyrus and postcentral gyrus,ReHos value were higher than PA6 and HC in right orbitofrontal cortex.Meanwhile,the ReHo values of left parahippocampal gyrus (r=0.53,P=0.000 8),caudate (r=0.54,P=0.000 6),insula (r=0.62,P<0.000 1),cingulate gyrus (r=0.47,P=0.003 5) and postcentral gyrus (r=0.35,P 0.034 8) were negatively correlated with abstinence duration and ReHo value of right orbitofrontal cortex (r=0.66,P<0.000 1) were positively correlated with abstinence duration.Conclusion During long-term abstinence,most of heroin addicts' damaged brain regions (mainly involved in craving,inhibitory control,learning and memory) tends to be normal,thus benefit reducing the risk of relapse,but motivation related brain region's (orbitofrontal cortex) neural activity increases abnormally,that may be one of the important reasons for relapse after abstinence.
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Psychological disorder has a high morbidity rate among patients with cardiovascular disease. Patients with cardiovascular disease associated with depression, especially post myocardial infarction and acute coronary syndromes associated with psychological disorder, have not only a poor prognosis, but also a high mortality . Therefore, comprehensive and perfect intervention is extremely important for patients with cardiovascular disease associated with psychological disorder. The intervention for such kind patients needs an integrated disease management. It needs to consolidate the treatment of body's disease to strengthen second-level prevention of heart disease, and pay attention to the intervention for psychological disorder as well.This paper aims to provide reference for reducing the negative effects of psychological illness in patients with cardiovascular disease during clinical treatment process, and make prospects about the significance of double heart medical model, through summarizing the interaction effects of psychological factors on patients with coronary heart disease, the significance ofdouble heartnursing mode and the specific nursing measures.
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Objective To explore the effect of methadone maintenace treatment (MMT)in functional connectivity (FC)of nucleus accumbens (Nacc)on heroin addicts,and to identify the potential neuromechanism of MMT performing on heroin craving.Methods Craving scores and resting-state fMRI (rs-fMRI)were performed in 37 heroin addicts under MMT and 26 matched heroin addicts (HA) without any treatment.The rs-fMRI data preprocessing was performed by data processing assistant for rs-fMRI (DPARSF)soft-ware based on Matlab 2009a.Bilateral Naccs were set as regions of interesting (ROIs)respectively,and then the mean time series and other voxels within whole brain were analyzed by the rs-fMRI data analysis toolkit (REST).Intra-and inter-group analysis was performed with a single sample t-test and two sample t-test respectively.The partial correlation between the intensity of FC in brain regions showed abnormal FC and the duration/doses of methadone consumption was further investigated by SPSS.Craving scores were tested with two sample t-test.Results Inter-group analysis showed the FC of the right Nacc with left dorsal medial/lateral prefrontal cortex and right dorsal anterior cingulate was significantly increased in the MMT group in comparisonwith HA group,how-ever,it was decreased with right medial orbitofrontal cortex.The FC between the left Nacc and left dorsal medial/lateral prefrontal cortex,right dorsal lateral prefrontal cortex,right dorsal anterior cingulate and left insular cortex was also significantly increased in the MMT group (P 26,t=2.91).There were no regions with induced FC.The craving scores of MMT were signifi-cantly lower than those of HA (t = - 2.03,P <0.05 ).There was no significant correlation between the abnormal FC and the duration/doses of methadone consumption in MMT group.Conclusion MMT may influence the function of the nucleus accumbens through cognitive control and motivation/drive circuits,thereby reduce drug craving of heroin addicts.
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Objective To explore the changes and effect of functional connectivity(FC) of reward network in chronic heroin addicts(HA) using resting-state fMRl(rs-fMRI). Methods rs-fMRI was performed on 26 chronic heroin addicts(HA group) and 25 matched normal volunteers(control group).The rs-fMRI data were proceeded by usingrs-fMRI(DPARSF) software on Matlab 2009a. Bilateral nucleus accumbens(Nacc) were set as ROIs. Then the mean time series of ROI were compared with other voxels within the brain by the rs-fMRI data analysis toolkit(REST). Intra-and inter-group analysis was performed with single sample and two sample t test respectively.The association of intensity of FC in brain regions and the duration/doses of heroin consumption was investigated. Results In HA group, positiveresting-state FC compared with the ROIs was found in frontal gyms, dorsal striatum, thalamus and pons, while temporal lobe, negative found in part of parietal cortex and occipital cortex, part of dorsal lateral prefrontal cortex and cerebellum. Within control group, positive FC compared with the ROIs was found in frontal gyms, dorsal striatum, thalamus, hippocampal gyrus and pons, occipital cortex, negative FC with ROIs found in part of dorsal lateral prefrontal cortex and cerebellum(P26). Compared with the control group,the FC between the right Nacc and the bilateral calcarinegyrus, left middle occipital gyrus(voxel numbe r=31,28,47;t=3.99, 3.74,3.74;and P0.05). Conclusion The abnormal interaction between Nacc and the regions involved in cognitive control and visual spatial attention may contribute to the heroin addiction.
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PurposeHeroin addiction is a chronic and recurrent functional brain disease, there are some functional changes in specific brain regions, but the network character remains unclear. The aim of this paper is to explore the network character of brain resting-state functional network in heroin addicts, to identify the potential neuromechanism of heroin addiction from the perspective of brain network.Materials and Methods Thirty heroin addicts (HA group) and twenty-nine healthy controls (control group) underwent resting-state functional MRI scanning using GE 3.0T MRI scanner. The brain functional networks were constructed based on graph theory, the small-world properties and node properties were calculated and compared between the two groups, the correlation between the total dosage of heroin and node degree was analyzed.Results Compared with control group, the small world characteristics of HA group was altered with statistically significant difference (P<0.05, corrected by false discovery rate); the node degrees in orbit frontal regions increased, while those in occipital brain regions decreased (P<0.05, corrected by false discovery rate). No correlation was found in HA group between node degree and the total dosage of heroin.Conclusion These results suggest that topology of functional brain networks were altered in heroin addicts which tends to random networks; increased motivational driving to the salience of drug and decreased visuospatial attention in heroin addicts may provide a strategy for identifying the neuromechanism of heroin addiction.
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Objective To explore the changes of cerebral local neuronal spontaneous activity between heroin dependent individuals and normal controls.Methods 3.0 T MRI were used for data acquisition,fMRI data were acquired during resting state from 20 male heroin dependent individuals(HD)and 1 5 matched normal controls(NC),and then the difference of amplitude of low frequency fluctuation (ALFF)between groups were analysised.Results Compared with NC,ALFF in rostral cingulated zone(bilateral anteri-or cingulate cortex ,medial prefrontal cortex )was significantly reduced in HD.Conclusion Dysfunction of resting state in RCZ were observed in HD,which may result in cognitive impairment and plays an important role in the transitions from ideas into behav-ior of relapse in addiction.
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Echinomycin is a small-molecule inhibitor of hypoxia-inducible factor-1 DNA-binding activity, which plays a crucial role in ovarian ovulation in mammalians. The present study was designed to test the hypothesis that hypoxia-inducible factor (HIF)-1alpha-mediated endothelin (ET)-2 expressions contributed to ovarian ovulation in response to human chorionic gonadotropin (hCG) during gonadotropin-induced superuvulation. By real-time RT-PCR analysis, ET-2 mRNA level was found to significantly decrease in the ovaries after echinomycin treatment, while HIF-1alpha mRNA and protein expression was not obviously changed. Further analysis also showed that these changes of ET-2 mRNA were consistent with HIF-1 activity in the ovaires, which is similar with HIF-1alpha and ET-2 expression in the granulosa cells with gonadotropin and echinomycin treatments. The results of HIF-1alpha and ET-2 expression in the granulosa cells transfected with cis-element oligodeoxynucleotide (dsODN) under gonadotropin treatment further indicated HIF-1alpha directly mediated the transcriptional activation of ET-2 during gonadotropin-induced superuvulation. Taken together, these results demonstrated that HIF-1alpha-mediated ET-2 transcriptional activation is one of the important mechanisms regulating gonadotropin-induced mammalian ovulatory precess in vivo.
Assuntos
Animais , Feminino , Humanos , Ratos , Células Cultivadas , Gonadotropina Coriônica/farmacologia , Equinomicina/farmacologia , Endotelina-2/genética , Gonadotropinas Equinas/farmacologia , Células da Granulosa/efeitos dos fármacos , Subunidade alfa do Fator 1 Induzível por Hipóxia/antagonistas & inibidores , Oligonucleotídeos/genética , Ovário/citologia , Ratos Sprague-Dawley , Superovulação/efeitos dos fármacos , Ativação TranscricionalRESUMO
Objective To clone the gene of arginine kinase (AK) from Periplaneta americana, produce its recombinant protein and investigate its allergenicity. Method The cDNA of AK was cloned using specific primers from the total RNA of P. americana. The cloned gene was inserted into pMD18-T vector and digested by BamHI and HindⅢ. The cDNA was sequenced and subcloned into pET-28a expression vector. The cloned AK cDNA gene was expressed in Escherichia coli BL21 (DE3) by IPTG induction. The recombinant AK (rAK) was purified by metal (Ni2+) chelating affinity chromatography. Its allergenicity was examined by both Western blotting and enzyme-linked immunosorbent assay (ELISA). Result The cloned cDNA ORF sequence (Accession no. EU429466) contained 1 068 bp and encoded 365 amino acids. Its sequence homology with the published one (Accession no. AY563004) was 99.9% at nucleotide level. The allergen rAK was highly expressed in E. coli BL21 (DE3) as a soluble protein mainly with the molecular weight of about Mr 45 000 under induction of IPTG and purified by 6-His-tag purification system. Both in the non-denaturalization and denaturalization conditions, the recombinant allergen was identified as its affinity to IgE antibodies from the cockroach-allergic patient sera by Western blotting and ELISA. Conclusion The recombinant cockroach arginine kinase has been obtained with proper allergenicity.