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OBJECTIVE: To evaluate the extent of vascular endothelial dysfunction and preliminary identify serum protein biomarkers associated with obese individuals at risk for cardiovascular disease (CVD). METHODS: Fifteen obese volunteers with the phlegm-dampness constitution or balanced constitution were recruited for this study respectively. The clinical baseline data was collected, and the vascular endothelial function was evaluated using the EndoPATTM. Blood samples were collected for the serum proteome analysis. The differences in the serum protein expression levels between the two groups were detected and the protein interaction network analysis, correlation analysis, receiver operating characteristic (ROC) curve analysis, and random forest model investigation were conducted. RESULTS: There were no statistical differences found in the baseline data. For vascular endothelial function, the reactive hyperemia index (RHI) of the phlegm-dampness constitution obese group was significantly lower than that of the balanced constitution obese group (1.46 ± 0.30 vs 2.82 ± 0.78, P < 0.0001), indicating vascular endothelial dysfunction. There are 66 differentially expressed serum proteins between the two groups. apolipoprotein A2 (ApoA2), angiotensin-converting enzyme 2 (ACE-2), interleukin-33 (IL-33), and forkhead box P3 (FoxP3) showed significant differences and area under curve values of their ROC curves were greater than 0.7 and correlated significantly with RHI. CONCLUSION: Vascular endothelial dysfunction was present in the phlegm-dampness constitution obese group. Thus, alterations in the expression levels of key serum proteins, including ApoA2, ACE-2, IL-33, and FoxP3 could serve as potential biomarkers in the obese population at risk of CVD.
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Doenças Cardiovasculares , Medicina Tradicional Chinesa , Humanos , Proteoma/genética , Interleucina-33 , Obesidade , Biomarcadores , Proteínas Sanguíneas , Fatores de Transcrição ForkheadRESUMO
To promote the ideological and political construction of the doctor-patient communication course, the research group discussed the subject characteristics and proposed the goals, principles, elements, and paths of the ideological and political construction of the doctor-patient communication course combined with practical teaching and relevant policy documents. Besides, this paper put forward the top-level framework design for the implementation, curriculum assessment, and evaluation indicators of the ideological and political construction, and developed the Guidelines for Ideological and Political Teaching of the Doctor-patient Communication Course and related teaching evaluation indicators, with a view to providing reference evaluation standards for the ideological and political construction of the doctor-patient communication course in China.
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Objective:To analyze the level and latent categories of the training needs of the nursing staff of the social volunteer teams in Jiangsu Province, and to provide basis for the targeted training.Methods:From March to July in 2018, 224 elderly care teachers from 13 social volunteer teams of 100 Red Cross societies in Jiangsu Province were surveyed for the knowledge of the care for the aged and the needs of training by the self-designed questionnaires. A total of 207 valid questionnaires were collected, with an effective recovery rate of 92.4%. SPSS 21.0 and Latent GOLD 5.1 software were used for description analysis, hierarchical clustering and latent category analysis.Results:The number of answer of "very need" to knowledge and skills among the aged care teachers for the elderly, such as psychological care, elderly volunteer services and recreational activities for the elderly, was 196, 196 and 193, with a proportion of 94.7%, 94.7% and 93.2%, respectively. Cluster analysis showed that training needs could be divided into four aspects: life care knowledge and skills, health care knowledge and skills, psychological care knowledge and skills, and spiritual comfort knowledge and skills. The results of latent category analysis showed that the aged care teachers could be divided into three groups based on their different training needs: high demand group for overall knowledge and skills, partial demand group for overall knowledge and skills, and high demand group for medical care and psychological care knowledge and skills. The corresponding latent probability of the three groups were 80.2%, 13.0%, 6.8%, respectively.Conclusion:The training for the aged care of social volunteer teams should pay attention to the teaching of spiritual comfort knowledge and skills for the elderly. The training program should be oriented to the multi-level and individualized needs of the teachers, so as to meet the needs of the aged care teachers for their own development.
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Objective:To evaluate the retinal differentiation ability of human induced pluripotent stem cells (hiPSCs) from various somatic cell sources.Methods:The hiPSCs lines BC1- green fluorescent protein (GFP) and Gibco obtained by blood cell reprogramming and the hiPSCs line UE017 obtained by urine cell reprogramming were used to induce retinal differentiation.The morphogenesis and development of retina were recorded with an optical microscope, and the expression of specific molecular markers of various cell subclasses in the retina was detected by immunofluorescence, and the efficiency of retinal differentiation of different cell lines was analyzed and compared.Results:All three hiPSC lines derived from blood and urine cells were able to be induced into three-dimensional (3D) retinal organoids, including neuroretina and retinal pigment epithelial cells.Retinal organoids simulated the development process of retina in vivo and gradually differentiated into all cell subtypes of retina, including retinal ganglion cells, photoreceptor cells, amacrine cells, horizontal cells, bipolar cells, Müller cells, and even formed lamellar structures.However, in terms of the efficiency of acquiring retinal organoids, the hiPSCs derived from blood were more efficient than those derived from urine. Conclusions:hiPSCs from both blood and urine somatic cells can differentiate into 3D retinal organoids, including all subtypes of retinal cells.The differentiation efficiency among lines is different.
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Objective:To establish a fluorescent reporter human induced pluripotent stem cell line (hiPSCs) for monitoring the expression of visual system homeobox 2 ( VSX2). Methods:VSX2_small guide RNA (sgRNA) was inserted into vector PX459 to construct knockout plasmid, and the P2A-eGFP knock-in donor plasmid was conducted at the same time.The two plasmids were transfected into BC1-hiPSCs.Single cell clones were generated after treatment of puromycin.Correct insertion was confirmed by PCR and Sanger sequencing.The isogenicity of the parental and the reporter hiPSCs was confirmed by STR analysis and karyotyping.Pluripotency capacity of the reporter hiPSCs was analysed by reverse trascription PCR and immunofluorescence.Three-germ-layer formation experiment was carried out to analyse the multi-lineage differentiation ability of the reporter hiPSCs.The reporter hiPSCs were further differentiated to obtain three-dimension (3D) retinal organoids, and immunofluorescence was used to identify the co-localization of the enhanced green fluorescent protein (eGFP) and VSX2.Results:A VSX2 eGFP reporter hiPSC clone was successfully obtained by CRISPR/Cas9 technology, which was consistent with the parental hiPSCs (BC1-hiPSCs) in morphology, without any chromosomal aberrations or cell line cross-contamination.Reverse transcription PCR assay and immunofluorescence showed obvious positive expressions of iPSCs markers in BC1- VSX2 eGFP-iPSCs, including NANOG, OCT4, SOX2, DNMT3B and GDF3 mRNA as well as NANOG, OCT4, SSEA4 and TRA-1-60 protein.The α-fetoprotein (AFP), α-smooth muscle actin (α-SMA) and neuronal class Ⅲ β-tubulin (TUJ1) were expressed in endoderm, mesoderm and ectoderm, respeetively, derived from BC1- VSX2 eGFP-iPSCs, and eGFP and VSX2 were co-stained in the neural retinal layer of 3D retinal organoids derived from BC1- VSX2 eGFP-iPSCs by immunofluorescence. Conclusions:VSX2 fluorescent reporter hiPSCs is successfully generated, which can monitor the temporal and spatial expression changes of VSX2 protein in real time, providing a powerful tool for evaluation of retina development mechanism and cell therapy.
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Objective@#To observe the effect of monopolar radiofrequency(RF) and fractional CO2 laser on the filler hyaluronic acid (HA) injected subcutaneously in rats.@*Methods@#Five circular areas with a diameter of 2 cm at the dorsum of each rat were prepared for different treatment randomly: (1)HA injection group(HA). (2)HA injection+ fractional CO2 laser treatment group(HA+ CO2). (3)HA injection+ radiofrequency treatment group(HA+ RF). (4)0.9%NaCl+ fractional CO2 laser treatment group(NaCl+ CO2). (5)0.9%NaCl+ radiofrequency treatment group(NaCl+ RF). The volume of HA in group 1, 3, 4 were determined via MRI with reconstruction of Mimics Research X 17.0 before or 1, 3, 6, 12 months after photoelectric treatment. The rates of absorption of each time point and duration were calculated.@*Results@#The mass of HA could be even recognized 12 months after treatment macroscopicly. Inflamatory cell infiltration and fibring were observed around HA in HA, HA+ RF, HA+ CO2 group within 1 month. The fibrous capsule formed around HA, which was enhanced by RF and CO2 laser therapy since 3 months post therapy. There was no significant difference of volume of HA among the three groups before photoelectric treatment. There was significant difference of HA volume on 12 months post treatment, HA(311.90±52.59) μl, HA+ RF(404.80±64.29) μl, HA+ CO2(357.60±59.10) μl(P<0.01). The volume of HA received RF treatment degraded more slowly. The rate of HA absorption during 3-6 months post treatment was significantly lower than that of 6-12 months post treatment(P<0.05).@*Conclusions@#Fractional CO2 laser and radiofrequency do not increase the degradation of HA. On the contrary, they may prolong the duration of HA injected subcutaneously in rat.
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<p><b>OBJECTIVE</b>To observe the regional homogeneity (ReHo) of resting-state brain function in the healthy subjects of gastric distention treated with acupuncture at the back- and front- points of the stomach, Weishu (BL 21) and Zhongwan (CV 12) and the correlation with gastric motility so as to explore the mechanism on the central integration of the front- and back- points of the stomach.</p><p><b>METHODS</b>The crossover test design was adopted. Twenty-four healthy subjects were assigned to a Weishu group, a Zhongwan group and a combined-point group separately, 8 cases in each one in each of the three times. Totally, 24 subjects were included in each group. Under the water load condition, the subjects received acupuncture at Weishu (BL 21), Zhongwan (CV 12) and the combined Weishu (BL 21) and Zhongwan (CV 12). Before and after each acupuncture, the resting-state brain functional magnetic resonance imaging (fMRI) scan and electrogastrogram (EGG) test were applied. The ReHo value was calculated in the collected fMRI imaging data. The changes in ReHo values were analyzed and compared before and after acupuncture in each group, as well as among the groups. The gastric motility was analyzed before and after acupuncture. Additionally, the correlative analysis was conducted between the gastric motility and ReHo changes before and after acupuncture.</p><p><b>RESULTS</b>(1) After acupuncture, EGG amplitudes in the subjects of each group were lower remarkably as compared with those before acupuncture (all <0.01). The EGG frequencies were not different significantly as compared with those before acupuncture (all >0.05). The EGG amplitudes in the Weishu group and the Zhongwan group were higher than those in the combined-point group (both <0.05). (2) As compared with the conditions before acupuncture, acupuncture at the combined front- and the back- points as well as Weishu (BL 21) and Zhongwan (CV 12) separately all induced the changes in the brain ReHo. Acupuncture at the combined front- and the back- points significantly increased Reho values in the right inferior temporal gyrus, the left thalamus, the precuneus and the posterior cingulate gyrus (all <0.05) and remarkably reduced the ReHo values in the the middle temporal gyrus of the right temporal pole, sulcus calcarinus and precuneus (all <0.05). Compared with the single point groups, acupuncture at the combined front- and the back- points induced the increase of ReHo value in the posterior cingulate gyrus and the decrease of ReHo in the temporal pole (all <0.05). (3) The correlative analysis showed that the changes in the ReHo values in the posterior cingulate gyrus, the thalamus and the precuneus were positively correlated to the changes of the gastric motility amplitudes. The changes in the ReHo values in the temporal pole was negatively correlated to the changes of the gastric motility amplitudes.</p><p><b>CONCLUSION</b>Acupuncture at the combined back- and front- points of the stomach, as well as acupuncture at single Weishu (BL 21) and Zhongwan (CV 12) induce the ReHo changes in the different brain regions. Acupuncture at the combined back- and front- points of the stomach may induce the ReHo changes in some new brain regions as compared with the acupuncture at the single point. The thalamus, the posterior cingulate gyrus and the precuneus may be the the important integrated brain regions for acupuncture at the back- and the front- points in regulating the gastric motility. The effects of acupuncture at the back- and the front- points for the regulation of the gastric motility are closely related to the thalamus, the limbic system and the default network of the brain regions.</p>
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Humanos , Pontos de Acupuntura , Terapia por Acupuntura , Encéfalo , Diagnóstico por Imagem , Fisiologia , Estudos Cross-Over , Eletroencefalografia , Motilidade Gastrointestinal , Voluntários Saudáveis , Imageamento por Ressonância Magnética , EstômagoRESUMO
RT-PCR, Western blot, and immunofluorescence were used to examine secreted protein acidic and rich in cysteine(SPARC)mRNA and protein expression in adipose tissue of rats. The results showed that the expression levels of SPARC mRNA and protein in adipose tissue of insulin resistant rats were higher than those of the normal ones(all P<0.01). SPARC may play a role in the negative feedback regulation duning the development of insulin resistance.