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1.
Pediatr Res ; 71(3): 235-40, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22258084

RESUMO

INTRODUCTION: Survivors of esophageal atresia and tracheo-esophageal fistula (EA-TEF) often suffer chronic respiratory tract disease. EA-TEF results from abnormal emergence of the trachea from the foregut. This study in a rat model tests the hypothesis that primary lung maldevelopment might be a downstream consequence of this defect. RESULTS: The lung was hypoplastic in rats with EA-TEF although the histological pattern was normal. Maturation and arteriolar wall thickness were unchanged, but mesenchymal control of airway branching was weakened. This branching was deficient from embryonal day (E13) on in adriamycin-treated explants. DISCUSSION: In conclusion, the lungs were hypoplastic in rats with experimental EA-TEF due to defective embryonal airway branching. However, arteriolar wall and respiratory epithelial patterns remained normal. These findings suggest that similarly defective lung development might contribute to chronic respiratory disease in EA-TEF patients. METHODS: Pregnant rats received either 1.75 mg/kg i.p. adriamycin or vehicle on E7, E8, and E9. Lungs were recovered at E15, E18, and E2. Lung weight/body weight ratio, total DNA and protein, radial alveolar count, arteriolar wall thickness, lung maturity, and mesenchymal control of airway branching were assessed. E13 lungs were cultured for 72 h and explant airway branching was measured daily. For comparisons, nonparametric tests (*P < 0.05) were used.


Assuntos
Atresia Esofágica/epidemiologia , Pulmão/anormalidades , Pulmão/patologia , Fístula Traqueoesofágica/epidemiologia , Animais , Doença Crônica , Comorbidade , Doxorrubicina/efeitos adversos , Atresia Esofágica/complicações , Feminino , Desenvolvimento Fetal , Pneumopatias/etiologia , Modelos Animais , Técnicas de Cultura de Órgãos , Gravidez , Ratos , Ratos Endogâmicos , Fístula Traqueoesofágica/complicações
2.
J Tradit Chin Med ; 42(6): 922-931, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36378050

RESUMO

OBJECTIVE: To investigate the efficacy of Yuzhizi seed extract (FAQSE) on inhibiting the proliferation of hepatocellular carcinoma (HCC) cells in vitro and to explore the anti-HCC action mechanism of FAQSE. METHODS: Human HCC HepG2 and Huh7 cells were used to investigate the anti-HCC effect of FAQSE. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) method was used to measure cell viability. Affymetrix microarray was adopted to detect the expression of transcriptome. The differentially expressed genes (DEGs) of each cell line were identified. For co-DEGs of both cell lines, the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were enriched using the Database for Annotation, Visualization and Integrated Discovery (DAVID), and the network analysis of protein-protein interaction (PPI) was mapped using the Retrieval of Interacting Genes/Proteins (STRING) and Cytoscape software. Some important genes in the PPI network of co-DEGs were selected to verify by quantitative real-time reverse transcription-polymerase chain reaction, Western blot and enzyme-linked immunosorbent assay. RESULTS: FAQSE decreased the viability of HepG2 and Huh7 cells. There were 211 co-upregulated and 86 co-downregualted genes in both cell lines after FAQSE treatment. The enriched GO terms of co-upregulated DEGs were primarily involved cell-cell adhesion, viral process, transcription initiation from RNA polymerase II promoter, positive regulation of transcription from RNA polymerase II promoter and actin cytoskeleton organization. The GO terms of co-downregulated DEGs were mainly enriched in the processes of SRP-dependent cotranslational protein targeting to membrane, viral transcription, nuclear-transcribed mRNA catabolic process, nonsense-mediated decay, translational initiation and rRNA processing. Main KEGG pathways of co-upregulated DEGs were endocytosis, glutathione metabolism, protein processing in endoplasmic reticulum, synaptic vesicle cycle and lysosome. The major KEGG pathways of co-downregulated DEGs were ribosome, biosynthesis of amino acids, arginine and proline metabolism, systemic lupus erythematosus and complement and coagulation cascades. The top 10 co-DEGs with high hub nodes in STRING analysis were ribosomal protein S27a, transferrin, ribosomal protein S20, ribosomal protein L9, protein phosphatase 2 regulatory subunit B alpha, transthyretin, thioredoxin reductase 1, ribosomal protein L3, ribophorin I and ribosomal protein L24. Alpha-fetoprotein (AFP) was also co-downregulated and contained in the PPI network. The mRNA and protein expression of most verified genes was consistent with the results of co-DEGs analysis. And the AFP level was significantly reduced after FAQSE treatment. CONCLUSIONS: A series of genes and pathways of HepG2 and Huh7 cells were changed after FAQSE treatment, which might be the targets of FAQSE against HCC and worthy of further study. AFP might be important one of them.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Transcriptoma , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , alfa-Fetoproteínas/genética , alfa-Fetoproteínas/metabolismo , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Redes Reguladoras de Genes , Biologia Computacional/métodos , Perfilação da Expressão Gênica/métodos , RNA Polimerase II/genética , RNA Polimerase II/metabolismo , Biomarcadores Tumorais/genética , Linhagem Celular , RNA Mensageiro , Extratos Vegetais/farmacologia , Regulação Neoplásica da Expressão Gênica
3.
Pediatr Surg Int ; 27(2): 175-80, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21069349

RESUMO

BACKGROUND/AIM: Infants and rats with congenital diaphragmatic hernia (CDH) have malformations of the heart and the great arteries caused by neural crest (NC) dysregulation during embryogenesis. Abnormally narrow jugular veins have been found in babies during cannulation for ECMO. However, the venous system has not been examined in depth so far. We hypothesized that abnormal patterning and/or size of the thoracic veins could occur in rats with CDH. This hypothesis was tested by microscopic magnetic resonance imaging (MMRI), a high-resolution tool able to detect subtle changes of vessels in small animals. MATERIAL/METHODS: Fetuses from pregnant rats fed either 100 mg i.g. nitrofen or vehicle on E9.5 were recovered near term. A 7 T MMRI system with a coronal multislice fast spin echo sequence allowed diagnosis of CDH (n = 19), and T2 SE high-resolution sequences made assessment of the pattern and width of cervico-thoracic veins possible. Values were corrected for body size by dividing them by the length of thoracic vertebrae T3-T5. The results in nitrofen and control (n = 11) groups were compared by non-parametric tests (*p < 0.05). RESULTS: Congenital diaphragmatic hernia fetuses were smaller than controls (4.5 ± 0.26 vs. 5.3 ± 0.2 g*). The widths (corrected for body size) of left external, both innominate, right superior vena cava and azygos veins were significantly smaller in CDH rats than in controls. CONCLUSIONS: The cervico-thoracic veins are normally patterned but abnormally narrow (except the internal jugulars) in rats with CDH. The same embryonic NC dysregulation that accounts for cardiovascular malformations could also explain these venous anomalies in CDH.


Assuntos
Veia Ázigos/embriologia , Veias Jugulares/embriologia , Imageamento por Ressonância Magnética/métodos , Microscopia/métodos , Prenhez , Animais , Veia Ázigos/anormalidades , Modelos Animais de Doenças , Feminino , Hérnia Diafragmática/patologia , Hérnias Diafragmáticas Congênitas , Veias Jugulares/anormalidades , Gravidez , Ratos , Ratos Sprague-Dawley
4.
Pediatr Surg Int ; 27(2): 165-73, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21069350

RESUMO

BACKGROUND/AIM: Esophageal dilatation, gastroesophageal reflux, and intestinal obstruction have been demonstrated in CDH survivors. Abnormal esophageal and intestinal innervations were recently found in rats and babies with this disease. Our aim was to further characterize these malformations in embryos and fetal rats exposed to nitrofen. METHODS: Pregnant rats received either 100 mg nitrofen or vehicle on E9.5. Fetuses were recovered at E15, E18, and E21. Sections of esophagus and small bowel were histochemically stained with acetylcholinesterase (AChE) and immunostained for PGP9.5. PGP9.5 gen protein were measured on E21 and PGP9.5 mRNA on E15, E18 and E21. Comparisons between groups were made with non-parametrics tests. RESULTS: Histochemistry and immunohistochemistry showed deficient innervation in all anatomical areas studied at E15, E18, and E21, and WB confirmed this decrease in E21 fetuses. PGP9.5 messenger was decreased in nitrofen-exposed animals on E18 (esophagus) or E15 (small bowel), and increased on E21 in the esophagus and E18 in small bowel. CONCLUSIONS: Development of the enteric nervous system of the esophagus, stomach, and small bowel is deficient in rat embryos and fetuses exposed to nitrofen. These anomalies could account in part for the long-term gastrointestinal morbidity observed in CDH survivors.


Assuntos
Sistema Nervoso Entérico/anormalidades , Prenhez , Animais , Western Blotting , Sistema Nervoso Entérico/embriologia , Sistema Nervoso Entérico/enzimologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Hérnia Diafragmática/embriologia , Hérnia Diafragmática/genética , Hérnias Diafragmáticas Congênitas , Imuno-Histoquímica , Gravidez , RNA/genética , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ubiquitina Tiolesterase/genética , Ubiquitina Tiolesterase/metabolismo
5.
Chinese Medical Ethics ; (6): 1200-1207, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1005581

RESUMO

【Objective:】 The co-word analysis method was used to analyze narrative medical literature studied in China in the past 10 years to explore the research status in this field and lay a foundation for further research. 【Methods:】 Using "narrative medicine" as the theme term, with the limited time interval of 2013—2022 to retrieve for relevant literature in CNKI, Wanfang, and VIP databases, and include literature that meets the research standards. Bicomb2 was used to perform keyword statistics and analysis on the downloaded literature. Ucinet6.0 and Netdraw were used to draw co-occurrence analysis graphs to visualize the co-occurrence relationship between high-frequency keywords. 【Results:】 A total of 565 papers were included and 31 high-frequency keywords were extracted. The co-occurrence relationship diagrams showed that the current research hotspots of narrative medicine in China include traditional Chinese medicine, nursing, general practice, evidence-based medicine, medical ethics, hospice care, chronic diseases, cancer and other popular disciplines and diseases. Most of them were about enhancing the empathy and humanistic caring ability of medical personnel and improving doctor-patient communication to improve the doctor-patient relationship, as well as doing well in medical humanities education and cultivating narrative ability through parallel medical records and reflective writing. The research on gerontology, improving the professional identity of medical staff, integrating literature and medicine, and the impact on patients were relatively few. 【Conclusion:】 At present, narrative medicine in China has achieved many achievements in fields such as nursing, traditional Chinese medicine, general practice medicine, chronic diseases, cancer, medical humanities education, and improving doctor-patient relationships. In the future, in-depth exploration can be conducted from the use of narrative medicine in multi-disciplinary and disease fields, trying different training methods, and synchronous cultivation of teachers and students, so as to build a more comprehensive narrative medicine system.

6.
Artigo em Inglês | WPRIM | ID: wpr-971088

RESUMO

OBJECTIVE@#To investigate the underlying molecular mechanisms by which silence information regulator (SIRT) 2 and glutaminase (GLS) in the amygdala regulate social behaviors in autistic rats.@*METHODS@#Rat models of autism were established by maternal sodium valproic acid (VPA) exposure in wild-type rats and SIRT2-knockout ( SIRT2 -/-) rats. Glutamate (Glu) content, brain weight, and expression levels of SIRT2, GLS proteins and apoptosis-associated proteins in rat amygdala at different developmental stages were examined, and the social behaviors of VPA rats were assessed by a three-chamber test. Then, lentiviral overexpression or interference vectors of GLS were injected into the amygdala of VPA rats. Brain weight, Glu content and expression level of GLS protein were measured, and the social behaviors assessed.@*RESULTS@#Brain weight, amygdala Glu content and the levels of SIRT2, GLS protein and pro-apoptotic protein caspase-3 in the amygdala were increased in VPA rats, while the level of anti-apoptotic protein Bcl-2 was decreased (all P<0.01). Compared with the wild-type rats, SIRT2 -/- rats displayed decreased expression of SIRT2 and GLS proteins in the amygdala, reduced Glu content, and improved social dysfunction (all P<0.01). Overexpression of GLS increased brain weight and Glu content, and aggravated social dysfunction in VPA rats (all P<0.01). Knockdown of GLS decreased brain weight and Glu content, and improved social dysfunction in VPA rats (all P<0.01).@*CONCLUSIONS@#The glutamate circulatory system in the amygdala of VPA induced autistic rats is abnormal. This is associated with the upregulation of SIRT2 expression and its induced increase of GLS production; knocking out SIRT2 gene or inhibiting the expression of GLS is helpful in maintaining the balanced glutamate cycle and in improving the social behavior disorder of rats.


Assuntos
Animais , Ratos , Tonsila do Cerebelo/metabolismo , Transtorno Autístico/metabolismo , Comportamento Animal , Modelos Animais de Doenças , Glutamatos/metabolismo , Glutaminase/metabolismo , Sirtuína 2/metabolismo , Comportamento Social
7.
Artigo em Chinês | WPRIM | ID: wpr-934022

RESUMO

Objective:To investigate the effects of long non-coding RNA (lncRNA) Gm13568 on the activation of A1 astrocytes and the progress of experimental autoimmune encephalomyelitis (EAE) in mice.Methods:A recombinant lentiviral vector (LV-Inhibit-Gm13568) carrying astrocyte-specific promoter of glial fibrillary acidic protein (GFAP) was established to inhibit the function of endogenous Gm13568. A control vector (LV-ctrl) was established as well. The recombinant vectors were packaged. C57BL/6 mice were injected with 1×10 7 transforming units of viral suspension via the tail vein and 7 d after the injection, myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55) was used to establish the mouse model of EAE. Four groups, PBS group, EAE group, LV-ctrl+ EAE group and LV-Inhibit-Gm13568+ EAE group, were included in this study. Clinical signs of the mice were monitored daily in a double-blinded manner. The mice were sacrificed 23 d after the EAE model was established and the spinal cord tissues were collected. The expression of Serping 1, C3, Srgn and H2-T23 at mRNA level was detected by real-time PCR. Changes in the expression of IL-6, TNF-α, macrophage chemotactic protein-1 (MCP-1) and interferon-inducible protein-10 (IP-10) were measured. Western blot was used to investigate the expression of GFAP and Notch1 in spinal cord tissues and the phosphorylation of signal transduction and transcription activator 3 (STAT3). The expression of Notch1 intracellular domain (NICD) and GFAP in spinal cord tissues was detected by immunofluorescence. Furthermore, the infiltration of inflammatory cells and the demyelination of spinal cord were observed using HE and Luxol fast blue (LFB) staining methods. Results:Compared with PBS group, A1 astrocytes were activated and Notch1 expression was significantly up-regulated in EAE group and LV-ctrl+ EAE group. The clinical score of mice in LV-Inhibit-Gm13568+ EAE group was decreased from an average score of 3.5 to less than 1 on 23 d after antigen induction and the clinical symptoms were alleviated as compared with the mice in LV-ctrl+ EAE group. Meanwhile, the activation of A1 astrocytes was down-regulated, and the production of inflammatory cytokines and chemokines was also reduced. The expression of Notch1, GFAP and NICD at protein level and the phosphorylation of STAT3 were significantly reduced. Moreover, the infiltration of inflammatory cells and demyelination of spinal cord tissues were alleviated significantly.Conclusions:LncRNA Gm13568 might regulate the activation of A1 astrocytes via the Notch1/STAT3 pathway, thus affecting the production of inflammatory cytokines and chemokines and participating in the process of EAE.

8.
Frontiers of Medicine ; (4): 276-284, 2022.
Artigo em Inglês | WPRIM | ID: wpr-929185

RESUMO

We aimed to evaluate the effectiveness and safety of single-course initial regimens in patients with low-risk gestational trophoblastic neoplasia (GTN). In this trial (NCT01823315), 276 patients were analyzed. Patients were allocated to three initiated regimens: single-course methotrexate (MTX), single-course MTX + dactinomycin (ACTD), and multi-course MTX (control arm). The primary endpoint was the complete remission (CR) rate by initial drug(s). The primary CR rate was 64.4% with multi-course MTX in the control arm. For the single-course MTX arm, the CR rate was 35.8% by one course; it increased to 59.3% after subsequent multi-course MTX, with non-inferiority to the control (difference -5.1%,95% confidence interval (CI) -19.4% to 9.2%, P = 0.014). After further treatment with multi-course ACTD, the CR rate (93.3%) was similar to that of the control (95.2%, P = 0.577). For the single-course MTX + ACTD arm, the CR rate was 46.7% by one course, which increased to 89.1% after subsequent multi-course, with non-inferiority (difference 24.7%, 95% CI 12.8%-36.6%, P < 0.001) to the control. It was similar to the CR rate by MTX and further ACTD in the control arm (89.1% vs. 95.2%, P =0.135). Four patients experienced recurrence, with no death, during the 2-year follow-up. We demonstrated that chemotherapy initiation with single-course MTX may be an alternative regimen for patients with low-risk GTN.


Assuntos
Feminino , Humanos , Gravidez , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Dactinomicina/efeitos adversos , Doença Trofoblástica Gestacional/tratamento farmacológico , Metotrexato/uso terapêutico , Estudos Retrospectivos
9.
Journal of Chinese Physician ; (12): 830-833, 2020.
Artigo em Chinês | WPRIM | ID: wpr-867317

RESUMO

Objective:To explore the association between vitamin A, E and recurrent respiratory tract infections (RRTIs) in children.Methods:1 853 children with RRTIs and 2 695 health children were separately selected as cases and controls. Propensity score matching (PSM) analysis were conducted to reducing confounding bias between groups. Blood samples were collected to measure serum levels of vitamins A and E by high performance liquid chromatographie (HPLC). Paired logistic regression was used to determine odds ratios ( OR) and 95% confidence intervals ( CI) to evaluate the association of RRTIs with the serum levels of vitamin A and E. Results:After propensity score estimation and 1∶1 matching, the study cohort included 1 507 cases with RRTIs and 1 507 health children as controls. The main characteristics between the two groups were all not significantly different after matching ( P>0.05). The average levels of vitamin A in two groups were different ( P<0.05). Paired logistic regression analysis showed that greater serum levels of vitamin A had a lower risk of RRTIs ( OR=0.46, 95% CI: 0.22-0.98), similarly, deficiency of vitamin A would also increase the risk of RRTIs ( OR=1.18, 95% CI: 1.00-1.38). The statistical difference was not found between VitE and RRTIs ( P>0.05). Conclusions:The serum levels of vitamin E didn′t have a statistically significant association with RRTIs. However, we observed an obvious association between vitamin A and RRTIs. Hence vitamin A clinical monitoring and supplementation are vital for preventing and treating RRTIs.

10.
Artigo em Chinês | WPRIM | ID: wpr-865835

RESUMO

Experimental Traditional Chinese Medicine is an emerging discipline that plays an important role in cultivating innovative talents of traditional Chinese medicine (TCM). In recent years, with the rapid development of TCM and the new requirements of positioning, and also combined with the college students' cognitive characteristics, we have revised and republished the "Experimental TCM" (Third Edition) textbook, which focuses on introducing knowledge by adopting relevant scientific research cases. This test book was used in the teaching of undergraduates of batch 2013 in the eight-year program in Shanghai University of Traditional Chinese Medicine. After-class questionnaires showed that this teaching mode, guided by scientific research case, is not only helpful for students to develop their quality and ability of adopting modern experimental methods initially in the study and development of TCM, but also able to spread the great achievements of TCM researches. The teaching mode is also conducive to enhancing students' sense of responsibility for the modernization of TCM. Therefore, it is suggested that the course of Experimental TCM should be promoted in the colleges and universities of TCM.

11.
Artigo em Chinês | WPRIM | ID: wpr-865797

RESUMO

Objective:To evaluate the teaching effect of moral education materials implied in scientific research cases in the teaching of "Experimental Traditional Chinese Medicine (TCM)".Methods:The moral education materials implied in scientific research cases of "Experimental TCM" were extracted and skillfully integrated into the teaching of professional knowledge. A questionnaire survey was conducted with questions as "whether it is good to demonstrate the potential humanistic spirit by case teaching, whether this helps improve your interest in science and whether this teaching form affects your study, life and work attitude in the future" "Ten specific items from scientists' moving deeds that touch students and their recognitions" to assess the teaching effect.Results:95.8 percent of students affirmed this teaching form and thought it helped improve their interest in scientific exploration. 87.5 percent of students considered the humanistic spirit would affect their study, life and work in the future. 77.1-89.6 percent of students held a positive attitude to the ten items derived from the scientists' moving stories. Among these items, the percentages of the two items, "the spirit of being able to endure loneliness, work hard to make contributions selflessly without seeking for rewards or reputations" and "having respect forpeople and their scientific research achievements with courage to challenge the authorities of scientific research" are the lowest and highest, respectively.Conclusion:It is good to apply the moral education materials implied in the scientific research cases in the teaching of "Experimental TCM", which basically achieves the teaching goal, but there is still room for improvement.

12.
China Pharmacy ; (12): 2473-2480, 2020.
Artigo em Chinês | WPRIM | ID: wpr-829353

RESUMO

OBJECTIVE:To identify t he chemical constituents of Fengliaoxing fengshi dieda wine. METHODS :An ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry (UPLC-Q-Exactive- MS)technique was used for identifying chemical constituents of Fengliaoxing fengshi dieda wine. The determination was performed on Thermo Accucore aQ RP 18 column with mobile phase consisted of 0.1% formic acid-methanol (gradient elution )at the flow rate of 0.3 mL/min. The column temperature was 30 ℃,and the sample size was 2 μL. HESI source was adopted,ion mode was positive and negative ion mode ,nitrogen as sheath gas and auxiliary gas. The positive ion mode had spray voltage of 3.5 kV, capillary heating temperature of 350 ℃,sheath gas pressure of 35 psi,auxiliary gas pressure of 15 arb,and ion source heating temperature of 320 ℃. The negative ion mode had spray voltage of 3.2 kV,capillary heating temperature of 350 ℃, sheath gas pressure of 35 psi,auxiliary gas pressure of 15 arb,and ion source heating temperature of 300 ℃. The mass axis was calibrated by external standard method (mass error less than 5 ppm). The scanning range of the first mass spectrometry was m/z 80.0-1 200.0 (the resolution was 70 000),the scanning range of secondary mass spectrometry was m/z 80.0-1 200.0(the resolution was 17 500),and the collision voltage was 20,40,60 eV. Retrieved from CNKI ,VIP,PubMed and other database ,the chemical constituents information of each Chinese traditional medicine in Fengliaoxing fengshi dieda wine were collected to establish chemical constituents database. The structure of the compounds was identified on the basis of above constituents database ,the relevant literature ,retention time of reference substance and MS fragmentation regularity. RESULTS & CONCLUSIONS : Fifty-nine compounds were identified in Fengliaoxing fengshi dieda wine ,including 12 flavones(e.g. neoeriocitrin ,hesperidin.),8 alkaloids (e.g. baogongteng C or erycibellin , ephedrine,pseudoephedrine), 9 organic acids (e.g. chlorogenic acid , cryptochlorogenic acid ,neochlorogenic acid ),7 coumarins(e.g. xanthotoxol), 4 esters, 4 amino acids , and 15 other categories(including volatile oils ,terpenes,amides). UPLC- 5103YX11SF37(17)] Q-Exactive-MS technology can quickly and accurately identify the chemical constituents in Fengliaoxing fengshi dieda wine.

13.
Artigo em Chinês | WPRIM | ID: wpr-865285

RESUMO

Objective:To compare the tropism of different adeno-associated virus (AAV) serotypes in retinal cells.Methods:The plasmids pFastBacDual-inCap and pFastBacDual-ITR-CMV-EGFP were constructed for AAV packaging with the baculovirus expression system.Recombinant adeno-associated virus type 2(rAAV2), 6, 8 and 9 serotypes were packaged, and the infectivity of rAAV was evaluated by infecting HEK293T cells at multiplicity of infection(MOI)2000.Twenty-five C57BL/6 mice were divided into five groups, with five mice per group.In the three experimental groups, both eyes of each mouse were injected 1 μl rAAV intravitreally, and 1 μl phosphate buffered saline (PBS) for the eyes of the control group.Two weeks after injection, the retinal tissues were collected for preparing flat mounts and cryosections.Enhanced green fluorescent protein (EGFP) gene expression was observed via fluorescence microscopy and laser scanning confocal microscopy.The study protocol was approved by the Ethics Committee of Suzhou Institute of Biomedical Engineering and Technology.Results:The infection efficiency of the recombinant virus to HEK293T cells was rAAV2>rAAV6>rAAV8>rAAV9, and the transduction efficiency was 39.5%, 18.4%, 8.7% and 4.6%, respectively.In mouse retinal transduction, rAAV2 and rAAV6 were highly expressed in the ganglion cells, and rAAV8 and rAAV9 were highly expressed in the retinal pigment epithelium (RPE) and photoreceptor cells.rAAV2-mediated EGFP expression in retinas was stable within three months after injection.Conclusions:Different rAAV serotypes have varying tropism and transduction efficiencies in retinal cells through intravitreal injection, rAAV2 has a high transduction efficiency and it can be stably expressed in retinas within three months after injection.

14.
Artigo em Chinês | WPRIM | ID: wpr-871354

RESUMO

Objective:To investigate the regulatory effects of sphingosine kinase-2 (SphK2) on the function of activated astrocytes and the progression of experimental autoimmune encephalomyelitis (EAE) in mice.Methods:Primary mouse astrocytes were isolated from wild-type (WT) C57BL/6 mice and sphk2 gene knock-out ( sphk2 -/-) mice and stimulated in vitro with interleukin 17 (IL-17). Real-time PCR was used to measure the expression of inflammatory cytokines and chemokines at mRNA levels. Western blot and immunofluorescence were used to detect the expression of glial fibrillary acidic protein (GFAP) and the phosphorylation of signal transducer and activator of transcription 3 (p-STAT3). An EAE mouse model was constructed using myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55) polypeptide. Western blot was used to detect the expression of GFAP and p-STAT3 at protein level and real-time PCR was used to detect the expression of inflammatory cytokines and chemokines at mRNA level in spinal cords. Hematoxylin-Eosin (HE) and Luxol fast blue (LFB) staining were used to observe the changes in inflammatory cell infiltration and demyelination in spinal cords. Results:Compared with the WT group, the phosphorylation of STAT3 was obviously reduced in in vitro activated mouse astrocytes of sphk2 -/- mice, and the expression of inflammatory cytokines and chemokines including monocyte chemotactic protein 1 (MCP-1), TNF-α and IL-6 at mRNA level was also significantly decreased. Compared with the WT EAE group, changes in the above-mentioned cytokines and relative proteins in sphk2 -/- EAE mice in vivo were similar to those in vitro. Moreover, inflammatory cell infiltration and demyelination were significantly reduced in spinal cords of sphk2 -/- EAE mice. However, no significant difference in in vitro or in vivo GFAP expression was observed between WT and sphk2 -/- mice. Conclusions:SphK2 might regulate the function of reactive astrocytes through STAT3 molecular pathway, thereby regulating the production of inflammatory cytokines and chemokines and participating in the pathological process of EAE.

15.
Artigo em Chinês | WPRIM | ID: wpr-797459

RESUMO

Objective@#To investigate the current development status quo of child healthcare by analyzing Chinese literatures on pediatric nursing teaching published over the recent five years via co-word analyzing method.@*Methods@#Literatures published between 2013-2017 and had subject headings including "pediatrics", "nursing" and "teaching" were retrieved from the Chinese National Knowledge Integrated Database, VIP website and Wanfang Data; all included literatures were analyzed via Bicomb 2; keyword co-occurrence diagram was drawn using Ucinet 6.0 and Netdraw to visualize the co-occurrence relations among high-frequent words.@*Results@#A total of 396 literatures were included and 29 high-frequency keywords were extracted. Results showed that studies on pediatric nursing teaching in China focused on the teaching mode reform, especially PBL teaching method, CBL teaching method, situational simulation teaching method, clinical pathway teaching method, etc.; literatures on undergraduates, higher vocational education and teaching quality evaluation were more than those on secondary vocational education, experimental teaching and humanistic care.@*Conclusion@#Researches about pediatric nursing have made some progress in teaching reform, teaching quality evaluation and undergraduate education in China. In the future, we can further explore appropriate teaching mode for each part, and attach more attention to researches on secondary vocational education, experimental teaching and humanistic care teaching.

16.
Artigo em Chinês | WPRIM | ID: wpr-790262

RESUMO

Objective To investigate the current development status quo of child healthcare by analyzing Chinese literatures on pediatric nursing teaching published over the recent five years via co-word analyzing method. Methods Literatures published between 2013-2017 and had subject headings including "pediatrics", "nursing" and "teaching" were retrieved from the Chinese National Knowledge Integrated Database, VIP website and Wanfang Data; all included literatures were analyzed via Bicomb 2; keyword co-occurrence diagram was drawn using Ucinet 6.0 and Netdraw to visualize the co-occurrence relations among high-frequent words . Results A total of 396 literatures were included and 29 high-frequency keywords were extracted. Results showed that studies on pediatric nursing teaching in China focused on the teaching mode reform, especially PBL teaching method, CBL teaching method, situational simulation teaching method, clinical pathway teaching method, etc.; literatures on undergraduates, higher vocational education and teaching quality evaluation were more than those on secondary vocational education, experimental teaching and humanistic care. Conclusion Researches about pediatric nursing have made some progress in teaching reform, teaching quality evaluation and undergraduate education in China. In the future, we can further explore appropriate teaching mode for each part, and attach more attention to researches on secondary vocational education , experimental teaching and humanistic care teaching.

17.
Artigo em Chinês | WPRIM | ID: wpr-756612

RESUMO

Early intervention contributes to improving patient experience and doctor-patient relationship in the case of non-psychiatric outpatients with psychological problems.The authors studied the psychological assessment and hierarchical management for non-psychiatric inpatients at a general hospital. Measures taken include establishing multi-disciplinary and inter-department teams, building an intra-hospital joint-action system, and implementing the psychological assessment and hierarchical management for non-psychiatric inpatients.These efforts explored ways for a general hospital in psychological counseling, offering humanistic service, and transformation of medical pattern.

18.
Artigo em Chinês | WPRIM | ID: wpr-693136

RESUMO

Cervical cancer is a common malignant tumor of women,which is caused by human papillomavirus(HPV) infection especially high risk HPV infection(HR-HPV).According to the statistics from the World Health Organization (WHO),the incidence and fatality rate of cervical cancers are obviously rising and getting younger in resent years,which seriously threatens the health of women.In China,the mortality of cervical cancer is the first in the female reproductive tract malignant tumor.Although cervical cancer screening has received more and more attention,which reduces the risk of cervical cancer,it does not prevent the necessary factors of cervical cancer,i.e.the HPV infection.Therefore,effective prevention and treatment methods are urgently needed to control the occurrence and development of HPV infection.Both Chinese and Western medicine have made significant progress in the treatment of persistent HPV infection,and they have own advantages and can complement each other.The combination of Chinese and Western medicine is expected to be an effective way to treat cervical persistent HPV infection.

19.
Artigo em Chinês | WPRIM | ID: wpr-752037

RESUMO

"If slight diarrhoea, replacing Ephedra with Daphninae"under the article 40 of Treatise on Febrile Diseasesis is controversial. There are two different opions in all physicians, one explaination it is in line with Zhang Zhongjing but without the evidence about"replacing Ephedra with Daphninae"; the other is that Daphninae is too strong to apply in diarrhoea. According to the analysis on Treatise on Febrile and Miscellaneous Diseases, the slight diarrhoea is the phenomenon of Vital qi exorcism. The reason why replacing Ephedra owing to exogenous evil disappearance, the construction of Daphninae Pinellia ternate and Paeony is to expell the water on the leaving way. And we draw a conclusion that the function of Paeony in Xiaoqinglong decoction is inclined to be expelling Water Qi.

20.
Artigo em Chinês | WPRIM | ID: wpr-696593

RESUMO

Nocturnal enuresis is a common disease in pediatrics.The pathophysiology of enuresis is complex,involving nocturnal polyuria,overactive bladder,abnormal nocturnal plasma level of vasopressin,and genetic factors.Its treatments include basic treatment,conditioning alarm regimen,pharmacotherapy,and psychotherapy.This article reviews the recent advances in the pathogenesis,diagnosis,and treatment of enuresis and aims to provide references for the diagnosis and management of children with nocturnal enuresis.

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