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1.
Acta Academiae Medicinae Sinicae ; (6): 16-21, 2023.
Article in Chinese | WPRIM | ID: wpr-970440

ABSTRACT

Objective To investigate the role and mechanism of circ_0092315 in the proliferation and invasion of papillary thyroid carcinoma cells. Methods The expression of circ_0092315 in papillary thyroid carcinoma cells was examined by real-time fluorescence quantitative PCR.The proliferation and invasion of TPC-1 cells was assessed by CCK-8 and Transwell assays.The protein level of high mobility group A2 (HMGA2) was determined by Western blotting.The regulatory relationship of circ_0092315,microRNA-1256 (miR-1256),and HMGA2 was explored by bioinformatics tools,dual-luciferase reporter assay,real-time fluorescence quantitative PCR,and Western blotting. ++++Results circ_0092315 was overexpressed in papillary thyroid carcinoma cells (all P<0.001).circ_0092315 promoted the proliferation and invasion of TPC-1 cells (all P<0.001).The transfection of si-circ_0092315 up-regulated the expression of miR-1256 (P<0.001),and miR-1256 inhibitor up-regulated the protein level of HMGA2 (P<0.001). ++++Conclusion circ_0092315 is overexpressed in TPC-1 cells and it promotes the proliferation and invasion of TPC-1 cells by regulating the miR-1256/HMGA2 axis.


Subject(s)
Humans , Thyroid Cancer, Papillary/genetics , Computational Biology , Thyroid Neoplasms/genetics , Cell Proliferation , MicroRNAs/genetics
2.
Chinese Acupuncture & Moxibustion ; (12): 559-562, 2022.
Article in Chinese | WPRIM | ID: wpr-927424

ABSTRACT

"Unblocking fu organs" is one of the essential principles of Ma's warm moxibustion technique, characterized as "dredging" and "harmonizing" for either deficiency or excess condition. Under the guidance of this therapeutic thought, the acupoints for moxibustion are mainly selected from the middle and lower parts of the body. Regarding the therapeutic approach, the acupoint prescription for moxibustion should be formed in line with warming and promoting circulation of fu organs; the moxibustion degree should be specially considered, in which, the mild moxibustion is recommended to induce promoting action; and the systematic moxibustion technique should be the root for dredging fu organs and regulating zang organs. Ma's mild moxibustion technique stresses on removing the obstruction of fu organs and emphasizes promoting the qi activity of sanjiao (triple energizer) and regulating the balance of five zang organs.


Subject(s)
Humans , Acupuncture Points , Acupuncture Therapy , Ethnicity , Hyperplasia , Moxibustion/methods
3.
Chinese Journal of General Practitioners ; (6): 50-54, 2021.
Article in Chinese | WPRIM | ID: wpr-885330

ABSTRACT

Objective:To investigate the current status of chronic respiratory disease (chronic obstructive pulmary disease, bronchial asthma and obstructive sleep apnea) management capacity in community health centers in Beijing Miyun district.Methods:From November 21 to 22,2018,nineteen community health centers and 65 general practitioners in Miyun district of Beijing were selected to participate in a questionnaire survey. The self-designed questionnaire was divided into two parts: the questionnaire for medical institutions(institution questionnaire)and questionnaire for general practitioners(doctor questionnaire). The institution questionnaires were distributed by the Miyun District Health Commission,and filled in by the person in charge of the institution; the knowledge questionnaires were sent to all general practitioners of 4 community health service centers. The two independent sample t test was used to compare the measurement data in accordance with normal distribution between the two groups, and analysis of variance was used for multi group comparison. Results:Nineteen institution questionnaires were sent to all centers in the district and all 19 valid questionnaires were recovered. Among them, 18 centers thought that chronic respiratory diseases should be included in the management of chronic non-communicable diseases, and health records should be established to achieve regular follow-up monitoring, but only one center had put asthma in the record. Nine centers purchased pulmonary function instrument; 8 centers were equipped with inhaled glucocorticoid, and 1 center was equipped with β 2-receptor agonists. The effective recovery rate of knowledge questionnaire was 100.0% (65/65). There was no significant difference in the knowledge scores of three kinds of chronic respiratory diseases (chronic obstructive pulmary disease, bronchial asthma, obstructive sleep apnea) among general practitioners [(63±19), (64±23), (62±21), F=0.087, P>0.05]. The scores of general practitioners with different ages and professional titles were (57±15), (66±13), (42±16) and (54±19), (67±12), (68±11) respectively. There were significant differences in the knowledge scores of general practitioners with different ages and professional titles ( F= 8.582 and 6.079, all P<0.05). The average scores of general practitioners with age>50 years or junior professional title were lower than others. Conclusions:The leaders of each center in Miyun district have a clear understanding of the necessity of diagnosis and treatment of chronic respiratory diseases, but there are still some problems, such as insufficient attention to chronic respiratory system, insufficient investment in disease management infrastructure, and lack of professional knowledge of chronic respiratory diseases among general practitioners. It is hoped that in the future, chronic respiratory diseases can be introduced into public health service projects, investment in related disease infrastructure will be strengthened, and comprehensive respiratory knowledge and ability training courses suitable for grass-roots general practitioners can be popularized as soon as possible.

4.
Chinese Acupuncture & Moxibustion ; (12): 1349-1353, 2021.
Article in Chinese | WPRIM | ID: wpr-921058

ABSTRACT

OBJECTIVE@#To observe the efficacy of acupoint thread-embedding combined with fluticasone propionate aerosol for chronic persistent bronchial asthma of lung-spleen @*METHODS@#A total of 120 children with chronic persistent bronchial asthma were randomly divided into an observation group (60 cases, 9 cases dropped off) and a control group (60 cases, 7 cases dropped off). The control group was treated with fluticasone propionate aerosol (125 μg per inhalation), twice a day; based on the control group treatment, the observation group was treated with acupoint thread-embedding at Dingchuan (EX-B 1), Feishu (BL 13), Zusanli (ST 36) and Danzhong (CV 17), once half a month. Both groups were treated for 3 months. The pulmonary function, serum IgA, IgE levels and TCM symptom score were compared between the two groups before and after treatment, and the clinical efficacy was evaluated.@*RESULTS@#After treatment, the large airway function (peak expiratory flow [PEF], forced expiratory volume at the first second [FEV1]) and small airway function (maximal expiratory flow at 25% of the forced capacity [MEF25%], maximal expiratory flow at 50% of the forced capacity [MEF50%], maximal expiratory flow at 75% of the forced capacity [MEF75%] and midexpiratory flow 25%-75% [MEF25%-75%]) were higher than those before treatment (@*CONCLUSION@#Acupoint thread-embedding combined with fluticasone propionate aerosol could improve the pulmonary function, TCM symptoms and serum IgA and IgE levels in children with chronic persistent bronchial asthma of lung-spleen


Subject(s)
Child , Humans , Acupuncture Points , Acupuncture Therapy , Asthma/drug therapy , Immunoglobulin A , Immunoglobulin E , Lung
5.
Journal of Experimental Hematology ; (6): 2033-2038, 2020.
Article in Chinese | WPRIM | ID: wpr-880011

ABSTRACT

OBJECTIVE@#To analyze the molecular pathogenesis by analysis of phenotype and gene mutation in families with hereditary coagulation factor V (FⅤ) defect caused by complex heterozygous mutation.@*METHODS@#Plasma pro-thrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), FⅤ procoagulant activity (FⅤ∶C), FⅤ antigen (FⅤ∶Ag), and other related coagulation indexes were detected in the proband and his family members (3 generations 10 people). Using DNA direct sequencing to analyze all exons, flanks, 5' and 3' untranslated regions of F5 genes and the corresponding mutation site regions of family members, the mutation site was confirmed by reverse sequencing.The conservation of mutant amino acids was analyzed by ClustalX-2.1-win software. The PROVEAN and MutationTaster online bioinformatics software were used to predict the effect of mutation on protein function. Protein model and amino acid interaction at mutation sites was analyzed by Swiss-pdbviewer software.@*RESULTS@#The PT and APTT of the proband were significantly prolonged compared with healthy controls (34.2 vs 13.2 s and 119.3 vs 36.0 s), while FⅤ∶C and FⅤ∶Ag extremely reduced (3% and 6%). The PT and APTT of the second-born, the third son, daughter, and grandson of the proband were slightly prolonged, and the FⅤ∶C and FⅤ∶Ag decreased to varying degrees. The related coagulant parameters of other family members were within normal range. Genetic analysis revealed that the proband had a c.911G>A heterozygous missense mutation on the exon 6 lead to p.Gly276Glu, and a c.5343C>G heterozygous missense mutation on the exon 16 lead to p.Ser1781Arg of the proband. The second-born, the third son, and grandson of the proband carry p.Gly276Glu heterozygotes, and the daughter carries p.Ser1781Arg heterozygotes, while the other family members were wild-type. The results of conservative analysis indicated that p.Gly276 and p.Ser1781 were highly conserved in homologous species. The two bioinformatics software predicted the same results, PROVEAN (score -6.214 and -12.79) indicated that the compound heterozygous mutation was a harmful mutation; MutationTaster (score 0.976 and 0.999) suggested that these mutations might cause corresponding disease. p.Gly276Glu protein model analysis showed that, the Glu side chain was prolonged and the molecular weight became larger, which would increase the steric hindrance between it and the surrounding amino acids, affect the normal local folding of the FⅤ protein, and eventually lead to the decrease of protein activity and content. This paper can not provide analysis of the spatial structure of p.Ser1781Arg mutant protein because of the lack of X ray 3 D structure file of FⅤ exon 16.@*CONCLUSION@#The new compound heterozygous mutations (p.Gly276Glu and p.Ser1781Arg) identified in this study are the main reasons for the decrease in the FⅤ level of the family, among which p.Ser1781Arg is rarely reported at home and abroad.


Subject(s)
Humans , Factor V/genetics , Family , Genotype , Heterozygote , Mutation , Pedigree , Phenotype
6.
Chinese Journal of Applied Physiology ; (6): 308-311, 2019.
Article in Chinese | WPRIM | ID: wpr-776508

ABSTRACT

OBJECTIVE@#To observe the expressions of sphingosine kinase 1 (SphK1) and sphingosine-1-phosphate receptor 2 (S1PR2) in hippocampus of epileptic rats and to investigate the pathogenesis of SphK1 and S1PR2 in epilepsy.@*METHODS@#One hundred and eight male Sprague-Dawley (SD) rats were randomly divided into control group (n=48) and pilocarpine (PILO) group (n=60). A robust convulsive status epilepticus (SE) was induced in PILO group rats by the application of pilocarpine. Control group rats were injected with respective of physiological saline. Pilocarpine group was randomly divided into 6 subgroups (n=8): acute group (E6 h, E1 d, E3 d), latent group (E7 d) and chronic group (E30 d, E56 d). Each subgroup has 8 control rats and 8 epileptic rats. Hippocampal tissue and brain slices were obtained from control rats and rats subjected to the Li-PILO model of epilepsy at 6 h, 1 d, 3 d,7 d,30 d and 56 d after status epilepticus (SE). Western blot technique was used to determine the expressions of SphK1 and S1PR2 in hippocampus at different point of time after pilocarpine treatment. Immunofluorescence was applied to detect the activation and proliferation of hippocampal astrocytes and the localization of SphK1 and S1PR2 in rat hippocampal astrocytes.@*RESULTS@#Compared with control group, the levels of SphK1 in acute phase (E3 d), latent phase (E7 d) and chronic phase (E30 d, E56 d) were significantly increased while the expressions of S1PR2 were decreased in acute phase (E3 d), latent phase (E7 d) and chronic phase (E30 d, E56 d)(P<0.05 or P<0.01). Immunofluorescence results showed astrocyte activation and proliferation in hippocampus of epileptic (E7 d) rats (P<0.05). Confocal microscopy confirmed the preferential expressions of SphK1 and S1PR2 in epileptic rat(E7 d)hippocampal astrocytes.@*CONCLUSION@#The results indicate that SphK1 and S1PR2 may play an important role in the pathogenesis of epilepsy by regulating the activation and proliferation of hippocampal astrocytes and altering neuronal excitability.


Subject(s)
Animals , Male , Rats , Astrocytes , Epilepsy , Hippocampus , Cell Biology , Phosphotransferases (Alcohol Group Acceptor) , Metabolism , Pilocarpine , Random Allocation , Rats, Sprague-Dawley , Receptors, Lysosphingolipid , Metabolism
7.
Chinese Journal of Medical Education Research ; (12): 286-290, 2019.
Article in Chinese | WPRIM | ID: wpr-744172

ABSTRACT

The National Medical Licensing Examination has become one of the most important indicator s to measure the teaching quality of medical colleges and universities. In this paper, by analyzing the status of pathophysiology in National Medical Licensing Examination and the current problems existing in pathophysiology teaching, the author proposed a scheme of reform in the pathophysiology teaching based on Medical Licensing Examination, including changing teaching idea, optimizing teaching content, reforming teaching means and adjusting the assessment methods . This reform aims to make the pathophysiology teaching really serve the needs of clinical application.

8.
Chinese Journal of Applied Physiology ; (6): 102-105, 2018.
Article in Chinese | WPRIM | ID: wpr-773794

ABSTRACT

OBJECTIVES@#To investigate the role of autophagy inhibitor chloroquine (CQ) in acute ethanol-induced liver injury and its mechenism.@*METHODS@#Twenty-one C57BL/6 male mice were randomly divided into three groups:control group, ethanol group, CQ + ethanol group (=7). Mice in ethanol group were administered 33% (v/v) ethanol at a dose of 4.5 g/kg body weight. Ethanol-induced liver steatosis in each group was detected by hematoxylin and eosin staining. Hepatic lipid accumulation was detected by staining with Oil red O. Hepatic tissue triglyceride (TG) levels, serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) were determined by biochemical assays. Protein expression of microtubule-associated protein 1 light chain 3(LC3) and nuclear factorκB p65(NF-κB p65) were measured by Western blot and immunofluorescence. Pro-inflammatory factors tumor necrosis factor-α(TNF-α)、interleukin 6(IL-6) were detected by ELISA.@*RESULTS@#Compared with control group, ethanol induced liver injury proved by accumulation of hepatic lipids, TG levels, AST and ALT activities were significantly increased by ethanol, protein expression of LC3-Ⅱ was also markedly increased by ethanol. Compared with ethanol group, addition of CQ increased furtherthe level of LC3-Ⅱexpression, and TG amount, serum AST and ALT activities, and the expression of NF-κB p65, TNF-αand IL-6.@*CONCLUSIONS@#Acute ethanol-intake could induce liver steatosis and inflammation, and autophagy inhibitor CQ exacerbatedethanol-induced liver injury, suggested that autophagy might be protective effect in acute ethanol-induced liver disease.


Subject(s)
Animals , Male , Mice , Alanine Transaminase , Blood , Aspartate Aminotransferases , Blood , Autophagy , Chloroquine , Pharmacology , Interleukin-6 , Liver , Liver Diseases, Alcoholic , Drug Therapy , Mice, Inbred C57BL , Microtubule-Associated Proteins , Metabolism , Random Allocation , Transcription Factor RelA , Metabolism , Triglycerides , Tumor Necrosis Factor-alpha
9.
Chinese Journal of Emergency Medicine ; (12): 394-398, 2018.
Article in Chinese | WPRIM | ID: wpr-694391

ABSTRACT

Objective To study the predictive value of serum soluble ST2 (sST2) in patients with sepsis.Methods A total of 63 patientswith sepsis and 30 healthy subjects as a control group in the emergency Department,Beijing Hospital,National Center of Gerontology,were enrolled in the study.Serum sST2 concentrations were measured by ELISA method.Patients were divided into sepsis group (n=44) and septic shock group (n=19).According to 28-day mortality after the diagnosis of sepsis,patients were divided into death group (n=18) and survival group (n=45).Respiratory rate,oxygenation index,white blood cell count,procalcitonin (PCT),C reactive protein (CRP),serum creatinine (CRE),total bilirubin (TBIL) of patients and control subjects were measured.SPSS 23.0 software was used for the statistical analyses.The measurement data was analyzed by t test and the enumeration data was analyzed by Chi square test.The survival status was analyzed by Logistic binary regression analysis and ROC curve analysis.Results The serum sST2 level (1 382.12±384.07) pg/mL in sepsis group was significantly higher than that in control group (569.28±163.46) pg/mL (P<0.05).in septic shock group,28-day mortality rate (63.16%) and serum sST2 level (1 675.49±457.59) pg/mL was higher than those in sepsis group (13.64%) (1255.44 ± 265.70) pg/ml (P<0.05).The PCT (16.37±16.36) ng/mL and serum sST2 level (1794.47±335.18)pg/mL in death group were higher than those in survival group (P<0.05).The ROC curve showed that the AUC of sST2 was larger than that of PCT (0.917 vs.0.884),the sensitivity was higher than that of PCT (88.9% vs.72.2%),and the specificity was lower than that of PCT (82.2% vs.93.3%).The combination AUC of sST2 and PCT was 0.944.Conclusions Serum sST2 has a certain value in the diagnosis of sepsis,and can be used to predict the prognosis of patients with sepsis.The higher the sST2 value,the worse the prognosis.Compared with PCT,sST2 is more sensitive in the prognosis of sepsis,but the specificity is not high enouph.The measurement of sST2 level coupled with PCT level may be more useful.

10.
China Journal of Chinese Materia Medica ; (24): 3740-3747, 2018.
Article in Chinese | WPRIM | ID: wpr-689852

ABSTRACT

To study the effect of polysaccharides from Polygonatum sibiricum on mRNA and protein expressions of blood lipid metabolism in hyperlipidemic mice. The mice were randomly divided into 6 groups, namely the blank control group, the hyperlipidemia model group, the simvastatin group, and low, middle and high-dose PSP groups (200, 400, 800 mg·kg⁻¹·d⁻¹). Each group of the mice was administrated intragastrically for 14 days, respectively. Subsequently, every group of mice, except for the blank control group, was intraperitoneally injected with 75% fresh egg yolk emulsion for establishing the hyperlipidemic mice model. Upon completion of the administration, the contents of TC, TG, LDL-C and HDL-C in serum of each group were investigated in details. In particular, the mRNA expression levels of PPAR-α, PPAR-, PPAR-, SREBP-1c, IL-6 and TNF-α of the liver tissues were detected by Real-time PCR, and the protein expression levels (including PPAR-α, PPAR-, PPAR-, SREBP-1c, IL-6, TNF-α) were examined by Western blot. Consequently, the obtained results showed that the contents of the serum TC, TG, LDL-C of low, middle and high-dose PSP groups significantly decreased compared with those of the hyperlipidemia model group. Simultaneously, there were significant differences between middle-dose and high-dose PSP groups (<0.01). In striking contrast, the contents of serum HDL-C of low, middle and high-dose PSP groups significantly increased, while obvious differences were also observed between middle-dose and high-dose PSP groups (<0.01). Moreover, middle-dose and high-dose PSR groups could up-regulate the protein and mRNA expressions of PPAR-α, PPAR- (<0.05) compared with those of the hyperlipidemia model group, and down-regulate the expressions of PPAR-,SREBP-1c, IL-6 and TNF-α(<0.05) compared with those of liver tissues of the hyperlipidemia model group. In conclusion, all of the above results suggested that PSP could inhibit the oxidation of the liver lipid, and regulate the expression levels of the corresponding genes and proteins relating to the lipid metabolism, so as to play a critical role for preventing hyperlipidemia.

11.
Chinese Medical Journal ; (24): 93-99, 2017.
Article in English | WPRIM | ID: wpr-303196

ABSTRACT

<p><b>BACKGROUND</b>Epigallocatechin-3-gallate (EGCG) has exhibited antitumor properties in several types of cancers, including nasopharyngeal carcinoma (NPC), but the molecular mechanisms underlying this function remain incompletely understood. The aim of the present study was to characterize the global impact of EGCG on the expression of microRNAs (miRNAs) in NPC cells.</p><p><b>METHODS</b>Using microarray analysis, the alterations of miRNA expression profiles were investigated in EGCG-treated CNE2 cells. Furthermore, the target genes and signaling pathways regulated by EGCG-specific miRNAs were identified using target prediction program and gene ontology analysis.</p><p><b>RESULTS</b>A total of 14 miRNAs exhibited >2-fold expression changes in a dose-dependent manner after treatment with 20 μmol/L and 40 μmol/L EGCG. Totally 43, 49, and 52 target genes from these differentially expressed miRNAs were associated with the apoptosis, cell cycle regulation, and cell proliferation, respectively. A total of 66 signaling pathways, primarily involved in cancer development and lipid and glucose metabolism, were shown to be regulated by EGCG-specific miRNAs.</p><p><b>CONCLUSION</b>EGCG induces considerable alterations of miRNA expression profiles in CNE2 cells, which provides mechanistic insights into cellular responses and antitumor activity mediated by EGCG.</p>


Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Carcinoma , Catechin , Pharmacology , Cell Line, Tumor , Computational Biology , Gene Expression , Genetics , MicroRNAs , Genetics , Metabolism , Nasopharyngeal Neoplasms , Genetics , Real-Time Polymerase Chain Reaction , Signal Transduction , Genetics
12.
Journal of Experimental Hematology ; (6): 1753-1757, 2015.
Article in Chinese | WPRIM | ID: wpr-272527

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the hematopoietic reconstitution in immunodeficiency NPG(TM) mice after transplantation of G-CSF-mobilized peripheral blood CD34(+) hemopoietic stem cells.</p><p><b>METHODS</b>CD34(+) cells were isolated from peripheral blood stem cells (PBSC) by magnetic activated cell sorting (MACS), and then were transplanted into NPG(TM) mice irradiated with sublethal dose of X ray by marrow cavity transplantation. The hemogram of mice after transplantation for 2, 4 weeks was observed; human cell populations (CD45(+), CD19(+)) in the peripheral blood of mice were dynamically analyzed by flow cytometry (FCM) at 4, 6, 8, 10 and 12 weeks after transplantation. Until the planned harvest at the 12 week after transplantation, the CD45(+), CD19(+) level in bone marrow, liver, spleen from each mouse were detected by flow cytometry; the expression of human Alu gene in the bone marrow cell of mouse was detected by PCR.</p><p><b>RESULTS</b>The purity of CD34(+) cells accounted for 96.3%; after irradiation, the nucleated cells and megalokaryocytes in the marrow cavity of NPG mice were reduced significantly or were lost, and reached the myeloablative effect. At week 4 after transplantation, components of blood cells in peripheral blood of transplanted mice were recovered to the level before irradiation; all the mice survived, human CD45(+), CD19(+) cells were found by FCM in the peripheral blood of all the surviving mice in transplantation group at week 4, 6, 8, 10, 12 after the transplantation; at the 12th week, the human Alu gene could be detected in the bone marrow of all the mice in transplantation group.</p><p><b>CONCLUSION</b>The human-mouse chimeric model is successfully established in irradiation-induced NPG mouse by transplantation of CD34(+) HSC from G-CSF-mobilized peripheral blood via marrow cavity.</p>


Subject(s)
Animals , Humans , Mice , Bone Marrow , Bone Marrow Cells , Bone Marrow Transplantation , Cord Blood Stem Cell Transplantation , Disease Models, Animal , Granulocyte Colony-Stimulating Factor , Hematopoietic Stem Cells , Spleen
13.
Chinese Journal of Pathophysiology ; (12): 1557-1562, 2015.
Article in Chinese | WPRIM | ID: wpr-479293

ABSTRACT

AIM:To study the effect of epigallocatechin-3-gallate (EGCG) on the proliferation of human naso-pharyngeal carcinoma ( NPC) cells, and to explore its mechanism by targeting miR-34a.METHODS: Nasopharyngeal carcinoma CNE-2Z cells were treated with various concentrations of EGCG .The ability of cell proliferation was detected by CCK-8 assay, 5-ethynyl-2-deoxyuridine (EdU) incorporation assay and colony-forming assay.The cell cycle distributions were analyzed by flow cytometry .The protein levels of P53 and Notch1 were detected by Western blot .The expression of miR-34a and Notch1 mRNA was measured by real-time PCR.RESULTS:EGCG effectively inhibited the proliferation and colony formation of CNE-2Z cells in a dose-dependent manner , which was related to its induction of cell cycle arrest at G 0/G1 phase.The expression of P53 and miR-34a in CNE-2Z cells was significantly increased after treated with EGCG , while the expression of Notch1 at mRNA and protein levels was markedly suppressed .CONCLUSION:EGCG induces cell cycle arrest and suppresses cell proliferation by regulating the P 53/miR-34a/Notch1 pathway in NPC cells.

14.
Chinese Journal of Tissue Engineering Research ; (53): 1488-1494, 2015.
Article in Chinese | WPRIM | ID: wpr-465372

ABSTRACT

BACKGROUND: Cyclooxygenase 2, aggrecanase 1, and insulin-like growth factor 1 are involved in pathological injury of the articular cartilage. OBJECTIVE:To observe the expression of shRNA vectors carrying cyclooxygenase 2, aggrecanase 1 and overexpression vectors carrying insulin-like growth factor 1 in bone marrow mesenchymal stem cels. METHODS:Lentiviral vectors carrying the silencing gene cyclooxygenase 2, aggrecanase 1, the over-expressing gene insulin-like growth factor 1 and binding green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect passage 3 human bone marrow mesenchymal stem cels culturedin vitro (experimental group). The human bone marrow mesenchymal stem cels transfected with no target gene lentivirals were used as negative control group. The human bone marrow mesenchymal stem cels transfected with no treatment served as blank group. RESULTS AND CONCLUSION:Cyclooxygenase 2 and aggrecanase 1 transfected in human bone marrow mesenchymal stem cels were significantly inhibited at gene and protein levels, while the expression of insulin-like growth factor 1 was increased significantly at gene and protein levels. We confirmed that cyclooxygenase 2 and aggrecanase 1 were successfuly silenced while insulin-like growth factor 1 overexpressed by using lentiviral vectors in human bone marrow mesenchymal stem cels, which brings a new hope for the systemic gene treatment of arthritis.

15.
Chinese Journal of Applied Physiology ; (6): 8-12, 2014.
Article in Chinese | WPRIM | ID: wpr-235309

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of autophagy inhibitor chloroquine (CQ) in the proliferation of pulmonary arterial smooth muscle cells (PASMCs) in hypoxia conditions.</p><p><b>METHODS</b>The following groups in this study were set up: control group, hypoxia group, 50 micromol/L CQ + hypoxia group, 50 micromol/L CQ group. The viability of PASMCs in every group was detected by MTT assay. Autophagic vacuoles in the cells were observed by MDC staining. Protein expression of microtubule associated protein light chain 3 (LC3) was measured by Western blot. Migration of PASMCs was detected by wound healing assay.</p><p><b>RESULTS</b>Compared with control group, no effect on the viability of PASMCs was observed treated by CQ alone. In 1% hypoxia group, cell viability increased significantly compared with that in control group. The number of autophagic vacuoles and the rate of cell migration and also protein expression of LC3-II were also markedly increased. Compared with hypoxia group, addition of CQ increased the number of autophagic vacuoles and the levels of LC3-II protein, but decreased the proliferation and migration of PASMCs.</p><p><b>CONCLUSION</b>Hypoxia could activates autophagy and contributes to proliferation and migration of PASMCs, and autophagy inhibitor CQ could decrease the effect of hypoxia on PASMCs through inhibiting autophagy process.</p>


Subject(s)
Humans , Autophagy , Cell Hypoxia , Cell Movement , Cell Survival , Cells, Cultured , Chloroquine , Pharmacology , Microtubule-Associated Proteins , Metabolism , Myocytes, Smooth Muscle , Pulmonary Artery , Cell Biology
16.
Chinese Journal of Industrial Hygiene and Occupational Diseases ; (12): 522-525, 2013.
Article in Chinese | WPRIM | ID: wpr-343613

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the protective effect of suppressive oligodeoxynucleotides (Sup ODN) on interferon-γ (IFN-γ) and signal transducers and activators of transcription (pSTAT4) expression of Silica-induced pulmonary inflammation in Mice.</p><p><b>METHODS</b>Sixty Balb/c mice were randomly divided into 4 groups, normal control group, silicious group, suppressive oligodeoxynucleotides (Sup ODN) group, control oligodeoxynucleotides (Con ODN) group. Except the normal control group injected normal saline, the rest groups were induced by the intratracheal instillation of 0.1 ml (5 g/L) of sterilized silica suspension. Sup ODN group and Con ODN group were treated by i.p. injection of 0.3 ml (1mg/mL) of suppressive or control ODN 3 h before silica administration. After 7 days, the animals were killed and levels of IFN-γ were detected by ELISA. The pathologic changes in lung tissues of mice were observed with HE staining. Expressions of IFN-γ and pSTAT4 in lung tissue were detected with immunohistochemistry and quantified by Image-Pro Plus 7.0.</p><p><b>RESULTS</b>HE staining showed that the lung tissue of silicious group were damaged seriously than Sup ODN group. Compared with the normal control group (serum: (280.1±41.3) pg/ml, lung tissue: (0.249±0.373), IFN-γ increased in silicious group (serum: (886.3±81.7) pg/ml, lung tissue: (0.270±0.300) (P < 0.05). Compared with the normal control group and Con ODN group [(894.5±91.6) pg/ml], IFN-γ in the serum of Sup ODN group decreased significantly (P < 0.01). Compared with the silicious group , IFN-γ in lung tissue decreased in Sup ODN group (0.241±0.250) (P < 0.05). Compared with the normal control group (0.279±0.353), pSTAT4 in lung tissue increased significantly in silicious group (0.313±0.231) (P < 0.01). Compared with the silicious group, pSTAT4 in lung tissue decreased significantly in Sup ODN group (0.269±0.523) (P < 0.01).</p><p><b>CONCLUSION</b>Sup ODN attained protective effect on Silica treated mice by suppressing expression of IFN-γ and pSTAT4.</p>


Subject(s)
Animals , Female , Mice , Inflammation , Metabolism , Interferon-gamma , Metabolism , Lung , Metabolism , Pathology , Mice, Inbred BALB C , Oligodeoxyribonucleotides , Pharmacology , Phosphorylation , STAT4 Transcription Factor , Metabolism , Silicon Dioxide , Toxicity
17.
Chinese Journal of Applied Physiology ; (6): 101-105, 2013.
Article in Chinese | WPRIM | ID: wpr-358668

ABSTRACT

<p><b>OBJECTIVE</b>To observe the change of apelin and its receptor (APJ) in the lung tissue of rats with pulmonary hypertension induced by monocrotaline and to explore its significance.</p><p><b>METHODS</b>Twenty-five male SD rats were randomly divided into control group (n = 10) and monocrotaline group (n = 15). On the twenty-first day after the rats were intraperitoneally injected 60 mg/kg monocrotaline for monocrotaline group or equal volume vehicle for control group, the mean pulmonary artery pressure was measured by right heart catheterization. Histopathological study of lung tissue was done with hematoxylin-eosin (HE) and Masson's trichrome staining. The concentration of apelin in the plasma was measured by radioimmunoassay. The expressions of apelin/APJ proteins and genes in lung tissue were measured respectively by Western blot and reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The mean pulmonary arterial pressure, right ventricular hypertrophy, pulmonary vascular remodeling index, content of apelin protein in lung tissue of monocrotaline group were higher than those in control group. APJ protein and gene expression in monocrotaline group were significantly lower than those in control group (P < 0.01, P < 0.05), but apelin gene expression in the lung tissue between the two groups had no significant difference.</p><p><b>CONCLUSION</b>Endogenous apelin/APJ dysfunction may play an important role in the development of pulmonary hypertension induced by monocrotaline.</p>


Subject(s)
Animals , Male , Rats , Apelin , Apelin Receptors , Hypertension, Pulmonary , Metabolism , Intercellular Signaling Peptides and Proteins , Metabolism , Lung , Metabolism , Monocrotaline , Rats, Sprague-Dawley , Receptors, G-Protein-Coupled , Metabolism
18.
Asian Pacific Journal of Tropical Medicine ; (12): 407-409, 2013.
Article in English | WPRIM | ID: wpr-820033

ABSTRACT

OBJECTIVE@#To discuss effect of ionizing radiation on transcription of colorectal cancer multidrug resistance (MDR) 1 gene of HCT-8 cells.@*METHODS@#Total RNA was extracted by guanidine thiocyanate one-step method. Northern blot was applied to detect transcription level of MDR1 gene. The expression of P-gp protein was detected by flow cytometry.@*RESULTS@#The expression of MDR1 of normal colorectal cancer HCT-8 cells was low. It was increased by 8.35 times under stimulus with 2 Gy. When treated with low doses in advance, high expressed MDR was decreased significantly under 0.05, 0.1 Gy, which was 69.00%, 62.89% in 2 Gy group and 5.77 times, 5.25 times in sham irradiation group. No obvious difference was detected between (0.2+2) Gy group and 2 Gy group. Compared with sham irradiation group, the percentage of P-gp positive cells after radiation of a high 2 Gy dose was increased significantly (P<0.01). When treated with high radiation dose following low radiation dose (0.05 Gy, 0.1 Gy) in advance, the percentage of P-gp positive cells were also increased significantly. The percentage of P-gp positive cells were increased obviously in 0.2 Gy and 2 Gy groups. Compared with simple high radiation 2 Gy group, the percentage of P-gp positive cells was decreased significantly (P<0.05).@*CONCLUSIONS@#Low radiation dose can reverse multidrug resistance of colorectal cancer cells caused by high radiation dose.


Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Genetics , Cell Line, Tumor , Colorectal Neoplasms , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , RNA, Neoplasm , Genetics , Metabolism , Radiation, Ionizing , Transcription, Genetic , Radiation Effects
19.
Chinese Journal of Gastrointestinal Surgery ; (12): 622-624, 2012.
Article in Chinese | WPRIM | ID: wpr-321563

ABSTRACT

<p><b>OBJECTIVE</b>To study the distribution characteristics and the targeting feature of polyethylene glycol (PEG) modified 5-fluorouracil magnetic albumin microspheres (5-FU-MAMS) and 5-FU-MAMS in major organs of colorectal neoplasm nude mice under magnetic field, and to provide experimental evidence for targeting therapy.</p><p><b>METHODS</b>Eighteen mice were equally divided into PEG-5-FU-MAMS group(n=6), 5-FU-MAMS group(n=6), and 5-FU group(n=6). The colorectal neoplasm was exposed in the magnetic field of 3000 GS for 30 minutes. Three types of 5-FU were injected through the vena caudalis at the dose of 8 mg/kg. Thirty minutes later, the animals were immediately sacrificed after blood draw from the fossa orbitalis. The concentration of 5-FU in different organs including liver, lung, and tumor tissue were determined by the high performance liquid chromatography (HPLC).</p><p><b>RESULTS</b>The 5-FU concentrations in colorectal cancer tissue, liver, lung, and blood were(73.3±3.2), (22.1±2.7), (26.3±2.8), and(1.6±0.6) mg/L in the PEG-5-FU-MAMS group, and were(55.9±5.4), (46.3±8.2), (39.4±5.4), and(1.7±0.4) mg/L in the 5-FU-MAMS group. The 5-FU concentration in colorectal neoplasm was higher in the PEG-5-FU-MAMS group than that in the 5-FU-MAMS group(P<0.01), while the concentration was lower in the liver and the lung than that in the 5-FU-MAMS group(all P<0.01). There were no significant difference of 5-FU concentration in the blood sample(P>0.05).</p><p><b>CONCLUSION</b>Both PEG-5-FU-MAMS and 5-FU-MAMS show significant magnetic targeting to the colorectal neoplasm, and passive target capacity of PEG-5-FU-MAMS to liver and the lung. PEG modification can decrease passive target capacity and the active target capacity can be enhanced, which efficiently reduces the toxicity of chemotherapeutic agents to important organs, and therefore provides a new initiative targeting chemotherapy for cancer.</p>


Subject(s)
Animals , Humans , Mice , Colorectal Neoplasms , Drug Therapy , Metabolism , Fluorouracil , Pharmacokinetics , Magnetics , Mice, Nude , Microspheres , Tissue Distribution , Xenograft Model Antitumor Assays
20.
Journal of Experimental Hematology ; (6): 154-158, 2012.
Article in Chinese | WPRIM | ID: wpr-331000

ABSTRACT

The aim of this study was to explore the effect of mesenchymal stem cell (MSC) conditioned medium (MSC-CM) on proliferation, migration and adhesion of human umbilical vein endothelial cell (CRL1730) and its mechanism. Isolation and purification of MSC were performed with the classic adhering method, the surface markers (CD29, CD90, CD45 and CD34) in MSC were detected by flow cytometry. MSC were treated and cultured for 3 d, the MSC-CM or MSC overexpressing stem cell-derived factor-1 (SDF-1) conditioned medium (Ad-SDF-1-MSC-CM) were collected. Subsequently, CRL1730 cells were treated respectively with 2% FBS-DMEM, 15% FBS-DMEM (control group), MSC-CM or Ad-SDF-1-MSC-CM for 24 h, the proliferation of CRL1730 cells was detected by MTT method. CRL1730 cell migration in vitro was performed by using wound healing system. The adhesion ability of CRL1730 cells was analyzed by microscope. The results indicated that the CRL1730 cells treated with Ad-SDF-1-MSC-CM showed greater proliferative capacity than CRL1730 cells treated with MSC-CM. While adding with AMD3100 5 µmol/L, the blocker of CXCR4, the CRL1730 proliferation mediated by Ad-SDF-1-MSC-CM was significantly reduced. Meanwhile, compared with MSC-CM, Ad-SDF-1-MSC-CM had greater effects for promoting CRL1730 migration and enhancing adhesion ability of CRL1730 cells, these effects were significantly inhibited by AMD3100. It is concluded that MSC-CM promotes the migration and adhesion ability of CRL1730 cells through SDF-1 expressed by MSC.


Subject(s)
Humans , Cell Adhesion , Cell Movement , Cell Proliferation , Cells, Cultured , Culture Media, Conditioned , Human Umbilical Vein Endothelial Cells , Cell Biology , Mesenchymal Stem Cells , Cell Biology
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