AIM: To observe the effect of microRNA-16 (miR-16) on the megakaryocytic differentiation of K562 cells, and to explore the potential mechanism.METHODS:miR-16 was over-expressed or silenced by transfection with miR-16 mimics or inhibitor in K562 cells.The level of miR-16 was detected by real-time PCR.The expression of CD41, CD42b and CD61, as megakaryocytic differentiation markers, was detected by flow cytometry.The effect of miR-16 on the expression of myeloblastosis oncogene ( MYB) was measured by Western blotting, and flow cytometry was performed to confirm whether the effect of miR-16 on expression of CD41, CD42b and CD61 was mediated by MYB.RESULTS:Transfection with miR-16 mimics dramatically elevated the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells.Transfection with miR-16 inhibitor decreased the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells (P<0.05).The expression of MYB was regulated by miR-16, and MYB silencing reversed the regulation of CD41, CD42b and CD61 induced by miR-16.CONCLUSION:miR-16 regulates the megakaryocytic dif-ferentiation of K562 cells by targeting MYB.

Amino Acids ; urine ; Animals ; Biogenic Monoamines ; metabolism ; urine ; Male ; Microwaves ; Rats ; Rats, Wistar

Objective To provide references for enriching and innovating patterns of TCM syndrome differentiation by exploring the relationship between pathological changes and TCM syndrome differentiation of delayed graft function (DGF) after renal transplantation. Methods Totally 57 patients with DGF after renal transplantation who were diagnosed by pathological biopsy were also diagnosed by TCM doctors to obtain TCM syndromes. And the pathological diagnoses were compared with the distribution of TCM syndromes to discuss the relationship between them. Results All the patients were classified into four TCM syndromes:spleen-kidney qi deficiency, liver-kidney yin deficiency, qi-stagnation and blood stasis, and deficiency of both qi and yin. The results of pathological biopsy were as follows:acute tubular necrosis, calcineurin inhibitor toxicity and acute rejection. Quantitative corresponding relation was found between the pathological diagnoses and the TCM syndromes. Conclusion There are inner links between pathological diagnosis and TCM syndromes of DGF, which can be references for diagnosis and treatment of TCM syndrome differentiation.

Objective To evaluate the characteristics of abdominal aorta wall motion in different stages of rats atherosclerosis with velocity vector imaging (VVI) technique. Methods Twenty-four healthy SD rats were on high-fat feeding after one week ordinary diet. Abdominal aortic intima-media thickness (IMT), end-systolic blood vessel diameter (Ds), peak systolic velocity (Vs), resistance index (RI), pulsatility index (PI) were measured before and at the end of 8th and 12nd week. Artery wall peak velocity (V_(max)), maximum tangential strain (S_(max)) and the maximum tangential strain rate (SR_(max)) were caculated with VVI. Results Abdominal aortic intima was rough and a small amount of foam cells were found under the light microscope at the end of 8 weeks of high-fat feeding. The values of Smax and SRmax measured at the end of 8th week of high-fat feeding decreased significantly than those of before high-fat feeding (P<0.05). At the end of 12nd week, abdominal aortic intimal was thicker and atherosclerotic plaque appeared somewhere. There were significant differences in artery IMT, Ds, Vs, RI, PI between before and the end of 2nd week of high-fat feeding (P<0.05);the values of V_(max), S_(max), SR_(max) decreased significantly than those of before and at the end of 8th week of high-fat feeding (P<0.05). Conclusion VVI can quantitatively evaluate the vessel wall elasticity in different stage of arteriosclerosis rats.

Objective To explore the impact of dexamethasone on inflammatory response of thyrocytes.Methods Primary thyrocytes were extracted from thyroid tissue of patients with Graves' disease.The cells were stimulated with interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α),and cultured in dexamethasone.Thyrocytes were divided into 4 groups:control group,dexamethasone group,TNF-α + IFN-γ group,and dexamethasone+TNF-α+IFN-γ group.Interferon-γ-induced protein 10 (CXCL10) and CCL2 in supernatant of cell cultured in 4 groups were detected by enzyme-linked immunosorbent assay.Cell protein in 4 groups was extracted and GSK-3β,P50,and P100 protein were detected by Western blotting.Results MTT assay demonstrated that 10-5 mmol/L concentration of dexamethasone was optimal for cell culture.The CXCL10 level in TNF-α+IFN-γ group was higher than that in the control group and dexamethasone group (P＜0.01),but no difference was found between dexamethasone+TNF-α+IFN-γgroup and TNF-α+IFN-γgroup(P＞0.05).The CCL2 level in TNF-α+IFN-γ group was higher than that in control group and dexamethasone group(P＜0.01).There was a significant lowering of CCL2 level in dexamethasone + TNF-α + IFN-γgroup compared with TNF-α + IFN-γ group (P ＜ 0.05).The expression of GSK-3β and P100 protein was increased in TNF-α + IFN-γgroup compared with control group.The expression of GSK-3β and P100 protein was lower in dexamethasone+TNF-α + IFN-γ group than that in TNF-α + IFN-γ group.Conclusion TNF-α + IFN-γ could stimulate the secretion of CXCL10 and CCL2 in thyrocytes and thus activate the inflammatory response.Dexamethasone could reduce CCL2 secretion.Dexamethasone had little effect on CXCL10.Dexamethasone could reduce GSK-3β and P100 expressions,and inhibit the activation of NF-κB signaling pathway and thus the inflammatory response.

The study reports the detection of neuroprotective effect of 10 kinds of coumarin derivatives and explores their possible mechanism. MTT method was used to screen the neuroprotective effect of 10 coumarin derivatives on neurotoxic agents (Aβ25-35 and rotenone) or OGD (oxygen-glucose deprivation). A compound with better protective effect was obtained. Then the effect of this compound on neurotoxic agents on PC12 was detected by the morphological observation. Furthermore, the effect of compound 3 on microglia with lipopolysaccharide (LPS) induced inflammation was detected. And the inflammatory factor was tested. Finally, direct free radical scavenging ability was detected. Compound 3 was found to be the best compound through three neurons toxic models. Not only compound 3 ameliorated cell viability reduced by three neurons toxic models, but also significantly inhibited the production of inflammatory factor (TNF-α and IL-1β). And its free radical scavenging ability is very good, especially the effect on superoxide anion, which is comparable with vitamin C. The significant scavenging effect of compound 3 on superoxide anion might be the mechanism of the neuroprotection. Compound 3 as a potential neural cell protective agent merits further investigation.
Animals ; Coumarins ; chemistry ; Free Radical Scavengers ; chemistry ; Inflammation ; Microglia ; drug effects ; Neurons ; drug effects ; Neuroprotective Agents ; chemistry ; PC12 Cells ; Rats

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; DNA, Neoplasm ; genetics ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Infant ; Lymphoma, B-Cell ; diagnosis ; genetics ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; genetics ; Male ; Middle Aged ; Paraffin Embedding ; Polymerase Chain Reaction ; Young Adult

Using brain microdialysis and LC-MS/MS to detect acetylcholine in rat brain to investigate the effects of ligustrazine. A liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of acetylcholine in rat brain dialysate sampling by microdialysis. The results indicated that ligustrazine administration by subcutaneous injection significantly increased Ach release in rat medial prefrontal cortex and nucleus accumbens in a dose-related manner. The drug' s effect on Ach release in rat brain could be directly detected by microdialysis combined with HPLC-MS/MS and this method is selective and sensitive.
Acetylcholine ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Ligusticum ; chemistry ; Male ; Microdialysis ; Nucleus Accumbens ; metabolism ; Plants, Medicinal ; chemistry ; Prefrontal Cortex ; metabolism ; Pyrazines ; administration & dosage ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

An increasing number of monopartite begomoviruses are being identified that a satellite molecule (DNAbeta) is required to induce typical symptoms in host plants. DNAbeta encodes a single gene (termed betaC1) encoded in the complementary-sense. We have produced transgenic Nicotiana benthamiana and N. tabacum plants expressing the betaC1 gene of a DNAbeta associated with Tomato yellow leaf curl China virus (TYLCCNV), under the control of the Cauliflower mosaic virus 35S promoter. Transgenic plants expressing betaC1 showed severe developmental abnormalities in both species. Microscopic analysis of sections of both transgenic and non-transgenic N. tabacum leaves showed abnormal outgrowths of transgenic N. tabacum to be due to disorganized cell division (hyperplasia) of spongy and palisade parenchyma. Immuno-gold labeling of sections with a polyclonal antibody against the betaC1 protein showed that the betaC1 protein accumulated in the nuclei of cells. The possible biological function of the betaC1 protein was discussed.
Cell Division ; physiology ; Cell Nucleus ; genetics ; metabolism ; ultrastructure ; Cells, Cultured ; DNA, Viral ; genetics ; Geminiviridae ; genetics ; Plant Diseases ; genetics ; virology ; Plant Leaves ; cytology ; genetics ; growth & development ; metabolism ; Plants, Genetically Modified ; growth & development ; metabolism ; Recombinant Proteins ; metabolism ; Tobacco ; cytology ; growth & development ; metabolism ; ultrastructure ; Viral Proteins ; genetics ; metabolism

Objective To explore the non-bacteria etiology of pneumonia in children under 12 years old in southwest Shanghai,and investigate clinical characteristic of pneumonia caused by different pathogens.Methods The serum of 187 children with pneumonia from July 2002 to December 2004 in hospital were investigated for respiratory syncytial virus(RSV),adenovirus 3 influenza viruses(IFV) A and B,parainfluenza viruses(PIV)type 1,2,3,4 and coxsackievirus A 1.7,echovirus 7 by employing the indirect immunofluorescence assay(IFA)for the identification of nearly 8 different viruses,and 3 different enteroviruses.Based on the principle that sensitized particles were agglutinated by the pressence of antibodies to mycoplasma pneumonia in human serum.Results Examination for 8 kinds of conventional respiratory virus infected,a total of 90 positive results in 187 cases(48.13%),Firstly was RSV(19.79%),(secondly) was IFV B(16.58%).Out of these 1084 children,154 cases(14.21%)showed positive in anti-mycoplasma pneumonia.Conclusions RSV is still the leading cause of pneumonia in children during winter and spring in southwest in Shanghai.Mycoplasma pneumonia is having been the major pathogens of the school-aged children with pneumonia.