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Dependovirus , Terapia Genética , Injeções Espinhais , Atrofias Musculares Espinais da Infância , Humanos , Atrofias Musculares Espinais da Infância/genética , Atrofias Musculares Espinais da Infância/terapia , Terapia Genética/métodos , Lactente , Dependovirus/genética , Masculino , FemininoRESUMO
As widely acknowledged, wastewater treatment plants (WWTPs) stand as significant contributors to the presence of microplastics in surface water. Nonetheless, there exists a notable research gap regarding the extent of potential pollution resulting from the concurrent and uninterrupted discharges originating from multiple WWTPs into small-scale receiving water bodies. This study endeavors to address this knowledge deficit by conducting a thorough investigation into the prevalence of microplastics in surface water. The research encompasses seven distinct locations within the Changzhou section of the Beijing-Hangzhou Grand Canal and the effluent of three WWTPs situated along the tributary. The results indicate differences in the distribution of microplastics in surface waters of mainstream and tributaries. While the microplastic abundance and composition showed little variation along the main stream, the tributaries displayed an overall increasing trend in microplastic abundance from upstream to downstream. Notably, the major contributors to this increase were fragments, fiber particles, and microplastics with particle sizes ranging from 100 to 300 µm. Considering that the primary distinction between the tributaries and the mainstream is the presence of the three WWTPs along the tributaries, the study conducted a correlation analysis between river surface water and effluents from these plants. The results indicated a stronger correlation between the tributaries and the effluents, suggesting that WWTPs are one of the primary factors contributing to the elevated levels of microplastics in the tributaries. Finally, a comparative analysis of microplastic abundance in the Beijing-Hangzhou Grand Canal's Changzhou section and other regions was conducted. The findings revealed that the microplastic pollution level in the Beijing-Hangzhou Grand Canal's Changzhou section is higher than that in most other rivers. Therefore, the issue of microplastic pollution in the Beijing-Hangzhou Grand Canal's Changzhou section warrants our attention, particularly with regard to the effectiveness of microplastic removal by the WWTPs along its course.
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BACKGROUND: The standardised management of neonatal critical care centres can help improve health outcomes of vulnerable newborns. Guidance is required to update evidence related to construction and management of neonatal critical care centres in China. OBJECTIVE: To establish expert consensus on the essential capability lists for neonatal critical care at three levels in China. DESIGN AND SETTING: According to China's administrative divisions, the Chinese guidelines stratifies neonatal critical care into three levels: county level (basic and special care), city level (intensive care) and province level (comprehensive care including neonatal surgery and more subspecialty interventions). A modified Delphi study was conducted. A group of 20 neonatologists from the Chinese Association of Neonatologists rated the importance of capability items on a 5-point Likert scale. RESULTS: At county level, the list consisted of 29 items related to basic and special care, and 3 items were unanimously rated very important by all experts: neonatal resuscitation, endotracheal intubation and continuous positive airway pressure ≥72 hours. At city level, group consensus defined 38 items as essential. Besides the essential items of county level, more items for intensive care were included in city level. At province level, 64 items reached consensus, including neonatal surgery and more advanced subspecialty interventions. The Kendall's W values showed good agreement among experts in both rounds, and an increase from round 1 to round 2. CONCLUSIONS: We developed the capability lists for neonatal critical care at three levels in China. Neonatal resuscitation should be provided by all levels. Interventions for preterm newborns are stratified according to gestational age and birth weight. Congenital abnormalities requiring surgical services need to be managed in high-level centres.
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Cuidados Críticos , Ressuscitação , Humanos , Recém-Nascido , Técnica Delphi , Intubação Intratraqueal , ChinaRESUMO
ObjectiveProtein arginine methyltransferases (PRMTs) play pivotal roles in numerous cellular biological processes. However, the precise regulatory effects of PRMTs on the fate determination of mesenchymal stromal/stem cells (MSCs) remain elusive. Our previous studies have shed light on the regulatory role and molecular mechanism of PRMT5 in MSC osteogenic differentiation. This study aims to clarify the role and corresponding regulatory mechanism of PRMT7 during the adipogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). Methods(1) Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured in a medium that induces adipogenesis. We used qRT-PCR and Western blot to monitor changes in PRMT7 expression during adipogenic differentiation. (2) We created a cell line with PRMT7 knocked down and assessed changes in PRMT7 expression and adipogenic capacity using Oil Red O staining, qRT-PCR and Western blot. (3) We implanted hBMSCs cell lines mixed with a collagen membrane subcutaneously into nude mice and performed Oil Red O staining to observe ectopic lipogenesis in vivo. (4) A cell line overexpressing PRMT7 was generated, and we examined changes in PRMT7 expression using qRT-PCR and Western blot. We also performed Oil Red O staining and quantitative analysis after inducing the cells in lipogenic medium. Additionally, we assessed changes in PPARγ expression. (5) We investigated changes in insulin-like growth factor 1 (IGF-1) expression in both PRMT7 knockdown and overexpressing cell lines using qRT-PCR and Western blot, to understand PRMT7’s regulatory effect on IGF-1 expression. siIGF-1 was transfected into the PRMT7 knockdown cell line to inhibit IGF-1 expression, and knockdown efficiency was confirmed. Then, we induced cells from the control and knockdown groups transfected with siIGF-1 in lipogenic medium and performed Oil Red O staining and quantitative analysis. Finally, we assessed PPARγ expression to explore IGF-1’s involvement in PRMT7’s regulation of adipogenic differentiation in hBMSCs. Results(1) During the adipogenesis process of hBMSCs, the expression level of PRMT7 was significantly reduced (P<0.01). (2) The adipogenic differentiation ability of PRMT7 knockdown group was significantly stronger than that of control group (P<0.001). (3) The ectopic adipogenic differentiation ability of PRMT7 knockdown group was significantly stronger than that of control group. (4) The adipogenic differentiation ability of the PRMT7 overexpression group was significantly weaker than that of the control group (P<0.01). (5) The expression level of IGF-1 increased after PRMT7 knockdown (P<0.000 1). The expression level of IGF-1 decreased after PRMT7 overexpression (P<0.000 1), indicating that PRMT7 regulates the expression of IGF-1. After siIGF-1 transfection, the expression level of IGF-1 in all cell lines decreased significantly (P<0.001). The ability of adipogenic differentiation of knockdown group transfected with siIGF-1 was significantly reduced (P<0.01), indicating that IGF-1 affects the regulation of PRMT7 on adipogenic differentiation of hBMSCs. ConclusionIn this investigation, our findings elucidate the inhibitory role of PRMT7 in the adipogenic differentiation of hBMSCs, as demonstrated through both in vitro cell-level experiments and in vivo subcutaneous transplantation experiments conducted in nude mice. Mechanistic exploration revealed that PRMT7’s regulatory effect on the adipogenic differentiation of hBMSCs operates via modulation of IGF-1 signaling pathway. These collective findings underscore PRMT7 as a potential therapeutic target for fatty metabolic disorders, thereby offering a novel avenue for leveraging PRMT7 and hBMSCs in the therapeutic landscape of relevant diseases.
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Objective To investigate the function of cAMP response element binding protein (CREB) and extracellular signal-regulated kinase (ERK1/2) in osteoclast differentiation mediated by Ca2+/calmodulin-dependent kinase Ⅱ (CaMK Ⅱ)δ,and elucidate the molecular mechanism thereof.Methods CaMK Ⅱδ RNA interference lentivirus vector was constructed and mouse RAW264.7 cells were transfected with the virus to determine the interference efficiency.After virus transfection,RAW264.7 cells were treated with 50ng/ml receptor activator of nuclear factor κB ligand (RANKL) and the phosphorylation levels of CREB and ERK1/2 were detected at different time points.The cells were also treated with PD98059,an ERK1/2 inhibitor,to determine the effect of ERK1/2 signal blocking on the expression of nuclear factor activated T-cells cytoplasmic 1 (NFATc1) and osteoclast differentiation.Results Interference efficiency of recombinant CaMK Ⅱδ virus vector was 77.2% at mRNA level and 70.2% at protein level.CaMK Ⅱδ RNA interference significantly suppressed phosphorylation of CREB and ERK1/2,and the levels ofp-CREB and p-ERK1/2 were down-regulated by 21%-55% and 55%-64%,respectively.ERK1/2 inhibitor significantly down-regulated the protein expression of NFATc1,and the number of osteoclast,the number and size of bone resorption lacunae decreased by 39.3%,50.0% and 52.3%,respectively.Conclusion CaMK Ⅱδ RNA interference may significantly suppress the phosphorylation of CREB and ERK1/2,and CREB and ERK1/2 have mediated the CaMK Ⅱδ-induced osteoclast differentiation.
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The nonstructural protein 1 (NS1) of influenza A virus, which is absent from the viral particle, but highly expressed in infected cells, strongly antagonizes the interferon (IFN)-mediated antiviral response. We engineered an NS1-expressing 293 (293-NS1) cell line with no response to IFN stimulation. Compared with the parental 293 cells, the IFN-nonresponsive 293-NS1 cells improved the growth capacity of various viruses, but the introduction of NS1 barely enhanced the propagation of Tahyna virus, a negative-strand RNA virus. In particular, fastidious enteric adenovirus that replicates poorly in 293 cells may grow more efficiently in 293-NS1 cells; thus, IFN-nonresponsive 293-NS1 cells might be of great value in diagnostic laboratories for the cultivation and isolation of human enteric adenoviruses.
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Humanos , Linhagem Celular , Regulação da Expressão Gênica , Células HEK293 , Vírus da Influenza A , Fisiologia , Proteínas não Estruturais Virais , Genética , Metabolismo , Cultura de Vírus , Métodos , Replicação Viral , FisiologiaRESUMO
OBJECTIVE: To develop a microfluidic mammalian cell culture device with human breast cancer cells co-cultured with extracellular matrix (ECM), reconstitute the 3-dimension human body microenvironment and preliminarily conduct drug screening on the breast cancer chip. METHODS: 3D Printing technology was utilized to build a two-layer microchip with cell culture reservoirs. The breast cancer cells (MCF7) were cultured in the matrix gel structure which mimiced the 3-dimension human body microenvironment. The cell toxicity of paclitaxel (PTX) on MCF7 cells was preliminarily observed using this microfluidic chip. With the use of AO/EB immunofluorescent staining and laser confocal microscopy, the cell death ratio induced by PTX was determined and compared with that determined by 2-dimension drug screening methods. RESULTS: MCF7 cells cultured on the chip successfully formed a 3D cavity structure in the matrix after 6-day dynamic incubation. The flow rate was 10 μL · h-1. After 24 h dynamic culture, the culture medium was changed to culture solution containing 0.02 μmol · mL-1 PTX, and the incubation was continued for 24 h. Obvious cell death was detected under laser confocal microscopy. CONCLUSION: The microfluidic chip developed in this study can successfully culture breast cancer cells in 3-dimension structure and perform drug screening, which lays a foundation for actualization of "human-on-a-chip".
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To investigate the morphogenetic process of human adenovirus type 41 (HAdV-41), 293TE cells were infected with purified wild-type HAdV-41, and ultrathin sections of infected cells were prepared and observed under a transmission electron microscope. Results showed that HAdV-41 entered host cells mainly through three ways: non-clathrin-coated pit, clathrin-coated pit, and direct penetration of plasma membrane. In addition, cell microvilli might help HAdV-41 enter cells. After entering into cells, HAdV-41 virus particles could be found in vacuoles or lysosomes or be in a free state in cytoplasm. Only free virus particles could be found near nuclear pores (NP), suggesting that the virus needed to escape from lysosomes for effective infection and viral nucleoprotein entered the nucleus through NP. Progeny viruses were as-sembled in the nucleus. Three types of inclusion bodies, which were termed as fibrillous inclusion body, condense inclusion body, and stripped condense inclusion body, were involved in HAdV-41 morphogenesis. In the late phase of viral replication, the membrane integrity of the infected cells was lost and viral particles were released extracellularly. This study reveals the partial process of HAdV-41 morphogenesis and provides more biological information on HAdV-41.
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Humanos , Infecções por Adenovirus Humanos , Virologia , Adenovírus Humanos , Genética , Fisiologia , Membrana Celular , Virologia , Núcleo Celular , Virologia , Liberação de Vírus , Replicação ViralRESUMO
To investigate the components of fibrillous inclusion body (FIB), which was formed in packaging cells during the replication of human adenovirus type 41 (Ad41), Ad41 long fiber knob (LFK) and short fiber knob (SFK) proteins were expressed in prokaryote respectively and then used to immunize BALI mice for preparation of anti-LFK serum and anti-SFK sera. The activity and specificity of anti-LFK and an ti-SFK sera were confirmed with Western blot, indirect immunofluorescence assay (IFA) and immunonegative staining, suggesting these sera could be applied in immuno-colloidal gold labelling electron microscopy (EM). 293TE cells were infected with wild-type Ad41. Ultrathin sections of infected cells were made, and labelled with immuno-colloidal gold technique using anti-Ad41 sera, anti-LFK sera, anti-SFK sera, or anti-fiber monoclonal antibody 4D2, respectively. The labelled sections were observed under EM, and the results demonstrated that both Ad41 long fiber protein and short fiber protein were included FIB.
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Animais , Feminino , Humanos , Camundongos , Infecções por Adenovirus Humanos , Virologia , Adenovírus Humanos , Genética , Metabolismo , Corpos de Inclusão Viral , Camundongos Endogâmicos BALB C , Microscopia ImunoeletrônicaRESUMO
<p><b>OBJECTIVE</b>To investigate the genetic stability of non-replicating recombinant adenovirus which used Ad41 as vector and could express VP6 gene of group A rotavirus during continous passage, in order to develop the vaccine of rotavirus.</p><p><b>METHODS</b>The recombinant adenovirus rvAd41-VP6 (o) was prepared by our laboratory early, it then was continuously propagated on 293TE7 cells for 14 passages. After that samples of the infected cells were collected at every 2 passages for the detection of the integration of the VP6 gene by PCR, and the expression of the target protein was detected by Western Blot analysis.</p><p><b>RESULTS</b>Analysis by PCR revealed that, there was stable integration of specific VP6 gene in the rvAd41-VP6 (o), Western Blot analysis confirmed that rvAd41-VP6 (o) could stably expressed the group-specific antigen structural protein VP6 (o), and it had preferable genetic stability.</p><p><b>CONCLUSION</b>The recombinant adenovirus rvAd41-VP6 (o) which could stably express the VP6 (o) gene had favorable biological property in vitro, and it has provided a basis for further research of animal immunization.</p>
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Humanos , Adenoviridae , Genética , Metabolismo , Antígenos Virais , Genética , Metabolismo , Proteínas do Capsídeo , Genética , Metabolismo , Linhagem Celular , Expressão Gênica , Vetores Genéticos , Genética , Metabolismo , Rotavirus , Genética , Metabolismo , Infecções por Rotavirus , VirologiaRESUMO
In order to optimize the ~(18)O labeling method, two key aspects, peptide dispersion and trypsin deac tivation were discussed o The addition of Rapigest SF in H_2~('8)O and microwave heating enhanced labeling efficiency of α-casein digested peptides(~(18)O/~(16)O) ratio >99%).Chemical modification with tris(2-carboxyeth yl) phosphine (TCEP) and iodoacetamide (IAA) resulted in trypsin deactivated completely.No significant back-exchange from ~(18)O to ~(16)O was observed after labeling in 6 days.The experiment result with peptide mixture from showed that the improved method could be effectively used to label protein and peptide.
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<p><b>OBJECTIVE</b>To establish a TaqMan-based Real-time PCR assay with internal control for the detection of parvovirus B19 DNA in human serum.</p><p><b>METHODS</b>Two DNA fragments in length of 113 bp each were artificially synthesized, cloned into T vector and used as standard DNA or internal control, respectively. One pair of primers and two probes were included in the Real-time PCR. The probes were labeled with different fluoresceins and could bind B19 DNA or internal control, respectively. Precision and specificity of the method were evaluated. Specimens of human serum were examined by this assay to find B19 DNA.</p><p><b>RESULTS</b>The standard curve was constructed using the quantified standard B19 DNA. The Real-time PCR method was established. It was stable according to precision evaluation by the intra- and inter-assay and specific without any evident cross-reaction with human hepatitis B virus (HBV). Among 160 samples of human serum, B19 DNA was detected in 2 with a concentration of 2.1 x 10(5) Geq/ml and 3.6 x 10(3) Geq/ml, respectively.</p><p><b>CONCLUSION</b>The Real-time PCR for B19 DNA detection was developed successfully, in which the internal control was helpful to exclude false-negative results.</p>
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Humanos , DNA Viral , Sangue , Parvovirus B19 Humano , Genética , Reação em Cadeia da Polimerase , MétodosRESUMO
Biological mass spectrometry has been developed for the largE-scale protein identification. The successful identification of protein in proteomic study is based on an effective match of MS data to the sequence in database. Because of the diversity and heterogeneity of protein modification, the experimental data obtained by mass spectrometry does not match the theoretical value sometimes, which makes about 90 percent or more of the tandem mass spectra not be effectively identified. This has become one of the most important technique issues to be resolved in current proteome research. The N-terminal cyclization of peptides, as one of a variety of modification introduced in sample preparation, has been preliminarily studied in this work. The result showed that N-terminal cyclization occurred at the most of the glutamine(Q) or carbamoylmethyl-cysteine(CAM_C) residues and the reaction is often incomplete or partial, both types of peptides could often exist in its respective state at the same time, and the behavior of modified peptides in revered phase chromatography is also changed. The success rate of protein identification could be obviously improved if adding the N-terminal cyclization modification in the database searching. These results will be very helpful in the mass spectrometric data analysis of proteomic study.
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Adenovirus type 40 and 41 (Ad40, Ad41), which belong to human adenovirus subgroup F, are called fastidious adenoviruses due to their property of poor growth in cultured cell lines in vitro The effect of expression of exogenous E1B55K in Hep2 on Ad41 replication in this cell line was investigated. E1B55K gene was amplified by PCR with DNA extracted from Ad41-positive feces supernatant as template. Eukaryotic expression plasmid (pcDNA3) carrying E1B55K was constructed, purified, and transferred into Hep2 cell. Expression of E1B55K in G418-resistant clones was assayed by RT-PCR, and one clone named as Hep2-E1B4#4 could produce more Ad41 progenies when compared with other clones by the method of inducing complete cytopathic effect (CPE) in 293 cells. Infection of equivalent Ad41 caused more significant cytopathic effect (CPE) in Hep2-E1B#4 than that in the control cells of Hep2 or Hep2-DNA3, also suggesting enhanced viral replication in Hep2-E1B#4. The titer of Ad41 was further determined by method of immunocytochemical staining, and semi-quantity PCR was employed to compare the copy number of Ad41 genome DNA. The results showed that the yield of Ad41 in Hep2-E1B#4 was more than 9 times of that in control cells when equal amount of seed viruses were incubated, and the copy number of Ad41 genome increased 4 times in the raw extract from the infected Hep2-E1B#4 when compared with that from control cells. In conclusion, E1B55K gene transfer improved the ability of Hep2 in packaging Ad41, and the Hep2-E1B#4 cell line, which expressed E1B55K constitutively, would be helpful in isolation, cultivation and amplification of Ad41.
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Humanos , Proteínas E1B de Adenovirus , Genética , Metabolismo , Adenovírus Humanos , Genética , Linhagem Celular Tumoral , Expressão Gênica , Imuno-Histoquímica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Replicação Viral , GenéticaRESUMO
Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease of transformed hematopoietic progenitor cells. It is now clear that the chimeric bcr/abl P210(bcr/abl) fusion protein, which is generated by the reciprocal translocation t (9; 22), inhibits apoptosis and increase proliferation. P210(bcr/abl) plays a central role in the pathophysiology of CML. The purpose of this study was to construct a cell line model that bcr/abl expression can be regulated by Tet-off inducing-expression-system. The full-length b3a2 bcr/abl cDNA was subcloned into the pTRE2hyg expression vector to construct the pT2-P210 plasmid. 293 cells were firstly transfected with Tet-off plasmid and the clone that the Tet-off system can work effectively after transfected with pTRE2hyg-LUC was selected by luciferase activity assay. The pT2-P210 plasmid was then transfected into the selected clone and cells were then selected for hygromycin B and G418 resistance. The results showed that individual subclones expressing bcr/abl after withdrawing doxycycline were 293pT2-P210 cell line. In conclusion, selected 293pT2-P210 cells are cells that bcr/abl expression can be regulated by Tet-off inducing-expression-system. They are suitable thoroughly to study the function of bcr/abl fusion gene and its signal regulation mechanism.
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Humanos , Sequência de Bases , Linhagem Celular Transformada , Biologia Celular , Fisiologia , Cromossomos Humanos Par 22 , Genética , Cromossomos Humanos Par 9 , Genética , Proteínas de Fusão bcr-abl , Genética , Regulação Neoplásica da Expressão Gênica , Genes abl , Genética , Leucemia Mielogênica Crônica BCR-ABL Positiva , Genética , Metabolismo , Patologia , Modelos Genéticos , Dados de Sequência Molecular , Proteínas Proto-Oncogênicas c-bcr , Genética , Transfecção , Translocação Genética , Células Tumorais CultivadasRESUMO
The aim of this research was to understand the influence of rhG-CSF on the sphingosine kinase (SphK) activity of monocytes. The peripheral blood monocytes were collected from 6 peripheral blood progenitor cell donors on the fifth day of mobilization with rhG-CSF and from 5 blood donors' buffy coats. The mRNA expressions of monocyte G-CSF receptor and SphK were tested with RT-PCR. The changes of SphK activity of monocytes were assayed after being treated with rhG-CSF. The results showed that the two kinds monocytes collected from both blood donors and peripheral blood progenitor cell donors mobilized with rhG-CSF expressed mRNA of G-CSF receptor and SphK. The SphK activity of monocytes collected from blood donors was not changed significantly after being treated with rhG-CSF (P > 0.05). The SphK activity of monocytes collected from peripheral blood progenitor cell donors transiently increased by (39.6 - 87.2)% after being treated by means of rhG-CSF (P < 0.05) without obviously dose-dependent effect. It is concluded that the SphK activity of monocytes collected from peripheral blood progenitor cell donors can be activated by rhG-CSF.
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Humanos , Fator Estimulador de Colônias de Granulócitos , Farmacologia , Mobilização de Células-Tronco Hematopoéticas , Monócitos , Biologia Celular , Fosfotransferases (Aceptor do Grupo Álcool) , Metabolismo , Receptores de Fator Estimulador de Colônias de Granulócitos , Genética , Proteínas RecombinantesRESUMO
<p><b>OBJECTIVE</b>To probe into the mechanisms of thread implantation at Zusanli(ST 36) and Chinese herbs in treatment of chronic renal failure(CRF).</p><p><b>METHODS</b>CRF rat model was made by Platt subtotal nephrectomy. They were divided into 5 groups, sham operation group, model group, Chinese herbs group, thread implantation group and thread implantation plus Chinese herbs group. After treatment of 8 weeks, serum parathyroid hormone (PHT), transforming growth factor beta1 (TGF-beta1) expression in residual renal cells, malondialdehyde(MDA) content in the residual renal tissue, serum urea nitrogen(BUN) and creatinine(Scr), protein in urine and pathological changes were investigated.</p><p><b>RESULTS</b>The above indexes after treatment by thread implantation at acupoint, Chinese herbs, and acupoint thread implantation plus Chinese herbs showed begin reversion, especially, the most obviously improvement in the acupoint thread implantation plus Chinese herbs treatment group.</p><p><b>CONCLUSION</b>The mechanism of acupoint thread implantation and Chinese herbs in improvement of CRF is related with decrease of PTH, inhibition of TGF-beta1 expression, decrease of MDA content and resisting lesion of renal fibrosis.</p>
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Animais , Masculino , Ratos , Pontos de Acupuntura , Medicamentos de Ervas Chinesas , Farmacologia , Falência Renal Crônica , Sangue , Terapêutica , Hormônio Paratireóideo , Sangue , Ratos Wistar , Fator de Crescimento Transformador beta , Sangue , Fator de Crescimento Transformador beta1RESUMO
The study was to understand the impact on the proliferation and cytotoxicity of donor's T cells during mobilization with rhG-CSF. The peripheral blood mononuclear cells (PBMNC) were collected from 15 donors before mobilization and on fifth day of mobilization with rhG-CSF. After the PBMNC were activated with 500 ng/ml of CD3 monoclonal antibody and 500 microg/ml of rhIL-2 for 96 hours, the activated T cells were collected for testing proliferation, cytotoxicity, Fas expression, perforin and Fas ligand (FasL) mRNA expression, the IFN-gamma concentration in the culture medium of the activated T cells was determined by radioimmunoassay. The results showed that the proliferation activity of T lymphocytes and the cytotoxicity of T cells activated with CD3 monoclonal antibody and rhIL-2 were reduced markedly after mobilization with rhG-CSF (P < 0.05). The Fas molecule expression in the activated T cells was very high both before and after mobilization with rhG-CSF (P > 0.10). The activated T cells expressed perforin mRNA and didn't express FasL mRNA both before and after mobilization with rhG-CSF. The concentration of IFN-gamma in the culture medium of the activated T cells decreased significantly after mobilization with rhG-CSF (P < 0.01). It is concluded that activity of proliferation and cytotoxicity of donor's T cells is impaired after mobilization with rhG-CSF.
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Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proliferação de Células , Células Cultivadas , Proteína Ligante Fas , Genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Farmacologia , Mobilização de Células-Tronco Hematopoéticas , Métodos , RNA Mensageiro , Genética , Proteínas Recombinantes , Linfócitos T , Biologia Celular , Linfócitos T Citotóxicos , Alergia e Imunologia , Receptor fas , GenéticaRESUMO
<p><b>AIM</b>To elucidate the effect of sphingosine kinase (SPK) on the hepatocyte growth factor (HGF)-induced migration of endothelial cells.</p><p><b>METHODS</b>We constructed recombinant adenoviral vectors, which contain SPK gene and its mutant respectively. These adenoviral vectors were packaged and amplified in 293 cells. And intracellular SPK activity was assayed via measurement of [32]P radioisotope labeled S1P; the effect of SPK activation on HGF-induced migration of endothelial cell was observed by Transwell technique.</p><p><b>RESULTS</b>Adenoviral mediated expression of SPK gene increased in ECV 304 cells intracellular SPK activity, which in turn enhanced the HGF-induced migration. Whereas these activities were blocked by the dominant negative SPK gene.</p><p><b>CONCLUSION</b>These findings show that SPK activation plays important roles in the regulation of HGF-induced migration of endothelial cells.</p>
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Humanos , Adenoviridae , Metabolismo , Linhagem Celular , Movimento Celular , Células Endoteliais , Biologia Celular , Fator de Crescimento de Hepatócito , Farmacologia , Fosfotransferases (Aceptor do Grupo Álcool) , Genética , Metabolismo , Transdução de SinaisRESUMO
<p><b>OBJECTIVE</b>To study the mechanism of auricular acupuncture for improvement of learning and memory disorders in the rat of vascular dementia (VD).</p><p><b>METHODS</b>The vascular dementia rat model was made by 4-vessel occlusion method. Four groups, a sham operation group, a normal control group, a model group and an auricular acupuncture group were set up. After acupuncture was given at auricular points, Brain and Kidney. Immunohistochemical analysis, behavioural observation and computer image analysis were made.</p><p><b>RESULTS</b>Auricular acupuncture could decrease significantly the beta-amyloid protein (A beta) immunoreactivive neurons and increase its average optical density in the parietal cortex of the VD rats (P < 0.05).</p><p><b>CONCLUSION</b>Auricular acupuncture can reduce or inhibit the over-production of Abeta in the brain, so as to improve the learning and memory capacity of the VD model rat.</p>