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1.
Zhongguo Zhong Yao Za Zhi ; (24): 1769-1782, 2021.
Artigo em Chinês | WPRIM | ID: wpr-879091

RESUMO

NAC(NAM/ATAF/CUC) protein plays an important role in plant growth and development, secondary cell wall formation and stress response. In this study, based on the sequencing data of Angelica dahurica, the NAC family was systematically analyzed using bioinformatics methods and its expression pattern was analyzed. Studies showed that 75 candidate genes had been selected from the NAC transcription factor family of A. dahurica, with the protein size of 148-641, all of which were unstable hydrophilic proteins. Most NAC proteins were localized in the nucleus, and had complete NAC domain. Phylogenetic analysis of NAC family proteins of A.dahurica and Arabidopsis thaliana showed that among the 17 subfamilies, NAC members were unevenly distributed in each subfamily, indicating that the evolution of species is developing in multiple directions. Among them, ANAC063 subfamily contained no NAC sequence of A. dahurica, which might be due to the functional evolution of the species. Analysis of protein transmembrane structure and signal peptide showed that NAC transcription factor could carry out transmembrane transportation, but its signal peptide function had not been found. Expression analysis showed that most transcription factors responded to abiotic stress and hormones to varying degrees, and the effects of hormones were obvious, especially ABA and IAA. In different organs of A. dahurica, most members of the NAC family had higher expression in root phloem, followed by root xylem. This study lays a foundation for further research on the function of A. dahurica NAC transcription factor and for solving the biological problems of A. dahurica.


Assuntos
Angelica , Biologia Computacional , Regulação da Expressão Gênica de Plantas , Filogenia , Proteínas de Plantas/metabolismo , Estresse Fisiológico , Fatores de Transcrição/metabolismo
2.
Artigo em Chinês | WPRIM | ID: wpr-872859

RESUMO

Professor XUE Bo-shou,a master of traditional Chinese medicine(TCM),believes that coronavirus disease-2019 (COVID-19) belongs to the category of "cold epidemic" or "cold-dampness epidemic" in TCM theory. Cold-dampness obstructing the lungs is the key pathogenesis in the early stage of the disease. Exogenous pathogenic factors of cold and dampness are converted into heat in the interior. Over time,the pathogenesis is the disturbance in ascending and descending of Qi movement,loss of vital Qi, deficiency of both Qi and Yin,and even the occurrence of inner blocking causing Yang collapse. Professor XUE believes that the clinical syndrome differentiation and treatment shall be based on six meridians, and the efficacy could be improved by reference to classical prescriptions of plague in the past dynasties. To treat the disease in the early stage, the prescriptions emphasize on diffusing the lungs,inducing resuscitation and expelling pathogenic factors. He advocates the proper use of Ephedrae Herba, which can induce resuscitation of the lungs,diffuse the lungs and expel pathogenic factors,dissipate the interstitial depression and pathogenic factors in the lung interstitium. Therefore,it is considered that Ephedrae Herba is an effective medicine for diffusing the lungs,expelling pathogenic factors,inducing diuresis,eliminating dampness,promoting blood circulation and removing stasis due to cold coagulation. At the same time,the weakness of vital Qi is the root cause of the disease. The prosperity and decline of vital Qi plays a decisive role in the development, prognosis and efficacy of the disease. Professor XUE attaches importance to overall treatment,and emphasize the basis of pathogenesis, vital Qi, and stomach Qi. We shall expel pathogenic factors of disease,while protecting the vital Qi and stomach Qi. This article summarizes Professor XUE's diagnosis and treatment of COVID-19, enumerates and analyzes professor XUE's consultation cases of COVID-19 in designated hospitals in Linyi, Xinjiang and Beijing, in order to provide more ideas for the anti-epidemic effect of TCM.

3.
Journal of Medical Postgraduates ; (12): 739-744, 2018.
Artigo em Chinês | WPRIM | ID: wpr-818055

RESUMO

Objective Few studies have paid attention to time-zero renal biopsy in living kidney transplantation so far. This article aimed to investigate the risk factors of latent pathologic changes in living donors by time-zero renal biopsy (TO-RBx) and the predictive value in the allograft function of recipients early after living kidney transplantation.Methods We retrospectively analysed the clinical data of 89 renal transplant recipients and living donors who received TO-RBx at Nanjing General Hospital from January 2008 to December 2016. According to the 2007 Banff criteria, the common pathologic changes in living donors such as latent glomeruloscerosis (GS), tubular atrophy (CT), interstitial fibrosis (CI), arteriolar hyaline thickening (AH) and vascular fibrous intimal thickening (CV) were scored. To analyze the influencing factors for different pathological changes and evaluate its predictive value in the allograft function of recipients in 1, 3, 6 months after living renal transplantation.Results Of all the TO-RBx specimens, 23 cases (25.84%) with GS (21 were mild change, 1 was moderate change and 1 was severe change), 33 cases (37.08%) with CT/CI changes (30 were mild change and 3 were moderate change) and 37 cases (41.57%) with AH/CV changes (36 were mild change and 1 was moderate change). GS was related to the donor age (P=0.042); CT/CI changes were related to donor age, gender and systolic pressure (P=0.019;0.006;0.01); arterial changes were related to donor gender and blood triglyceride level (P=0.029;0.049). Within 3 and 6 months after living donor renal transplantation, the eGFR of renal transplant recipients with GS lesions \[(65.96±17.17), (69.52±19.1)mL/min·1.73m2\] were significantly lower than the groups without lesions \[(76.91±18.98), (79.52±18.91)mL/min·1.73m2\] (P<0.05).Conclusion Time-zero renal biopsy has significance in terms of predicting the allograft function in 6 months after transplantation. It can guide the formulation and adjustment of postoperative immunosuppressive regimens for recipients. Besides, it can also detect the latent pathologic changes in living donors and is one of the important evidence for establishing a personalized follow-up plan for donors after surgery. This method is practical in clinical.

4.
Journal of Experimental Hematology ; (6): 1112-1119, 2015.
Artigo em Chinês | WPRIM | ID: wpr-274083

RESUMO

<p><b>OBJECTIVE</b>To investigate the feasibility of umbilical cord blood plasma (UCP) as a replacement for fetal bovine serum (FBS) for culturing mesenchymal stem cells (MSC) derived from umbilical cord, and to observe the supporting effects of these cells (served as a feeder layer) on ex vivo expanding of human umbilical cord blood CD34(+) cells.</p><p><b>METHODS</b>Umbilical cord blood (UCB) units were suitable if the Guangzhou cord blood bank donor selection criteria strictly were fulfilled. UCP were ready to use after the collection from the plasma depletion/reduction during the processing and pooling of suitable UCB units (at least 30 units were screened for pathogens and microorganisms, and qualified). Umbilical cord mesenchymal stem cells (UCMSC) were harvested from the umbilical cord tissue of health full-term newborns after delivery by enzyme digestion and divided into 3 groups: group 1 and 2 were cultured in the presence of DMEM/F12 containing either FBS or UCP; and group 3 was cultured in serum-free medium (StemPro® MSC SFM CTS™). Morphology, proliferation and surface marker expression were examined by flow cytometry, and the differentiation toward adipogenic and osteogenic lineages was used for investigating the effect of media on UCMSC after 3-5 passages. Next, the cells cultured in the three different media were cryopreserved and thawed, then prepared as feeder layers with the name of UCMSC(FBS), UCMSC(UCP), and UCMSC(SFM), respectively. The CD34⁺ cells were separated from UCB by magnetic activated cell sorting (MACS) and divided into 4 groups cultured in StemPro(-34) SFM medium added with hematopoietic cytokine combination (StemSpan® CC100). The control group included only CD34⁺ cells as group A (blank control) and experimental groups included UCMSC(FBS) + CD34⁺ cells as group B, UCMSC(UCP) + CD34⁺ cells as group C, UCMSC(SFM) + CD34⁺ cells as group D, and cells in all groups were cultured ex vivo for 7 days. The nucleated cell (NC) number was counted by cell counter, CD34⁺ cells were measured by flow cytometry, and clonogenic assay was conducted at day 0 and 7 of culture. The expansion efficiency was assessed.</p><p><b>RESULTS</b>The morphology (spindle-shaped and plastic-adherent), the immunophenotype (high positive percentage of CD73, CD90, CD105 and CD166) and the differentiation potential (osteogenic and adipogenic) were almost indistinguishable among the cells cultured in any of these three media except for the expression of CD105 in group 3 (serum-free medium) was lower than that in other 2 groups (P < 0.05). UCMSC grown in UCP medium demonstrated significantly higher proliferation rates than that in media containing FBS or commercial serum-free supplement (P < 0.05). After co-culture for 7 days, the CD34⁺ cell percentage decreased in all the groups, while NC were amplified effectively and the CD34⁺ cell number increased with the same order as group C or D group B or A (control group) (P < 0.05). As compared with the colony-forming unit (CFU) number at day 0, there was no significant difference in the expansion multiple between group C and D, while the expansion of CFU in group C were higher than that in group B and A.</p><p><b>CONCLUSION</b>The UCP can be used as a better animal-free serum supplement for growth, maintenance and differentiation of UCMSC, thus would be a safe choice for clinical-scale production of human MSC.</p>


Assuntos
Humanos , Antígenos CD34 , Biomarcadores , Técnicas de Cultura de Células , Diferenciação Celular , Separação Celular , Células Cultivadas , Técnicas de Cocultura , Sangue Fetal , Citometria de Fluxo , Imunofenotipagem , Células-Tronco Mesenquimais , Cordão Umbilical
5.
Artigo em Chinês | WPRIM | ID: wpr-265717

RESUMO

<p><b>OBJECTIVE</b>To investigate the effects of hypertonic sodium chloride hydroxyethyl starch 40 (HSH) on brain edema and morphological changes during whole body hyperthermia (WBH) in rats.</p><p><b>METHODS</b>Sixty adult male SD rats were randomized into control group, WBH group without fluid infusion (group HT), WBH group with Ringer's infusion (group RL), WBH group with HAES + Ringer's infusion (group HRL) and WBH group with HSH infusion (group HSH). WBH was induced by exposure to 36 degrees celsius; for 3 h to achieve a rectal temperature of 41-42 degrees celsius;, and the corresponding fluids were administered intravenously within 30 min at the beginning of WBH. The control rats were housed at a controlled room temperature (22∓1) degrees celsius; for 4 h. After cooling at room temperature for 1 h, the rats were sacrificed and brain water content and morphological changes were evaluated.</p><p><b>RESULTS</b>Compared with the control group, all the WBH groups had significantly increased brain water content (P<0.05 or 0.01), but group HSH showed a significantly lower brain water content than group HT (P<0.05). The rats in groups HT, RL and HRL showed serious to moderate structural changes of the brain tissue and nerve cells, but HSH group had only mild pathologies.</p><p><b>CONCLUSION</b>HSH can reduce brain edema and ameliorate the damages to brain cells in rats exposed to WBH.</p>


Assuntos
Animais , Masculino , Ratos , Encéfalo , Patologia , Edema Encefálico , Patologia , Derivados de Hidroxietil Amido , Usos Terapêuticos , Hipertermia Induzida , Ratos Sprague-Dawley , Solução Salina Hipertônica , Usos Terapêuticos
6.
Chin. med. j ; Chin. med. j;(24): 856-861, 2011.
Artigo em Inglês | WPRIM | ID: wpr-239935

RESUMO

<p><b>BACKGROUND</b>Omeprazole, usually used in the antiplatelet therapy during percutaneous coronary intervention (PCI) in acute coronary syndrome (ACS), has been reported to increase ischemic events in retrospective studies. However, other clinical trials gave paradoxical results. The aim of this study was to assess the effects of omeprazole on clopidogrel efficacy and clinical events.</p><p><b>METHODS</b>All patients (n = 172) received aspirin (loading dose 300 mg and maintenance dose 100 mg/d) and clopidogrel (loading dose 600 mg and maintenance dose 75 mg/d) during the therapy. They were randomized to receive omeprazole (20 mg/d) or placebo for 30 days. Residual platelet activities in the adenosine 5'-diphosphate (ADP) pathway were detected on the fifth day after PCI with thrombelastography (TEG)-mapping. The clinical events were recorded after one month.</p><p><b>RESULTS</b>According to the five levels of platelet activities, the frequency distributions of the inhibition rates were significantly different (P = 0.0062). However, no significant change was seen in the distribution among the highest or the lowest inhibiting levels (> 95% and < 30% inhibition rate). And there were no significant differences (P > 0.05) in events incidence, while gastro-intestinal bleeding decreased in co-administration of omeprazole.</p><p><b>CONCLUSIONS</b>Omeprazole significantly blunts clopidogrel efficacy while not exacerbates ischemic events in ACS undergoing PCI. Omeprazole even can decrease gastro-intestinal bleeding in those patients.</p>


Assuntos
Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Síndrome Coronariana Aguda , Sangue , Tratamento Farmacológico , Patologia , Terapêutica , Angioplastia Coronária com Balão , Métodos , Aspirina , Usos Terapêuticos , Omeprazol , Usos Terapêuticos , Inibidores da Agregação Plaquetária , Usos Terapêuticos , Ticlopidina , Usos Terapêuticos
7.
Artigo em Chinês | WPRIM | ID: wpr-313901

RESUMO

This study was purposed to investigate the role of post-thaw infused donor cells for predicting engraftment and hematopoietic reconstitution after unrelated cord blood transplantation (UCBT). The retrospective analysis was performed on clinical data of 97 children with malignant or non-malignant diseases received single unit UCBT from August 1999 to April 2010. The impact of pre-freezing and post-thaw cell dose of total nucleated cells (TNC), CD34(+) cells and colony-forming units-granulocyte/macrophage (CFU-GM) on engraftment and hematological recovery after UCBT was analyzed. Unrelated donors were from Guangzhou cord blood bank (GZCBB) entirely. The results indicated that the pre-freezing TNC (/kg) (mean ± SD: 7.65 × 10⁷ ± 4.26 × 10⁷; median: 6.34 × 10⁷), CD34(+)cells (/kg) (mean ± SD: 4.64 × 10(5) ± 4.47 × 10⁵; median: 3.03 × 10⁵) and CFU-GM (/kg) (mean ± SD: 0.79 × 10⁵ ± 1.09 × 10⁵; median: 0.57 × 10⁵) showed a good correlation with their post-thaw counterparts including TNC(/kg) (mean ± SD: 6.98 × 10⁷ ± 4.12 × 10⁷; median: 6.00 × 10⁷), CD34(+)cells (/kg)(Mean ± SD: 6.86 × 10⁵ ± 8.56 × 10⁵; Median: 4.17 × 10⁵), and CFU-GM (/kg) (mean ± SD: 0.52 × 10⁵ ± 0.52 × 10⁵; median: 0.39 × 10⁵) (r = 0.952, p < 0.001; r = 0.794, p < 0.001; r = 0.478, p < 0.001). Either the pre-freezing or post-thaw number of infused CFU-GM was significant higher in patients who achieved engraftment (n = 70) than those who suffered graft failure (n = 22) (p = 0.023 and 0.011, respectively), but no significant difference of TNC and CD34(+) cells dose (pre-freezing or post-thaw) were found between these two groups. Pre-freezing CFU-GM, TNC, CD34(+) cell dose negatively correlated with the time of neutrophil engraftment (r = -0.285, p = 0.018; r = -0.396, p = 0.002; r = -0.373, p = 0.002), as well as the post-thaw number of TNC and CD34(+) cells (r = -0.260, p = 0.031; r = -0.483, p < 0.001), whereas only pre-freezing CD34(+) cells showed a significant correlation with platelet engraftment time (r = -0.352, p = 0.013). It is concluded that the CFU-GM amount is useful for predicting engraftment of UCBT, while pre-freezing hematopoietic cell doses show superior correlation with the speed of engraftment and hematopoietic reconstitution than their post-thaw counterparts in pediatric recipients, suggesting that it is essential to perform hematopoietic potency assay on each cord blood unit prior to listing or release for administration.


Assuntos
Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Antígenos CD34 , Sangue , Bancos de Sangue , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Métodos , Sangue Fetal , Biologia Celular , Sobrevivência de Enxerto , Células Progenitoras de Granulócitos e Macrófagos , Estudos Retrospectivos , Doadores de Tecidos
8.
Journal of Experimental Hematology ; (6): 1535-1541, 2010.
Artigo em Chinês | WPRIM | ID: wpr-332323

RESUMO

Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells for transplantation with success being associated with the total nucleated cell (TNC) count, CD34(+) cells and colony-forming unit-granulocyte-macrophage (CFU-GM) content infused. This study was purposed to clarify the impact of maternal and neonatal factors on hematopoietic potential of UCB product. UCB samples were screened, processed, tested and cryopreserved according to the Standard Operation Procedure (SOP) of Guangzhou cord blood bank (GZCBB). Relationship of hematopoietic cell parameters with maternal and neonatal characteristics for 4615 UCB units was analyzed retrospectively. The results showed that both collected volume (Mean ± SD: 95.23 ± 22.42 ml; Median: 91.85 ml) and initial TNC [Mean ± SD: (1.34 ± 0.49) × 10(9); Median: 1.25 × 10(9)] correlated well with postprocessed TNC [Mean ± SD: (1.21 ± 0.42) × 10(9); Median: 1.14 × 10(9); p < 0.001], CD34(+)count [Mean ± SD: (5.14 ± 4.55) × 10(6); Median: 4.08 × 10(6); p < 0.001] and CFU-GM content [Mean ± SD: (9.72 ± 8.66) × 10(5); Median: 7.53 × 10(5); p < 0.001]. As for donor factors, only infant birth weight correlated strongly with volume collected and all hematopoietic cell parameters (p < 0.001). UCB samples from bigger babies had higher collected volume, TNC, CD34(+) count and CFU-GM content (p < 0.001). Mother's age had no correlation with all the above parameters. Gestational age correlated positively with initial/postprocessed TNC (p < 0.001) and negatively with CD34(+) count (p = 0.04), but no relation with collected volume and CFU-GM content. Cesarean section produced superior volume (Mean ± SD: 97.05 ± 22.23 ml vs 92.53 ± 22.43 ml; Median: 94.08 ml vs 88.82 ml; p < 0.001), but inferior cell count than vaginal delivery (p < 0.001). Male infants had more initial volume and CD34(+) count (Mean ± SD: 96.41 ± 22.31 ml vs 93.95 ± 22.47 ml; Median: 93.27 ml vs 90.14 ml; p < 0.001); [Mean ± SD: (5.28 ± 5.04) × 10(6) vs (5.00 ± 3.94) × 10(6); Median: 4.18 × 10(6) vs 3.94 × 10(6); p < = 0.042], but lower initial and postprocessed TNC than female ones [Mean ± SD: (1.31 ± 0.50) × 10(9) vs (1.37 ± 0.47) × 10(9); Median: 1.22 × 10(9) vs 1.28 × 10(9); p < 0.001]; [Mean ± SD: (1.18 ± 0.42) × 10(9) vs (1.24 ± 0.41) × 10(9); Median: 1.10 × 10(9) vs 1.17 × 10(9); p < 0.001], while no significant difference of CFU-GM were found between male and female infants. It is concluded that these data may be helpful to optimize the UCB donor selection and improve cost efficiency of UCB bank resource. The heavier infants after vaginal delivery should be selected and large-volume units with higher TNC should be chosen at first.


Assuntos
Adulto , Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Adulto Jovem , Peso ao Nascer , Bancos de Sangue , Métodos , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Métodos , Parto Obstétrico , Seleção do Doador , Sangue Fetal , Biologia Celular , Alergia e Imunologia , Idade Gestacional , Células-Tronco Hematopoéticas , Idade Materna
9.
Biomed. environ. sci ; Biomed. environ. sci;(12): 78-83, 2007.
Artigo em Inglês | WPRIM | ID: wpr-249885

RESUMO

<p><b>OBJECTIVE</b>To investigate the function of a novel biosensor used for the rapid determination of biochemical oxygen demand (BOD) which is developed by our research group based on suspended immobilized microbial cell system in a completely mixed determining chamber as a substitute of the traditional membrane system.</p><p><b>METHODS</b>Activated sludge was immobilized by PVA gel and used as a bio-sensing element. The novel biosensor was used to measure the short time BOD value and the conventional cultivation method was used for BOD5 measurement.</p><p><b>RESULTS</b>A linear relationship was observed for the difference between the current and the concentration of glucose-glutamic acid (GGA) solution below 200 mg/L with a correlation coefficient of 0.995. The optimal response of the sensor was obtained at pH 7.0 and 30 degrees C. The sensor response was within 15 min and was reproducible within +/- 5% of the mean in a series of eight samples containing 75 mg/L BOD using standard GGA solution. The novel sensor response was found to be fairly constant over a period of 30 days, with +/- 5% fluctuations.</p><p><b>CONCLUSION</b>A relatively good agreement is found between BOD estimated by the novel BOD biosensor and that determined by the conventional 5-day BOD method. This novel BOD biosensor has good sensitivity, stability and reproducibility.</p>


Assuntos
Técnicas Biossensoriais , Ácidos Bóricos , Química , Glucose , Metabolismo , Ácido Glutâmico , Metabolismo , Oxigênio , Metabolismo , Álcool de Polivinil , Química , Reprodutibilidade dos Testes , Esgotos , Microbiologia , Temperatura
10.
Artigo em Chinês | WPRIM | ID: wpr-230307

RESUMO

This study was purposed to investigate the immunoregulatory effect of endothelial cells derived from mesenchymal stem cells (MSC). The human MSC was induced to differentiate into endothelial cells for one week. The phenotypes were evaluated by flow cytometry, the cell morphologic feature was observed by invert phase-contrast microscope and analysis of capillary formation was performed by using the in vitro angiogenesis kit. The immunoregulatory effect was detected by lymphocyte transformation test. The result indicated that during the differentiation cells grew fast and there was no significant change in the phenotypes, i.e. CD73, CD105, HLA-ABC were positive and CD34, CD80, CD86, HLA-DR, CD31 were negative. Immunofluorescence analysis showed typical expression of the von Willebrand factor. Differentiated MSCs formed capillary-like structure. Endothelial cells derived from MSC also revealed immunosuppressive effect on T cell proliferation in a dose-dependent manner. It is concluded that endothelial cells derived from MSC also harbor immunoregulatory effect on T lymphocytes.


Assuntos
Criança , Humanos , 5'-Nucleotidase , Metabolismo , Diferenciação Celular , Fisiologia , Células Cultivadas , Células Endoteliais , Biologia Celular , Alergia e Imunologia , Células-Tronco Mesenquimais , Biologia Celular , Metabolismo , Linfócitos T , Alergia e Imunologia , Fator de von Willebrand , Metabolismo
11.
Artigo em Chinês | WPRIM | ID: wpr-282758

RESUMO

From June 1998 to July 2004, Guangzhou umbilical cord blood bank provided unrelated umbilical cord blood for 54 patients to more than 21 transplantation centers. HLA sequencing-based typing (SBT) was used to re-analyze the results of HLA antigens and alleles so as to investigate the relationship between HLA alleles and GVHD. The information about 48 out of 54 patients was obtained after 6 months of follow up. SBT was used to identify HLA-A, B, DRB1 alleles in 48 patients received the unrelated umbilical cord blood units, and the obtained results were compared with the results of HLA-SSP Low Resolution Typing. The results showed that the difference of GVHD incidence between less than 2 mismatched HLA sites and less than 3 sites was statistically significant (P < 0.05). In the results from single factor analysis and high-resolution typing of HLA-A, B and DRB1 alleles, the mismatch between HLA-B and HLA-DRB1 alleles was found to be a significant factor for the occurence of GVHD. It is concluded that SBT plays an important role in umbilical cord blood transplantation, and the incidence of GVHD is higher in the transplantation with HLA-DRB1 alleles mismatching.


Assuntos
Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Alelos , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Sangue Fetal , Biologia Celular , Alergia e Imunologia , Doença Enxerto-Hospedeiro , Antígenos HLA-A , Genética , Alergia e Imunologia , Antígenos HLA-B , Genética , Alergia e Imunologia , Antígenos HLA-DR , Genética , Alergia e Imunologia , Cadeias HLA-DRB1 , Teste de Histocompatibilidade , Métodos , Leucemia , Terapêutica , Análise de Sequência
12.
Artigo em Chinês | WPRIM | ID: wpr-233546

RESUMO

The study was aimed to establish a protocol of isolating and culturing adult mesenchymal stem cells (MSC) from human bone marrow aspirate and identify them by surface antigen analysis and committed differentiation in order to provide an experimental foundation for achieving a therapeutic benefit in applying MSC in hematopoietic stem cell transplantation. MSCs were obtained from fresh human bone marrow aspirate by gradient centrifugation with Percoll (1.073 g/ml) and anchoring culture in L-DMEM with 10% fetal bovine serum by a full medium exchange every 3 days. The MSC surface antigens, including CD34, CD45, CD73, CD105, CD166, were analyzed on FACScan flow cytometer. Under culture in conditioned medium for osteogenesis (the hormone cocktail containing 0.1 micromol/L dexamethasone, 10 mmol/L glycerol-2-phosphate and 50 micromol/L ascorbic acid) and adipogenesis (the cocktail containing 1 micromol/L dexamethasone, 5 mg/L insulin, 0.5 mmol/L 1-methyl-3-isobutylxanthine and 60 micromol/L indomethacin), MSCs committedly differentiated into osteoblasts and adipocytes. The differentiated mesenchymal stem cells were identified by morphological observation and immunohistochemical staining. The results showed that by gradient centrifugation and adhesion culture, MSCs could be isolated and culture-expanded from human bone marrow aspirate. These cells were uniformly negative for CD34, CD45 and positive for CD73, CD105 and CD166. The osteogenic differentiated cells were positive for alkaline phosphatase (ALP) and the adipogenic differentiated cells displayed accumulation of lipid vacuoles, as detected by oil red O. It is concluded that MSC can be isolated and expand-cultured from adult human bone marrow aspirate and committedly differentiate into osteoblasts and adipocytes. MSC primary identification can be accomplished by flow cytometry and induced differentiation. The set of methods in current experiment shows somewhat practical value for basic research and clinical application.


Assuntos
Humanos , 5'-Nucleotidase , Metabolismo , Antígenos CD , Metabolismo , Células da Medula Óssea , Biologia Celular , Moléculas de Adesão Celular Neuronais , Metabolismo , Técnicas de Cultura de Células , Diferenciação Celular , Fisiologia , Separação Celular , Métodos , Endoglina , Proteínas Fetais , Metabolismo , Células-Tronco Mesenquimais , Biologia Celular , Receptores de Superfície Celular , Metabolismo
13.
Chinese Journal of Pediatrics ; (12): 220-223, 2006.
Artigo em Chinês | WPRIM | ID: wpr-309241

RESUMO

<p><b>OBJECTIVE</b>From December 1998 to April 2004, 3960 umbilical cord blood units were stored in Guangzhou cord blood bank, which provided 100 umbilical cord blood units to 25 transplant center for 83 patients with malignant or non-malignant diseases. To study the related factors affecting unrelated umbilical cord blood stem cell transplantation, the authors analyzed retrospectively the results of transplantation of unrelated umbilical cord blood stem cells for 65 patients.</p><p><b>METHODS</b>ALL (acute lymphocytic leukemia) cord blood units were obtained from full term normal vaginal and cesarean deliveries in Guangzhou Women and Infants Hospital. The fractionation, cryopreservation and thawing of the cord blood were performed according to the regulations of New York umbilical cord blood bank and pertinent literature. The selection of cord blood was based on HLA typing and the number of nucleated cells. The sex and HLA antigens of donors were defined as the evidence of engraftment. Time to engraftment was recorded when the absolute number of neutrophil ANC (absolute neutrophil count) was higher than 5.0 x 10(8) for three days. Event-free survival and graft versus host disease (GVHD) were provided by transplant centers.</p><p><b>RESULTS</b>Out of 65 patients who received unrelated cord blood stem cell transplant, 49 patients were diagnosed as having malignant diseases [including 23 with ALL, 16 with AML (acute myeloid leukemia), 7 with CML (chronic myelogenous leukemia), 3 with lymphoma and one with MDS (myelodysplastic syndrome)], 16 patients had non-malignant disease. The 65 transplanted patients (42 male, 23 female) had a median age of 10 years (range 1 - 33 years) and a median body weight of 27 kg (range 10 - 67 kg). The patients received cord blood stem cells from unrelated 0-locus (n = 9) or 1-locus (n = 43) or 2-locus (n = 13) HLA mismatched donor. The median dose of infused cells was: total neutrophil count (TNC) 5.7 x 10(7), CD(34)(+) 5.1 x 10(5), CFU-GM 3.8 x 10(4). Fifty of 65 (77%) patients had engraftment. GVHD occurred in 41 patients (63%), including acute grade I - II GVHD in 31 patients (76%), acute grade III - IV GVHD in 8 patients (20%) and chronic GVHD in 2 patients (5%). Fifty patients had engraftment (ANC > 5.0 x 10(8)) after a median time of 17 (range 7 to 44) days after transplant, while an autologous hematopoietic reconstitution was observed in 6 patients; 24 patients died of severe pneumonia (n = 8), acute GVHD (n = 4), or sepsis (n = 12) and the disease-free survival probability was 61%.</p><p><b>CONCLUSIONS</b>Unrelated allogeneic umbilical cord blood transplantation may be a good substitution for unrelated allogeneic bone marrow transplantation with a good prospect.</p>


Assuntos
Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Adulto Jovem , China , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Métodos , Intervalo Livre de Doença , Doença Enxerto-Hospedeiro , Mortalidade , Leucemia , Mortalidade , Terapêutica , Estudos Retrospectivos , Taxa de Sobrevida , Transplante Homólogo , Resultado do Tratamento
14.
Zhonghua Wai Ke Za Zhi ; (12): 259-262, 2005.
Artigo em Chinês | WPRIM | ID: wpr-264528

RESUMO

<p><b>OBJECTIVE</b>To study sagittal mobility about the FTJ (first tarsometatarsal joint) and its relationship with the pathophysiology and treatment of hallux valgus patients.</p><p><b>METHODS</b>According to Lee's method, FTJ sagittal mobility of 300 normal feet and 200 hallux valgus was measured, and its correlative factors were statistically analysed.</p><p><b>RESULTS</b>FTJ sagittal mobility of 300 normal feet was 8.4 degrees +/- 2.3 degrees , and that of 200 hallux valgus was 11.7 degrees +/- 3.2 degrees , the difference was significant. The normal range of FTJ sagittal mobility was less than 13 degrees . The sagittal overmotion of FTJ had relation to the anatomical configuration of FTJ (P < 0.05), intercuneiform splitting (P < 0.01), transferred pain under the second metatarsal head (P < 0.01), and FTJ osteoarthritis (P < 0.01) had no relation to HVA (hallux valgus angle), IMA (intermetatarsal angle), second metatarsus medial diaphyseal cortex hypertrophy (P > 0.05).</p><p><b>CONCLUSION</b>Lee's method is convenient and accurate. Both HVA and IMA can not represent the sagittal mobility measurement of FTJ, which should be routinely evaluated, especially for hallux valgus patients with type I FTJ. Lapidus procedure should be considered for patients with larger FTJ in combination with transferred pain under the second metatarsal head, intercuneiform splitting, FTJ osteoarthritis.</p>


Assuntos
Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Hallux Valgus , Diagnóstico , Cirurgia Geral , Articulação Metatarsofalângica , Fisiologia , Cirurgia Geral , Osteotomia , Métodos , Amplitude de Movimento Articular
15.
Artigo em Chinês | WPRIM | ID: wpr-356585

RESUMO

In order to research the related factors of umbilical cord blood transplantation, 54 cases of unrelated umbilical cord blood transplantation were analyzed retrospectively, which were performed from June 1998 to July 2003. All cord blood units were obtained from full term normal vaginal deliveries in Guangzhou Maternal-Neonatal Hospital. The fractionation, cryopreservation and thawing of cord blood have been done according to the regulation of New York umbilical cord blood bank and pertinent literature. The selection of cord blood is based on HLA typing and the number of nucleated cells. The results showed that from June 1998 to July 2003, 3 475 units of cord blood were collected in Guangzhou Umbilical Cord Blood Bank and 99 units were provided for therapy of 85 patients in 21 transplantation centers, including 11 sibling and 74 unrelated cord blood transplantations. 54 cases of unrelated cord blood transplantation were reported, including 43 malignant diseases and 11 non-malignant diseases. The median age of recipients was 9.5 (range 1.2 - 33) years, the median weight was 27 (range 10 - 60) kg, the median number of TNC was 6.82 x 10(7)/kg, 43 cord blood were implanted (ANC > 500/microl) at day 60 after transplantation (79.6%, median 17). The time of nuclear cell reconstitution after cord blood transplantation was statistically related with nucleated cells and the type of disease, not related with HLA matching. Acute GVHD was present in 8 patients (21.6%) and chronic GVHD occurred in 2 patients (5.4%), 6 patients suffered from graft failure (11.1%). The total survival rate was 42.6%. It is suggested that unrelated umbilical cord blood transplantation seems to be a good substitute for bone marrow transplantation and has good prospects especially in children.


Assuntos
Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Doença Aguda , China , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Métodos , Doença Enxerto-Hospedeiro , Mortalidade , Leucemia , Cirurgia Geral , Estudos Retrospectivos , Taxa de Sobrevida
16.
Artigo em Chinês | WPRIM | ID: wpr-278870

RESUMO

The objective of this research was to explore whether the number of CD34(+)CD38(+) cells infused affects hematopoietic reconstitution after cord blood transplantation. The number of CD34(+)CD38(+) cells in cord blood was analysed with flow cytometry after freezethawing. The body weight and time for neutrophil and platelet recovery were measured in 20 children with acute leukemia. The results showed that the median number of CD34(+)CD38(+) cells infused was 29.47 (9.85 - 325.71) x 10(4)/kg. A median time for neutrophil recovery (> 5 x 10(8)/L) in 20 patients was 18.5 (11 - 32) days, and time for platlet recovery (> 2 x 10(10)/L) in 19 of 20 patients was 45 (12 - 118) days. The number of CD34(+)CD38(+) cells infused correlated with time to neutrophil and platelet recovery (r = -0.577, P < 0.01 and r = 0.503, P < 0.05, respectively). In conclusion, the number of CD34(+)CD38(+) cells infused is correlated with the time for hematologic recovery.


Assuntos
Adolescente , Criança , Pré-Escolar , Humanos , ADP-Ribosil Ciclase , ADP-Ribosil Ciclase 1 , Antígenos CD , Antígenos CD34 , Sangue Fetal , Biologia Celular , Transplante , Hematopoese , Transplante de Células-Tronco Hematopoéticas , Métodos , Leucemia Mieloide Aguda , Sangue , Terapêutica , Glicoproteínas de Membrana , Leucemia-Linfoma Linfoblástico de Células Precursoras , Sangue , Terapêutica
17.
Chinese Journal of Burns ; (6): 355-357, 2003.
Artigo em Chinês | WPRIM | ID: wpr-352254

RESUMO

<p><b>OBJECTIVE</b>To observe the injury on micro-skin induced by a self designed micro-skin machine.</p><p><b>METHODS</b>Micro-skin was produced either with the machine or by hand. Cells at the edge of micro-skin were observed by transmission electron microscope. succinic dehydrogenase activity in supernatant of cultivated cells was analyzed, and the cell proliferation of micro-skin was assessed by (3)H-TdR. Twenty patients were enrolled in the study for the observation of the wound healing time between the two groups of micro-skin after being grafted.</p><p><b>RESULTS</b>Transmission electron microscope examination revealed that the cellular injury at the edge of the micro-skin in machine-made group was mild compared with that in man-made group. (3)H-TdR rate was elevated but the activity of succinic dehydrogenase in the supernatant of cultured cells decreased in supernatant of cultured cells of machine produced micro-skin. Wound healing time was shortened in machine made group. (P < 0.05).</p><p><b>CONCLUSION</b>The cellular injury at the edge of micro-skin in the machine made group was mild when compared with that in the man-made group with cell proliferation accelerated and wound healing time shortened.</p>


Assuntos
Humanos , Queimaduras , Cirurgia Geral , Divisão Celular , Epitélio , Patologia , Microscopia Eletrônica , Pele , Transplante de Pele , Métodos , Cicatrização
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