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Objective To investigate the role of lncRNA SNHG1 in homocysteine-induced pyroptosis of podocyte.Methods Cbs+/-mice were randomly divided into two groups:a normal diet group(ND)and a high me-thionine diet group(HMD).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocytes were cultured in vitro,and then assigned into a control group(Control,0 μmol/L Hcy)and a homocysteine intervention group(Hcy,80 μmol/L Hcy).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocyion group(OE-NC + Hcy)and the lncSNHG1 overexpression + homocysteine intervention group(OE-SNHG1 + Hcy)were also established.After 48 hours of intervention,Real-time fluorescence quantita-tive PCR was used to detect the expression of lncSNHG1 in podocytes after Hcy intervention.Western blot was used to detect the expressions of Caspase-1,Cleaved Caspase-3 and NLRP3.Immunofluorescence was used to de-tect the expression levels of GSDMD and GSDMD-N.ELISA was used to detect the contents of IL-1β and IL-18.Results(1)In the animal experiments,the expression levels of pyroptosis-related proteins Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the HMD group compared with the ND group.(2)In the cellular experiments,the expression levels of Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the Hcy group compared with the Control group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased as well.(3)In the cellular experiments,the expres-sion of lncSNHG1 was increased in the Hcy group compared with the control group.After transduction with lnc-SNHG1 lentivirus,the expression of lncSNHG1 was increased in the OE-SNHG1 group,compared with the control group and the OE-NC group.(4)In the cellular experiments,the expressions of pyroptosis-related proteins Cas-pase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were increased compared with the OE-NC+Hcy group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased in the OE-SNHG1+Hcy group.Conclusion These results indicate that lncSNHG1 may play a role in promoting Hcy induced podocytepyroptosis.
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This study discussed the haplotype characteristics of mitochondrial control region (D-loop region) of broiler breeds with different growth rates as well as the relationships between different haplotypes and growth performance. The D-loop region's sequence size of the 974 individuals from 20 broiler breeds ranged from 1,231 to 1,232bp, as a C base deletion at 859 bp. A total of 52 mutation sites and 72 haplotypes were detected, which were divided into four haplogroups, A, B, C and E. Of these, haplogroup E was the dominant haplogroup among 817 broilers and all medium/fast-growing broiler breeds. While its proportion was not exceed 40.00% among others types breeds. Correlation analysis showed that there was a significant negative correlation between birth weight and haplogroup A and a significant positive correlation between birth weight and haplogroup E. Age in days and feed conversion ratios were positively correlated with haplogroup B but negatively correlated with haplogroup E when the average body weight was approximately 1.8 kg. The cluster analysis showed that haplogroups A and B with Gallus gallus spadiceus and E with Gallus gallus murgha, respectively clustered into one branch. This study provided a theoretical basis for broiler breeding and genetic resource protection, development and utilization.
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Galinhas , Variação Genética , Humanos , Animais , Haplótipos/genética , Galinhas/genética , DNA Mitocondrial/genética , Peso ao Nascer/genética , Cruzamento , FilogeniaRESUMO
Objective To investigate the effects of Loropetalum chinense extracts on the proliferation of lung adenocarcinoma A549 cells cultured in vitro. Methods CCK-8 assay was performed to evaluate the effect of Loropetalum chinense extracts on the proliferation of A549 cells, and the clonal growth ability of A549 cells was determined by clone formation assay. Flow cytometry Annexin V-APC/PI double staining was used to measure the apoptosis of A549 cells. Western blot was used to measure the expressions of apoptosis-related proteins Bax, Fas, Bcl-2 Caspase3 and cleaved-Caspase 3. Results Loropetalum chinense extracts significantly inhibited the proliferation and colony formation of A549 cells in a time- and dose-dependent manner. The viability of A549 cells decreased by 50% after 48h treatment of 0.5mg/ml extracts, and 100% inhibition of colony formation rate achieved when the cells were treated with 40μg/ml extracts for 14 days. When A549 cells were treated with Loropetalum chinense extracts for 24h, apoptotic rates increased in a dose-dependent manner. Compared with the control group, the protein levels of Fas, Bax, Caspase3 and cleaved-Caspase3 were up-regulated, while Bcl-2 was down-regulated. Conclusion Loropetalum chinense extracts inhibit the growth of human lung adenocarcinoma cells in vitro, and the mechanisms may be related to the activation of mitochondrial and death receptor apoptosis pathway.
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Objective:To summarize the classification of etiology, age of onset, prognosis of children with convulsion, so as to provide experience guidance for clinicians engaged in pediatric emergency department.Methods:The clinical data of children with convulsions received in the emergency department of Children′s Hospital Affiliated to Kunming Medical University from January 2015 to December 2018 were analyzed retrospectively.Results:During the four-year period, 2 957 children with convulsion were received in the emergency department, accounting for 22.20% of the total number of critically ill children in the observation room of the emergency department, and the ratio of male to female was 1.7∶1.The etiological diagnosis of convulsion in emergency are as follows: febrile convulsion(733 cases, 24.79%), central nervous system infection(477 cases, 16.13%), unexplained convulsion(476 cases, 16.09%), epilepsy(371 cases, 12.55%), benign infantile convulsions with mild gastroenteritis(240 cases, 8.12%). The age of onset: 8.25% were in neonatal period, 33.99% were in infant, 34.87% were in toddler′s age, 12.17% were in preschool age, 7.88% were in school age and 2.84% were in adolescence.Destination statistics: 72.00% were admitted to hospital for further treatment, 13.29% were transferred to neurology clinic, 7.85% to pediatric clinic, 1.66% to rehabilitation clinic, and 0.17% died.Inpatient department: 43.64% were admitted to department of neurology, 17.52% to pediatric intensive care unit, 13.71% to department of neonatology, 12.64% to department of gastroenterology and 2.72% to department of rehabilitation.Conclusion:Febrile convulsion is the main cause of convulsion in children who were received emergency treatment in our hospital.Most of the convulsion cases are from birth to preschool age, and the prognosis is good after active treatment.
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Objective To investigate the changes of serum creatine kinase (CK),creatine kinase isoenzyme (CK-MB),cardiac troponin I(cTnI)and electrocardiogram (ECG) before and after the treatment of pneumo-nia in children.Methods From December 2014 to December 2016,95 children with pneumonia were selected as the study group,and 48 healthy subjects who underwent the healthy assessment from December 2014 to January 2016 were selected as the control group.All children with pneumonia were treated after admission.2 mL of venous blood were collected from each research subject after the admission and patients in study group after treatment,serum was seperated,and levels of CK,CK-MB,cTnI were measured and the ECG record was conducted.Results The serum levels of CK,CK-MB and cTnI in the study group were higher than those in the control group (P<0.05);the incidences of ST segment elevation or depression,atrial premature beat,ven-tricular premature beat,sinus tachycardia and sinus bradycardia in the study group were higher than those in the control group,and the differences were statistically significant (P<0.05);the serum levels of CK,CK-MB and cTnI in the study group were lower than those before treatment,and the difference was statistically signif-icant (P<0.05);the incidences of atrial premature beat,ventricular premature beat,sinus tachycardia and si-nus bradycardia in the study group after treatment were lower than those before treatment,and the difference was statistically significant (P<0.05);the incidence of ST segment elevation or depression after treatment in the study group was lower than that before treatment,and there was no significant difference (P>0.05).Con-clusion The serum levels of CK,CK-MB,cTnI and ECG were obviously abnormal in children with pneumoni-a.After treatment,serum CK,CK-MB and cTnI levels can be reduced and ECG abnormalities can be ameliora-ted.
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Glioma is an aggressive malignancy with limited effective treatment and poor prognosis. Cytoplasmic polyadenylation element binding protein 4 is a regulator of gene transcription and has been reported to be associated with biological malignancy in cancers. However, the mechanisms that cytoplasmic polyadenylation element binding protein 4 contributes to tumor migration and invasion remain unknown. Here, cytoplasmic polyadenylation element binding protein 4 expression was assessed using immunohistochemistry, and the results were compared with clinicopathological parameters, including survival. Using glioma cell lines (SKMG-4 and T98G), we measured cytoplasmic polyadenylation element binding protein 4 messenger RNA and protein expression and studied the effects of cytoplasmic polyadenylation element binding protein 4 expression on cell migration and invasion. Cytoplasmic polyadenylation element binding protein 4 expression was significantly higher in tumor tissues than that in normal brain tissues. Clinicopathological analysis showed that cytoplasmic polyadenylation element binding protein 4 expression was significantly correlated with advanced World Health Organization grade ( p < 0.001) and lower Karnofsky Performance Status (KPS) score ( p = 0.001). Cytoplasmic polyadenylation element binding protein 4 positive as opposed to the cytoplasmic polyadenylation element binding protein 4 negative patients had lower overall survival ( p < 0.001). Multivariate analysis suggested that cytoplasmic polyadenylation element binding protein 4 expression might be an independent prognostic indicator (hazard ratio = 2.091, 95% confidence interval: 1.093-3.999, p = 0.026) for glioma patients. Moreover, upregulated cytoplasmic polyadenylation element binding protein 4 expression could promote T98G cell migration and invasion, and downregulated cytoplasmic polyadenylation element binding protein 4 expression could inhibit SKMG-4 cell migration and invasion. Furthermore, downregulated cytoplasmic polyadenylation element binding protein 4 could reduce the protein expression of matrix metalloproteinase-2 and matrix metalloproteinase-9. In conclusion, our studies indicated that positive cytoplasmic polyadenylation element binding protein 4 expression predicted a worse prognosis in glioma patients, and cytoplasmic polyadenylation element binding protein 4 could represent a useful biomarker or therapeutic target for glioma.
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Neoplasias Encefálicas/patologia , Movimento Celular , Glioma/patologia , Proteínas de Ligação a RNA/fisiologia , Adulto , Idoso , Neoplasias Encefálicas/mortalidade , Linhagem Celular Tumoral , Feminino , Glioma/mortalidade , Humanos , Masculino , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 9 da Matriz/análise , Pessoa de Meia-Idade , Invasividade Neoplásica , Prognóstico , Proteínas de Ligação a RNA/análiseRESUMO
BACKGROUND:Chitosan biological materials can induce bone marrow mesenchymal stem cells to differentiate toward neurons. As a derivative of chitosan, carboxymethyl chitosan has a series of excelent properties. However, whether carboxymethyl chitosan can induce the neuronal differentiation of bone marrow mesenchymal stem cells remains unclear.OBJECTIVE:To investigate the effect of carboxymethyl chitosan thermosensitive hydrogel on the differentiation of bone marrow mesenchymal stem cells into neurons and the possible mechanism.METHODS:Passage 3 bone marrow mesenchymal stem cells from rats were selected and cultured in carboxymethyl chitosan thermosensitive hydrogel extracts in different concentrations (0, 50, 100, 150, 200, 500 g/L). Control cells were cultured in culture medium with no addition of carboxymethyl chitosan thermosensitive hydrogel extracts. MTT assay was performed to investigate the effects of different concentrations of carboxymethyl chitosan thermosensitive hydrogel extracts on bone marrow mesenchymal stem cell proliferation. Western blot assay was used to explore the effect of 150 g/L carboxymethyl chitosan thermosensitive hydrogel extracts on the expression of neuron-specific enolase, Nestin, Vimentin, NF-M, microtubule associated protein 2, glial fibrillary acidic protein, β3-tubulin, Notch1 and Jag1 protein.RESULTS AND CONCLUSION:MTT assay showed that carboxymethyl chitosan thermosensitive hydrogel promoted the cell proliferation, and the proliferation rate reached the peak at the concentration of 150 g/L. Western blot assay showed that the cells induced by 150 g/L carboxymethyl chitosan thermosensitive hydrogel extract had significant increases in neuron-specific enolase, Nestin, Vimentin, NF-M, microtubule associated protein 2, glial fibrillary acidic protein, and β3-tubulin protein expression, and obvious decreases in Notch1 and Jag1 protein expression in comparison with the control group. These results indicate that the carboxymethyl chitosan thermosensitive hydrogel induces rat bone marrow mesenchymal stem cells to differentiate toward neurons, and suppresses the activity of Notch signal pathway in the process of differentiation.
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BACKGROUND:Choosing an effective means to label and trace the distribution, differentiation and migration of celsin vivo help to further explore the specific mechanism of cels that exert a therapeutic effect. OBJECTIVE:To understand the migration and localization of BrdU-labeled human umbilical cord mesenchymal stem cels in brain injury model rats. METHODS:Human umbilical cord blood samples were obtained, and the isolation of human umbilical cord mesenchymal stem cels was carried out. The primary and passage culture were performed. The phenotype of cels was detected by flow cytometry. Passage 3 human umbilical cord mesenchymal stem cels were labeled using BrdU, and the cel proliferation was detected using MTT method. BrdU-labeled cels were injected into brain injury ratsvia the tail vein. At 14 days after transplantation, brain tissues in the injury region were cut into sections and the migration and location of the umbilical cord mesenchymal stem cels were observed under inverted fluorescence microscope. RESULTS AND CONCLUSION: Cel surface specific markers CD45 and CD34 were detected by flow cytometry, but the cels could not express CD44, CD105 and CD29. Based on the cel growth curve, the cels came into a conditioning period at 1-3 days of seeding and came into a logarithmic phase at 3-5 days. BrdU-positive cels were visible at the injury region after 14 days, indicating that in the rats, transplanted human umbilical cord mesenchymal stem cels migrated from the peripheral blood to the site of brain injury to achieve the effective repair of injured parts. Cite this article:Liu HL, Liu ZJ, Chen XB, Hu WZ, Ding BQ. Migration and localization of umbilical cord mesenchymal stem cels implanted into brain injury model rats. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):31-35.
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BACKGROUND:Bone marrow stromal cel s can differentiate into nerve cel s to promote nerve tissue repair, but the exact mechanism has not been ful y elucidated. OBJECTIVE:To explore the influence of adenovirus-mediatedβnerve growth factor transfection on bone marrow stromal stem cel transplantation fighting against brain injury in rats. METHODS:(1) Rat bone marrow stromal stem cel s were cultured in vitro, transfected with the adenovirus-mediatedβnerve growth factor and directional y induced usingβ-mercaptoethanol. (2) A total of 210 Sprague-Dawley rats were randomized into induction+tranfection group, induction+non-transfection group, induction+medium group, model group, and sham group (n=42 per group). Rat skul injury models were made, and given corresponding treatments at different time points (12, 24, 36, 48, 72 hours). Neurological function of rats was evaluated based on neurological severity scores on the day that the rats were given transplantation, and 1, 2, 3, 4 weeks after transplantation. (3) Another 75 Sprague-Dawley rats were also divided into five groups (n=15 per group) as above, fol owed by model establishment and corresponding treatments at 24 hours after modeling. Neurological severity scores were recorded at the same day, 1, 2, 3, 4 weeks after transplantation. Five rats from each group were sacrificed to detect levels of malondialdehyde and superoxide dismutase in the rat brain at the same day, 2 and 4 weeks after transplantation, respectively. RESULTS AND CONCLUSION:If the cel s were transplanted within 48 hours after modeling, the neurological severity scores in the induction+transfection group decreased significantly compared with the induction+non-transfection group and model group at 1 and 2 weeks after transplantation (P<0.05). If the cel s were transplanted at different time, the neurological severity scores in the induction+transfection group were decreased significantly compared with the induction+non-transfection group and model group at 3 and 4 weeks after transplantation (P<0.05). If the cel s were transplanted within 24 hours after modeling, the neurological severity scores in the induction+transfection group decreased significantly compared with the model group at 1 week after transplantation (P<0.05), and the neurological severity scores in the induction+transfection group and induction+non-transfection group both were significantly lower than those in the model group (P<0.05). Two weeks after cel transplantation, the level of superoxide dismutase was significantly higher in the induction+transfection group than the induction+medium group and model group (P<0.05), but the level of malondialdehyde was significantly lower (P<0.05). Al these findings indicate that adenovirus-mediatedβnerve growth factor transfer plays a certain neuroprotective role in bone marrow stromal stem cel transplantation for brain injury in rats.
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According to the problem existing in PBL teaching of biochemistry, we reformed teaching approach, adjusted curriculum content, reassembled teaching material system, emphasized clinical practice, strengthened the experiment technique, expanded open class, created essential condition, built independent learning atmosphere actively, which promoted teaching quality enormously and demonstrated great advantage of PBL teaching method. Meanwhile, it made the curriculum reform of biochemistry carried out deeply.
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Objective To investigate the effects and significance of neuronavigation and electrocorticography monitoring in resection of eloquent brain glioma. Methods Thirty-six cases with intracranial tumors accepted microneurosurgery resection under neuronavigation and electrocorticography monitoring. The clinical data and postoperative outcome were analyzed. Results The mean registration error was (2.0 ±0. 5)mm in all operations and all skin flaps and bone windows designed by neuronavigation could fit the operation demands. Total resectin of the tumor was achieved in 31 cases and subtotal resection in 5 cases. Neurological symptoms improved and no severe complications or death happened in all patients. Conclusion Neuronavigation combined with electrocorticography monitoring can accurately locate the eloquent glioma and retrieve the brain shift. This method is a real-time technique and has functional test ability. It can improve the total removal rate and decrease the mortality and disabled rate.
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Objective To improve the preoperative diagnosis accuracy of cerebellopontine angle tumors through analyzing MRI findings. Methods Ninety-six cases with cerebellopontine angle tumors, confirmed by pathology and surgery, were collected and underwent MRI scan plus enhanced MRI. Among the 96 capes, we observed acoustic neurinoma in 55 cases, meningioma in 20 cases, cholesteatoma in 9 cases, trigeminal neuroma in 7 cases,cavernous hemangioma in 3 cases,arachnoid cyst in 2 cases. The MRI characteristics of all cases were analyzed retrospectively. Results The chief type of tumor happened in the cerebellopontine angle zone was acoustic neurinoma,followed in order by meningioma,cholesteatoma,trigeminal neuroma,cavernous hemangioma and arachnoid cyst. The accuracy of preoperative localization and qualitative diagnosis were 100% and 94.7%respectively.Conclusion MRI has a high value in the diagnosis and differential diagnosis of cerebellopontine angle tumors,which can be used as a preferred preoperative examination method in cerebellopontine angle tumors.
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Objective To investigate the effects of lymphangiogenesis on cancer growth and metastasis.Methods Human lymphatic endothelial cells(HLyECs)were isolated and purified from human lymph node by magnetic beads coated with antibody against human VEGFR3.Human breast cancer cell line(MDA-MB-435)or human osteosarcoma cell line(MG-63)was inoculated alone or co-inoculated with HLyECs subcutaneously into nude mice.The weight of tumor and lung surface metastasis were detected;peri-tumor lymphatic vessels were shown by Evans blue,intra-tumor lymphatic vessels were shown by immunohistochemistry for human and mouse PDPN.Conditioned medium of tumor on proliferation of HLyECs was evaluated by MTT assay.Results Compared with inoculating tumor cells alone,the growth and metastasis of MDA-MB-435 were promoted by co-inoculating HLyECs.The peritumoral and intra-tumoral lymphatic vessel density of MDA-MB-435 were increased by co-inoculating HLyECs.Both hPDPN-and mPDPN-positive lymphatic vessels were found.But co-inoculating HLyECs had no effect on lymphatic vessels density of MG-63.Neither intra-nor peri-tumor lymphatic vessels were found.The proliferation of HLyECs was increased by conditioned medium of MDA-MB-435 but not by MG-63.Conclusion Growth and lymphangiogenesis of breast cancer can be enhanced by co-inoculating lymphatic endothelial cells.
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Epigenetic modifications, including DNA methylation, histone modifications, RNA epigenetics and nucleosome remodeling, are important for gene transcription, but such modifications do not change the coding sequence of the gene. Although these events are heritable, they are potentially reversible, thus opening up opportunities for the therapy of cancer. So epigenetic modifications have been thrusted into the forefront of new drug discoveries. Current knowledge suggests that the agents that intervene epigenetics by "turning back on" silenced genes may represent a significant advancement in treating many forms of cancer. In this review, we summarized the research progress of epigenetic gene regulation and the proteins which could read epigenetic codes. And the relationship between epigenetics and cancer was discussed comprehensively.
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Humanos , Metilação de DNA , Epigênese Genética , Genética , Regulação da Expressão Gênica , Genética , Histonas , Metabolismo , Neoplasias , Genética , TerapêuticaRESUMO
DNA methylation is crucial for mammalian development, and DNA methylation is always in the dynamic status during preimplantation mouse embryos development. The effects of 5-AZA-CdR on the development of preimplantation mouse embryos were evaluated. Preimplantation mouse embryos created by in vitro fertilization were cultured continuously in 5-AZA-CdR (0.2, 1.0, or 5.0 μmol/L). Fertilized oocytes exposed to CZB containing 5-AZA-CdR at the pronuclear stage were unable to form morulae (0.2 and 1.0 μmol/L) or 4-cell embryos (5.0 μmol/L), while 2-cell stage embryos exposed to 5-AZA-CdR developed into uncompacted 8-cell (0.2 and 1.0 μmol/L) or 3/4-cell (5.0 μmol/L) stage embryos. The rate of morula formation was significantly lower in 4-cell embryos cultured in 5-AZA-CdR (1.0 or 5.0 μmol/L) than that in control embryos (P < 0.05). These data indicate that 5-AZA-CdR inhibits the development of mouse preimplantation embryos. Apoptosis, DNA methylation, and transcriptional activity were analyzed to determine the reason for these developmental defects. An aunexin V-PI assay revealed that high doses of 5-AZA-CdR led to apoptosis. Compared to the controls, DNA methylation was significantly reduced in uncompacted 8-cell embryos and morulae (p < 0.05) in a dose- dependent manner, whereas no significant change was detected in 2-or 4-cell embryos (P > 0.05). The observed changes in transcriptional activity, determined by measuring the incorporation of BrUTP, were similar to the observed alterations in DNA methylation. Therefore, the developmental defects induced by 5-AZA-CdR appear to bc mediated by alterations in DNA methylation and transcriptional activity in preimplantation mouse embryos.
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Objective To investigate and analyse the effect of PBL teaching method on the clincal medical students of different level,furthermore,to provide an effective guide for biochemistry teaching method reform.Methods Comparing the results of traditional teaching method and PBL teaching method in biochemistry on the clinical medical students of different level,and analyzing the applicable of PBL to various students.Results The seven-year students had a significant higher score in sum record by using PBL teaching method(P
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(0.05)) CONCLUSION: CL1 has inhibitory effect on growth of SGC-7901 cells in vitro,but does not affect proliferation of CD34~+ and differentiation of CD34~+ haematopoietic stem/progenitor cells to granulocyte and monocytes.This demonstrates that CL1 has anticancer effect,but does not affect the proliferation and differentiation of haematopoietic stem/progenitor cell to granulocytes and monocytes.It may not cause obvious haematopoietic depression.
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To adapt the requirements of cultivating high-quality medical talents,we implemented autonomous management model on selecting courses,research subjects,classic discussion,experiments and practice. This reformation is effective. One step further,we illustrated its significance from medical view: doctors,patients and diseases. Meanwhile,it was pointed out that the autonomous management is an inevitable trend for management development in the future.