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Bupleuri Radix is commonly used in the traditional Chinese medicine, and saikosaponins are the important active ingredients. In this study, we first established a relative quantitative method for 25 saikosaponins using ultra high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-QTrap-MS) in the scheduled multiple reaction monitoring (sMRM) mode. The established method showed good intra-day and intra-day precision, linearity, repeatability and stability. Then the method was applied to compare 37 batches of Bupleuri Radix from different planting areas. The results showed that there was no significant difference in the saikosaponins composition of Bupleuri Radix from different planting areas in Shanxi Province, which indicating that Bupleuri Radix is well adapted to the environment, so it is suitable for widely planting. However, Bupleuri Radix harvested at spring and autumn were differed from those harvested at summer, which indicated that the traditional harvesting experience was reasonable. Correlation analysis showed that saikosaponins a and d were positively correlated with some saponins, and 4 saponins (such as clinoposaponin XII) showed bigger content variation were identified by coefficient of variation analysis. The LC-MS based pseudotargeted metabonomic method established in this study can be applied to the comprehensive detection of saikosaponins, which providing new method for the quality evaluation of Bupleuri Radix.
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OBJECTIVE@#To investigate the effect of Rheb1 in the development of mouse megakaryocyte-erythroid progenitor cells and its related mechanism.@*METHODS@#Rheb1 was specifically knocked-out in the hematopoietic system of Vav1-Cre;Rheb1fl/fl mice(Rheb1Δ/Δ mice). Flow cytometry was used to detect the percentage of red blood cells in peripheral blood and erythroid cells in bone marrow in Vav1-Cre;Rheb1fl/fl mice and control mice. The CFC assay was used to detect the differentiation ability of Rheb1 KO megakaryocyte-erythroid progenitor cells and control cells. Real-time fluorescence quantification PCR was used to detect the relative expression of PU.1,GATA-1,GATA-2,CEBPα and CEBPβ of Rheb1 KO megakaryocyte-erythroid progenitor cells and control cells. Rapamycin was added to the culture medium, and it was used to detect the changes in cloning ability of megakaryocyte-erythroid progenitor cells from wild-type mice in vitro.@*RESULTS@#After Rheb1 was knocked out, the development and stress response ability of megakaryocyte-erythroid progenitor cells in mice were weaken and the differentiation ability of megakaryocyte-erythroid progenitor cells in vitro was weaken. Moreover, the expression of GATA-1 of megakaryocyte-erythroid progenitor cells was decreased. Further, rapamycin could inhibit the differentiative capacity of megakaryocyte-erythroid progenitor cells in vitro.@*CONCLUSION@#Rheb1 can regulate the development of megakaryocyte-erythroid progenitor cells probably through the mTOR signaling pathway in mice.
Subject(s)
Animals , Mice , Cell Differentiation , Erythrocytes , Flow Cytometry , Megakaryocyte-Erythroid Progenitor Cells , Megakaryocytes , Signal TransductionABSTRACT
With the acceleration of the aging society, neurodegenerative diseases, such as Alzheimer's disease (AD) and Parkinson's disease (PD), have become a rapidly growing global health crisis. Recent studies have indicated that microglia-neuron interactions are critical for maintaining homeostasis of the central nervous system. Genome-Wide Association Studies and brain imaging studies have suggested that microglia are activated in early stage of neurodegenerative diseases. Microglia are specialized phagocytes in the brain. The discovery of a new phagocytic pathway, trogocytosis, suggests that there is a close interaction between microglia and surviving neurons. In this review, we summarize the important roles of microglia in neurodegenerative diseases, and further analyze the functions and molecular mechanisms of microglia phagocytosis and trogocytosis.
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Humans , Alzheimer Disease , Genome-Wide Association Study , Microglia/metabolism , Neurodegenerative Diseases , Phagocytosis/physiologyABSTRACT
OBJECTIVE@#To study the effect and mechanism of Huayu Wan (, HYW) in combination of chemotherapy of tumor treatment.@*METHODS@#HYW serum was added in Lewis cells to assess its impact on fluorescent doxorubicin delivery in vitro. Then, Lewis tumor cells was implanted in C57BL/6 mice via xenograft transplantation. Tumor growth was measured and signal intensity corresponding to blood flow was assessed by laser doppler perfusion imaging (LDPI). Finally, the effect of HYW on the effificacy of doxorubicin was studied.@*RESULTS@#HYW can improve the transfer of fluorescent doxorubicin into cells. The blood flow signal in the tumor tissues of the HYW group was higher than that of the control group (P<0.01). Furthermore, HYW improved drug delivery of doxorubicin to tumor tissues, and this activity was associated with HYW-induced microvascular proliferation (P<0.01).@*CONCLUSIONS@#HYW can promote microangiogenesis and increase blood supply in tumor tissues, which in turn may increase the risk of metastasis. At the same time, HYW increases drug delivery and improves the effificacy of chemotherapy drugs through vascular proliferation. Therefore, rational judgment must be exercised when considering applying HYW to an antitumor regimen.
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Objective:Death causes and life reduction of malignant tumors in the residents of Zhuanqiao Town in Minhang District from 2013 to 2017 were analyzed to provide scientific evidence for the strategies on comprehensive prevention and control of cancer. Methods:The data of death causes of malignant tumors in the residents of Zhuanqiao Town were collected and analyzed. The mortality rate， annual percent change （APC）， composition ratio， potential years of life lost （PYLL）， potential years of life lost rate （PYLLR） and average years of life lost （AYLL） of the registered population were analyzed. Results:The standardized mortality rate of malignant tumors in the residents of Zhuanqiao Town from 2013 to 2017 was 128.05/105， and the rate was higher in males than that in females. The top four cancers regarding PYLL were lung cancer， liver cancer， stomach cancer and colorectal cancer， which were roughly the same order as the top four regarding the mortality. This indicates that these four cancers had a greater impact on residents. Lung cancer had a greater impact on female life expectancy. PYLL and SPYLL ranked the first in liver cancer in males and thus had a greater impact on the males. Breast cancer was one of the most important malignant tumors in causing early death of women. Conclusion:Malignant tumor has become an important public health problem endangering the health of residents. The focus of future work in the town remains to improve public awareness of carcinogenic risk factors， actively carry out health education， lifestyle intervention and early screening in order to reduce cancer risk， alleviate cancer burden and improve the life expectancy of residents.
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OBJECTIVE@#To investigate the expression level of miR-181b in CD19+ B lymphocytes of patients with chronic lymphocytic leukemia (CLL), to analyze the relationship between its expression and the prognosis of CLL patients, and to predict the potential target gene of miR-181b in CLL by using bioinformatics.@*METHODS@#Eight-four patients with CLL treated in People's Hospital of Xinjiang Uygur Autonomous Region from June 2013 to June 2018 were selected. and 20 healthy people were selected as control group. RNA was extracted from CD19+B lymphocytes of peripheral blood by magnetic bead sorting, the expression level of miR-181b was detected, and it's expression differences in different IPI groups were analyzed. The correlation between the expression level of miR-181b and PFS of CLL patients also was analyzed. miR-181b target genes were predicted by online database and literatures, and gene annotation analysis and relevant signal pathway analysis were performed for candidate target genes.@*RESULTS@#The expression level of miR-181b in CLL patients was significantly lower than that in control group (P＜0.01); The expression level of miR-181b in the low-risk group was higher than that in high-risk group and extremely high-risk group (P＜0.05), but there was no statistical difference between low-risk group and medium-risk group (P=1.00). The expression level of miR-181b in medium-risk group was higher than that in high-risk group and extremely high-risk group (P＜0.05), but there was no difference between high-risk group and extremely high-risk group (P=1.00). ROC curve results showed that the area under the curve (AUC) was 0.792 (P＜0.01).When the expression level of miR-181b was at the threshold value of 0.279, it showed a better sensitivity (62.9%) and specificity (91.8%). Survival analysis results suggested that compared with the high expression group, the miR-181b low expression group had poor PFS (log rank: P=0.047). Prediction of miR-181b by using the starBase, targetscan and picTar database and its combination with literature reports indicated that CARD11, ZFP36L1, RUNX1, NR4A3, ATP1B1, PUM1 and PLAG1 related with blood diseases, and up-regulated CARD11 and ZFP36L1 participated in lymphoid tumor formation by promoting cell proliferation and inhibiting cell aging.@*CONCLUSION@#The expression level of miR-181b in CLL group are significantly lower than that in the controls group, and the low expression of miR-181b relates with poor prognosis of CLL patients. Through bioinformatics prediction and combined with literature reports, it is speculated that CARD11 and ZFP36L1 as target genes of miR-181b may be participated in the occurrence and development of CLL. Further experiments are needed to verify this result.
Subject(s)
Humans , Apoptosis Regulatory Proteins , Cell Proliferation , Leukemia, Lymphocytic, Chronic, B-Cell , Genetics , MicroRNAs , PrognosisABSTRACT
OBJECTIVE@#To explore the possible underlying mechanism by investigating the effect of electroacupuncture (EA) treatment on the primary motor cortex and striatum in a unilateral 6-hydroxydopamine (6-OHDA) induced rat Parkinson's disease (PD) model.@*METHODS@#Male Sprague-Dawley rats were randomly divided into sham group (n=16), model group (n=14), and EA group (n=14). EA stimulation at Dazhui (GV 14) and Baihui (GV20) was applied to PD rats in the EA group for 4 weeks. Behavioral tests were conducted to evaluate the effectiveness of EA treatment. Metabolites were detected by 7.0 T proton nuclear magnetic resonance.@*RESULTS@#Following 4 weeks of EA treatment in PD model rats, the abnormal behavioral impairment induced by 6-OHDA was alleviated. In monitoring changes in metabolic activity, ratios of myoinositol/creatine (Cr) and N-acetyl aspartate (NAA)/Cr in the primary motor cortex were significantly lower at the injected side than the non-injected side in PD rats (P=0.024 and 0.020). The ratios of glutamate + glutamine (Glx)/Cr and NAA/Cr in the striatum were higher and lower, respectively, at the injected side than the non-injected side (P=0.046 and 0.008). EA treatment restored the balance of metabolic activity in the primary motor cortex and striatum. In addition, the taurine/Cr ratio and Glx/Cr ratio were elevated in the striatum of PD model rats compared to sham-lesioned rats (P=0.026 and 0.000). EA treatment alleviated the excessive glutamatergic transmission by down-regulating the striatal Glx/Cr ratio (P=0.001). The Glx/Cr ratio was negatively correlated with floor plane spontaneous locomotion in PD rats (P=0.027 and P=0.0007).@*CONCLUSIONS@#EA treatment is able to normalize the metabolic balance in the primary motor cortex and striatum of PD rats, which may contribute to its therapeutic effect on motor deficits. The striatal Glx/Cr ratio may serve as a potential indicator of PD and a therapeutic target of EA treatment.
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Objective:To evaluate and compare the identification of several DNA barcoding candidate sequences on Illicium difengpi and its fake I. jiadifengpi. Method:Samples from different origins of I. difengpi and I. jiadifengpi, were collect extraction of total DNA,nuclear gene ITS2 sequence,chloroplast rbcL,matK gene sequence were selected for PCR amplification,product purification and sequencing,and CondonCode Aligner V3.7.1 was used to proofread stitching. Result:PCR amplification and sequencing of rbcL sequences of I. difengpi and I. jiadifengpi were not satisfactory. It is assumed that their rbcL sequences were too long with slow evolution,which was unsuitable for interbreeding. The success rate of matK sequencing of I. difengpi and I. jiadifengpi was 0 and 76.8%,which may be because primer standards were different for matK sequences of different groups. The results of PCR amplification and sequencing of ITS2 on I. difengpi and I. jiadifengpi were successful,with the success rate of sequencing was 89.3% and 91.2%. Analysis sequencing results, the total length of ITS2 sequences was 268 bases,and there were 2 variation sites of I. difengpi. The total length of ITS2 sequences was 430 bases,and there were 4 or 3 variation sites of I. jiadifengpi. It shows that ITS2 sequences of I. difengpi and I. jiadifengpi were short and has obvious variability and can be amplify,that ITS2 sequence was better than rbcL and matK sequence in molecular identification of I. difengpi and I. jiadifengpi. Conclusion:DNA barcoding based on ITS2 sequence was a powerful and efficient tool for identification of I. difengpi and its fake I. jiadifengpi.
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Metabolic remodeling, a well-recognized hallmark of cancer, provides biomass and energy to support the growing demand of unrestricted growth of cancer cells. In addition to metabolic supplies, the altered cell metabolism often results in the intracellular accumulation of particular metabolites. Recently, it is increasingly revealed these metabolites may exhibit metabolism-independent roles as signaling molecules, which triggers oncogenic signaling via various mechanisms including competitive inhibition, protein post-translational modifications and direct protein binding. These insights provide a new perspective of metabolic remodeling in cancer progression. This review summarized the recent advancement in the understanding of metabolites as signaling molecules independent rewriting of metabolic pathways. By summarizing these progresses, this review hopes to provide a better understanding of tumor metabolic disturbance and to contribute to the discovery of novel potential therapeutic targets.
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To accurately discriminate Stellariae Radix from its adulterants, four leading candidate DNA barcoding markers were evaluated. Sixty samples including Stellariae Radix and its adulterants have been newly collected and their total genomic DNA was extracted. Four DNA barcoding markers ITS, rbcL, psbA-trnH and matK were amplified and sequenced. Their sequence characteristic analyses, Kimura-2-parameter (K2P) distance calculation and Neighbor-joining (NJ) phylogenetic tree constructions were accomplished using the MEGA 7.0 software. DNA Barcoding gaps of the four DNA barcoding markers were estimated by the distributions of inter- and intra-sequence specific variations. Species identification efficiency was calculated using the BLAST method. The results showed that ITS had the highest (95.2%) while matK demonstrated the lowest (75%) PCR and sequencing efficiency. The length range of the four markers were in the ranger of 211-797 bp, and the G+C content of ITS was highest (54.35%). The identification efficiency of matK and ITS was 92% and 90% respectively. Barcoding gap could be found in ITS sequences. The NJ phylogenetic tree constructed using ITS sequences showed that samples of Stellariae Radix were separately formed into one clade, and samples of adulterants like Stellaria bistyla were clearly belong to different branches from Stellariae Radix, whereas NJ trees constructed using psbA-trnH, rbcL and matK could not differentiate Stellariae Radix from its adulterants. Therefore, ITS regions as DNA barcodes can stably and accurately distinguished Stellariae Radix from its adulterants, and provide a new technique for modern identification of Stellariae Radix.
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Objective To investigate the prevalence and influencing factors of mild cognitive impairment (MCI) among elderly people in communities of Nanning, so as to provide scientific basis for the formulation of MCI early intervention measures. Methods A total of sample of 3 000 elderly people aged 60 or above living in Nanning area were sampled out using the method of cluster random sampling from three communities in Nanning. The Beijing version of the montreal cognitive assessment (MOCA-BJ) was administered by face to face interview. Results 833 MCI patients were detected. The standardized prevalence rate of MCI was 27.27% by the age composition of the population in Nanning in 2010. Single factor analysis showed that there were significant differences(all P<0.05)in the prevalence of MCI in different groups by age, education, occupation, exercise time, sleep time, number of reading, community activities and housework. Multivariate analysis showed that old age, primary education level, occupation of farmer/migrant worker and enterprise personnel/worker were the risk factors of MCI, adequate sleep time, moderate exercise time and reading times were the protective factors of MCI (all P<0.05). Conclusions The prevalence rate of MCI is high among the elderly in Nanning. The elderly with advanced age, primary school education, occupation as farmer/peasant-worker and enterprise personnel/workerare at high risk of developing MCI. Effective measures should be taken as soon as possible to prevent the occurrence and development of MCI.
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Chimonanthus plants widely distributed in southern area of China, which have a long history of edibles and medicine. Phytochemical investigations have shown that Chimonanthus produced 143 non-volatile constituents, including alkaloids, flavonoids, terpenoids, coumarins and others, which exhibit significant anti-oxidant, anti-bacterial, anti-cancer, anti-inflammatory, antihyperglycemic, antihyperlipidemic and other biological activities. On the basis of systematic reviewing of literatures, this article overviews the non-volatile constituents and pharmacology of Chimonanthus from domestic and foreign over the last 30 years (until June 2018), and may provide a useful reference for the further development of Chimonanthus.
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Animals , Humans , Calycanthaceae , Chemistry , Drugs, Chinese Herbal , Chemistry , Pharmacology , Therapeutic Uses , Toxicity , Medicine, Chinese Traditional , Phytochemicals , Chemistry , Pharmacology , Therapeutic Uses , Toxicity , PhytotherapyABSTRACT
After the advantages and processes of medical library in development of think tank were described with the practice of Capital Medical University Library as an example,the role of academic medical libraries in providing reference for decision-making for subject construction in colleges and universities was studied on the basis of subject bibliometric evaluation and keeping abreast of subject frontiers in their think tank development-oriented subject service.
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<p><b>OBJECTIVE</b>The prevalence rate, distribution, and reasons of immature permanent-tooth trauma in Xi'an were investigated and described, and a scientific basis was provided for the decision-making of health-administration departments.</p><p><b>METHODS</b>Through cluster and simple random sampling survey, 4 013 pupils in schools from nine districts and four counties in Xi'an were sampled. All pupils and their parents were requested to complete a questionnaire. Oral examinations were conducted for children who had immature permanent teeth trauma. The number of trauma teeth and teeth were recorded. Survey data were analyzed statistically.</p><p><b>RESULTS</b>The prevalence rate of immature permanent dental trauma was 10.5% in Xi'an. No significant differences were observed between districts and counties (P>0.05). The peak age of permanent dental trauma was at 7-9 years old, and the most was at 8 years old (31.5%). The first reason of trauma was falling down (50.9%), and the second reason was crash (36.0%). The most common trauma teeth were maxillary incisors (75.4%).</p><p><b>CONCLUSIONS</b>The prevalence rate of immature permanent teeth trauma in Xi'an was in the middle of all international levels. Children, who are vulnerable to dental trauma at their age, should be provided with appropriate prevention measures to reduce the incidence of dental trauma.</p>
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<p><b>OBJECTIVE</b>This study aims to investigate the clinical visiting and prognosis of schoolchildren in Xi'an after immature permanent tooth trauma and explore the prognostic factors associated with this type of trauma.</p><p><b>METHODS</b>Through cluster and simple random sampling surveys, 4 013 pupils in schools from nine districts and four counties in Xi'an were sampled, respectively. All pupils and their parents were requested to complete a questionnaire, and children who had immature permanent tooth trauma answered a separate questionnaire and underwent oral examinations. The data of the survey were analyzed statistically.</p><p><b>RESULTS</b>The amount of valid questionnaire was 3 641. Clinical visiting rate related to immature permanent tooth trauma of Xi'an schoolchildren was low (38.2%). Gender and trauma type were the factors related to clinical visiting for dental trauma concerns. The incidence of poor prognosis was 29.7%. The incidence of poor prognosis of patients with clinical visiting (35.4%) was higher than that of patients without clinical visiting (20.6%).</p><p><b>CONCLUSIONS</b>Clinical visiting rate related to immature permanent tooth trauma in Xi'an schoolchildren is extremely low, and incidence of poor prognosis is high.</p>
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<p><b>OBJECTIVE</b>To assess the effectiveness of Yishen Jiangu Granules (, YSJGG) on aromatase inhibitor-associated musculoskeletal symptoms (AIMSS).</p><p><b>METHODS</b>A single-arm, open-label study was conducted in 34 postmenopausal women with breast cancer who experienced AIMSS. Patients were treated with YSJGG for 12 weeks (12.4 g orally twice daily). The primary outcome was a change in the mean worst pain score of Brief Pain Inventory-Short Form (BPI-SF) over 12 weeks, and the second outcomes included changes in pain severity and pain-related interference of BPI-SF and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Modified Score for the Assessment of Chronic Rheumatoid Affections of the Hands (M-SACRAH), the Functional Assessment of Cancer Therapy-Breast (FACT-B), bone mineral density (BMD) and blood indices such as calcium (Ca), phosphate (P), and alkaline phosphatase (ALP).</p><p><b>RESULTS</b>Of 37 women recruited, 30 initiated the therapy and 24 were evaluable at 12 weeks. The primary outcome (BPI-SF worst pain scores) achieved a 2.17-point reduction compared with baseline (5.75±1.87 vs 3.58±2.15, P<0.01). There were reductions in pain severity (decreased 1.65, P<0.01) and pain-related interference (decreased 2.55, P<0.01). The changes in WOMAC and M-SACRAH scores were similar to BPI-SF (P<0.05). In the FACT-B, only physical well-being and functional well-being were improved compared with baseline (P<0.05). No clinical differences were found in BMD, Ca, P and ALP.</p><p><b>CONCLUSION</b>YSJGG is an effective and well-tolerated agent to reduce AIMSS.</p>
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<p><b>OBJECTIVE</b>To evaluate and compare the clinical efficacy and safety of recombinant human thrombopoietin(rhTPO) and recombinant human interleukin11(rhIL-11) for the treatment of chemotherapy-induced thrombocytopenia in adult acute myeloid leukaemia patients.</p><p><b>METHODS</b>Total of 96 adult acute myeloid leukaemia patients were divided into 3 groups according to randomized controlled method: rhTPO group, rhIL-11 group and control group, 32 cases in each group. The patients in rhTPO group and rhIL-11 received rhTPO of 15000 IU/d and rhIL-11 of 1.5 mg/d, respectively after the standard combined chemotherapy within 24 hours, and patients in control group, received nothing drugs to promote thrombocyte recovery. And rhTPO and rhIL-11 should be stopped when the Plt≥100× 10/L. After chemotherapy, the platelet recovery degree, duration of Plt<50× 10/L, ≥50× 10/L and ≥100× 10/L, the count of infusion thrombocytes, and incidence of adverse reactions all were compared.</p><p><b>RESULTS</b>The duration of Plt<50× 10/L was obviously less than that in control group(P<0.01). The duration of rhIL-11 was less than that in control group, but there was no statistical significance(P>0.05). As compared with that in control group, the Plt count in rhTPO and rhIL-11 groups can faster increase to Plt≥50× 10/L (P<0.01, P<0.05), among them the Plt count in rhTPO group faster increase, but there was no statistical signiticance. As compared with that in control group, the Plt count in rhTPO group and rhIL-11 group can increase to Plt≥100× 10/L (P<0.01), the Plt count in rhTPO group was more obviously increase than that in rhIL-11 group(P<0.05). The count of infusion Plt in rhTPO and rhIL-11 groups was lese than that in control group(P<0.01, P<0.05), and the count of infusion Plt in rhTPO group was less than that in rhIL-11 group(P<0.05). After using rhTPO and rhIL-11, the adverse reactions, such as low fever, induration of injection site, athralgia, nausea and vomiting occured in rhTPO group and rhIL-11 group, but all can be tolerated.</p><p><b>CONCLUSION</b>Both rhTPO and rhIL-11 can reduce the duration of thrombocytopenia and the amount of infused thrombocyte, promote platelet recovery in the patients with acute myeloid leukaemia after chemotherapy, to decreae the risk of bleeding, and reduce incidence of adverse reactions, both of them can be tolerated by patients, and rhTPO is more advantage than rhIL-11, worthy of clinical popularization and application.</p>
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Objective The purpose of this study was to analyze the burden of stroke in Fenghua City by disability-adjusted life-years (DALYs).Methods We used data of stroke incidence,prevalence and mortality in Fenghua City in 2013 and selected DISMODII to calculate DALYs.Results There were 1 895 cases of stroke reported in 2013,and the reported incidence rate was 392.03/100 000(standard incidence rate was 201.14/100 000).The reported incidence rate among male 436.33/100 000 (standard incidence rate was 210.55/100 000 ),higher than that of female 346.48/100 000 (standard incidence rate was 189.25/100 000)(P<0.05).There were 751 cases of stroke death,and the mortality rate was 155.28/100 000(standard mortality rate was 66.53/100 000).The prevalence rate of stroke was 1 899.53/100 000, and people aged over 60 were accounted 85.53%.The overall disease burden of stroke was 7 566.03 DALYs,with the intensity of 15.64 DALYs per 1 000 population.Men had a higher DALYs and DALYs intensity than in women.The burden of residents aged over 60 was 84.75%.Years of life lost (YLL ) was the main fraction in DALYs,and its percentage was 76.75%.Conclusion The overall disease burden of stroke in Fenghua City was high,suggesting that the community should pay more attention to the men aged over 60 in order to reduce the burden of stroke.
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<p><b>OBJECTIVE</b>To explore the role of PDK1 in T-ALL development through establishing the Notch1-induced T-ALL mouse model by using Mx1-cre; LoxP system to knock-out PDK1.</p><p><b>METHODS</b>Cell cycle and apoptosis of leukemic cells were detected by flow cytometry, and relative expression of tumor-related genes and transcription factors of leukemic cells were determined by quantitative real-time PCR.</p><p><b>RESULTS</b>Notch1-induced T-ALL mouse model with inducible knock-out of PDK1 was established successfully. Compared to T-ALL control mouse model, PDK1 knock-out mice showed a significant longer survival time (P<0.01). There was no difference of cell cycle between control and PDK1 knock-out mice, and the apoptosis rate of leukemic cells in PDK1 knock-out mice was higher than that of control mice (P<0.001). PDK1 knock-out resulted in decreased expression of tumor-related genes and transcription factors, such as c-Myc and NF-κB (P<0.01), and increased expression level of P53 (P<0.01).</p><p><b>CONCLUSION</b>PDK1 knock-out can inhibit the development of T-ALL, and its mechanism may be the leukemia progression inhibited by regulating the apoptosis and expression of multiple related genes and transcription factors.</p>
Subject(s)
Animals , Mice , Apoptosis , Cell Cycle , Disease Models, Animal , Gene Expression Regulation, Leukemic , Mice, Knockout , NF-kappa B , Genetics , Metabolism , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma , Genetics , Protein Serine-Threonine Kinases , Genetics , Proto-Oncogene Proteins c-myc , Genetics , Metabolism , Real-Time Polymerase Chain Reaction , Receptor, Notch1 , Genetics , Tumor Suppressor Protein p53 , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of Rheb (mTOR activator) in AML development by measuring Rheb expression in bone marrow of adult AML patients and in AML cell line HL-60.</p><p><b>METHODS</b>Real-time PCR assay was used to measure the Rheb mRNA expression in 27 AML patients and 29 ITP patients as control. The relationship between Rheb mRNA expression and age, AML subtype, fusion gene, splenomegaly, hepatomegaly and survival of AML patients was analyzed and compared. In addition, HL-60 cell line over-expressing Rheb was established, and the HL-60 cells and HL-60 cells with overexpression of Rheb were treated with Ara-C of different concentrations, the proliferation level was detected by CCK-8 method, and the IC50 was calculated.</p><p><b>RESULTS</b>The mRNA level of Rheb in AML patients was similar to that in ITP patients (control). Interestingly, higher expression of Rheb was associated with better survival and was sensitive to Ara-C treatment. However, the expression level of Rheb was not associated with age, AML subtype, fusion gene, and hepatomegaly of patients. Lower expression level of Rheb was associated with splenomegaly. In vitro analysis of HL-60 line indicated that overexpression of Rheb could increased the cell sensitivity to Ara-C treatment (IC50=0.54 µmol/L) and caused HL-60 cell apoptosis.</p><p><b>CONCLUSION</b>The lower Rheb expression is a poor prognostic indicator for AML patients, which is associated with AML splenomegaly, the patients and HL-60 cells with low expression of Rheb are insensitive to Ara-C treatment.</p>