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ObjectiveTo investigate the mechanism of Renshen Guben oral liquids（RGOL） in treatment of mice with renal fibrosis based on metabolomics and network pharmacology. MethodC57BL/6 mice were randomly divided into control group， model group and RGOL group， 12 mice in each group. Except for the control group， mice in the other groups were induced into unilateral ureteral obstruction（UUO） model by UUO. After preparation of the model， an aqueous solution of 4.2 g·kg-1 extract powder was administered by gavage to RGOL group for 14 d， and an equal amount of distilled water was administered by gavage to the control and model groups. After the last administration on the 14th day， urine was collected and detected by ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry（UPLC-QQQ-MS/MS） with 0.1% formic acid aqueous solution as mobile phase A， and acetonitrile-isopropanol（70∶30） as mobile phase B for gradient elution（0-1 min， 5%B； 1-5 min， 5%-30%B； 5-9 min， 30%-50%B； 9-11 min， 50%-78%B； 11-13.5 min， 78%-95%B； 13.5-14 min， 95%-100%B； 14-16 min， 100%B； 16-16.1 min， 100%-5%B； 16.1-18 min， 5%B）， column temperature of 40 ℃， flow rate of 0.4 mL·min-1， electrospray ionization（ESI）， collection range of m/z 50-900. Through network pharmacology， the targets of components in RGOL and the targets of renal fibrosis were analyzed interactively， and the key components and key targets were screened by network topology analysis， and DAVID platform was used to predict the signaling pathways of RGOL for the treatment of renal fibrosis. ResultA total of 7 differential metabolites involving 8 metabolic pathways were identified in RGOL for the treatment of renal fibrosis. The network pharmacology revealed that 36 key components in RGOL were related to 7 differential metabolites， mainly ginsenosides， notoginsenosides and nucleotides. Based on the herbs-components-targets-pathways network， a total of 23 key targets related to the treatment of renal fibrosis by RGOL were highlighted， which together with the differential metabolites were involved in linoleic acid metabolism， arginine biosynthesis， tricarboxylic acid cycle（TCA）， arginine and proline metabolism and other pathways. ConclusionBased on metabolomics and network pharmacology， this study preliminarily identified 7 differential metabolites， 36 potential pharmacodynamic components and 23 key targets and 4 key pathways in RGOL for the treatment of renal fibrosis， providing an experimental basis for the clinical application and mechanism study of this preparation.
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Objective:To investigate the effect and mechanism of Myosin X on the radiosensitivity of non-small cell lung cancer (NSCLC) cell line H1975 in vitro. Methods:Western blot was applied to detect the expression level of Myosin X expression. The H1975 cell line with stable knockout of Myosin X (KO group) and infected with control virus (NC group) were constucted by using CRISPR/Cas9 technique. The knockout efficiency was validated. The radiosensitivity of two cell lines was measured by colony formation assay and single-hit multi-target model. γ-H 2AX focus formation test and RNA sequencing (RNAseq) analysis were employed to identify the regulatory mechanism of the radiosensitivity of lung cancer cell lines mediated by Myosin X. Results:The expression level of Myosin X in the H1975 cells was significantly up-regulated than those in other NSCLC cell lines (all P<0.01). The lentiviral vector of Myosin X sgRNA-Lenti-CRISPR v2 was successfully constructed. After the puromycin screening, H1975 cell lines with complete knockout of Myosin X and control cell lines (NC group) were obtained. Colony formation assay demonstrated that compared with the NC group, the radiosensitivity in the KO group was significantly higher (The D 0 value was decreased from 1.28 Gy to 1.03 Gy, SF 2 decreased from 0.29 to 0.21, and the sensitization ratio was 1.24). The γ-H 2AX focus formation test showed that the number of damage focus formed at 1 h and 6 h after irradiation in the KO group was significantly larger than that in the NC group ( P<0.05. RNAseq analysis indicated that the expression level of ISLR in the KO group was significantly down-regulated than that IN the NC group ( P<0.05). Conclusion:Knockout of Myosin X can increase the radiosensitivity of H1975 cells probably by interfering the repair of DNA double-strand damage and down-regulating the expression level of ISLR.
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OBJECTIVE：To ex plore the effects of solamargine on the growth and apoptosis of human hepatocarcinoma cells HepG2 and its underlying mechanism. METHODS ：The effects of 0（blank group ）-12 μmol/L solamargine treatment of 24，48 h on survival rate of HepG 2 cells were investigated. The effects of 0（blank group ），6 μmol/L solamargine treatment of 10 days on cell clone formation were also investigated. The effects of 0（blank group ），4，6，8 μmol/L solamargine for 24 h on the apoptotic rate of cells，mRNA expression of Bcl- 2，Bax and caspase- 3， protein expression of Bcl- 2 and cleaved caspase- 3 as well as ratio of p-AMPKα to AMPKα were all tested. The effects of AMPK inhibitor as compound C on the protein expression of AMPKα and Bcl- 2 in cells were investigated after treated with 6 μmol/L solamargine for 24 h. RESULTS ：Compared with 020-39318678。E-mail：wujingjing6028@gzucm.edu.cn blank group ，1-12 μ mol/L solamargine for 24，48 h could significantly decrease the survival rates of cells （P＜0.05）in a concentration-dependent manner ；IC50 of them were 8.310 and 7.996 μmol/L，respectively；the rate of cell clone formation was decreased significantly after treated with 6 μmol/L solamargine for 10 days（P＜0.05）. The apoptotic rate of HepG 2 cells，mRNA expression of Bax and caspase- 3，protein expression of cleaved caspase-3（except for 8 μmol/L）as well as ratio of p-AMPKα to AMPKα（except for 8 μmol/L）were all increased significantly after treated with 6，8 μmol/L solamargine（P＜0.05）；mRNA and protein expression of Bcl- 2 were decreased significantly （P＜ 0.05）；the changes of some indexes were in a concentration-dependent manner. The compound C could inhibit protein expression of AMPKα，and reverse the inhibitory effect of solamargine on Bcl- 2 protein. CONCLUSIONS ：Solamargine can inhibit the proliferation of HepG 2 cells and induce apoptosis ，the mechanism of which may be associated with activating AMPK signaling pathway.
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Over the past decade, traditional Chinese medicine (TCM) has widely embraced systems biology and its various data integration approaches to promote its modernization. Thus, integrative pharmacology-based traditional Chinese medicine (TCMIP) was proposed as a paradigm shift in TCM. This review focuses on the presentation of this novel concept and the main research contents, methodologies and applications of TCMIP. First, TCMIP is an interdisciplinary science that can establish qualitative and quantitative pharmacokinetics-pharmacodynamics (PK-PD) correlations through the integration of knowledge from multiple disciplines and techniques and from different PK-PD processes
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Objective:To explore the appropriate radiotherapy time and method in the treatment of patients with brain metastases (BM) due to from non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) mutation.Methods:Totally 69 EGFR-mutant NSCLC patients with BM treated in Zhongnan Hospital of Wuhan University from January 2014 to September 2018 were retrospectively reviewed. The patients were divided into two groups according to the time of brain radiotherapy, including the upfront radiotherapy group ( n=45) who received concurrent brain radiotherapy and EGFR-tyrosine kinase inhibitors(TKI)treatments and deferred radiotherapy group ( n=24) who received brain radiotherapy after intracranial progression during EGFR-TKI treatment. The upfront radiotherapy group was further divided into two groups, the group treated with WBRT concurrent with EGFR-TKI ( n=20) and the group treated with SRS concurrent with EGFR-TKI ( n=25). Overall survival (OS), progression-free survival (PFS) and intracranial progression-free survival (iPFS) time were evaluated. Results:The median OS of 69 patients was 31.2 months. For the upfront and deferred radiotherapy groups, the 1-, 2- year OS were 95%, 64% and 80%, 35%, the difference between the two groups was statistically significant. On subgroup analysis, the upfront WBRT, upfront SRS and deferred radiotherapy groups 1-, 2- year OS were 95%, 96%, 80% and 42%, 88%, 35%. Moreover, the upfront SRS group was associated with improved OS relative to the deferred radiotherapy group ( HR: 0.10, 95% CI: 0.23-0.46, P=0.003), but the upfront WBRT and deferred radiotherapy groups shared similar OS ( HR: 0.54, 95% CI: 0.21-1.32, P=0.180). There were no significant difference in iPFS and PFS between the upfront and deferred radiotherapy groups( P>0.05). Conclusions:Upfront brain radiotherapy prolonged the survival of BM patients metastasized from EGFR-mutant NSCLC. SRS concurrent with EGFR-TKI may be superior to WBRT concurrent with EGFR-TKI in the treatment of BM metastasized from EGFR-mutant NSCLC.
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Objective:To investigate the relationship between Helicobacter pylori(Hp)infection and carotid atherosclerosis(CAS)in the elderly, in order to provide an empirical basis for the prevention and treatment of cardiovascular and cerebrovascular diseases in the elderly.Methods:A total of 287 patients aged 60 years and over admitted to the First Affiliated Hospital of Guangxi Medical University, who underwent the 13C-urea breath test( 13C-UBT), carotid color and two-dimensional Doppler ultrasonography from October 2015 to January 2019, were retrospectively enrolled.Patients were divided into the Hp infection group(n=137)and the non-Hp infection group(n=150). Common high-risk pathogenic factors, blood biochemical indicators, carotid intima-media thickness(IMT)and detection rate of carotid plaque were compared between the two groups. Results:Common high-risk pathogenic factors including age, gender, hypertension, diabetes, obesity, smoking, alcohol consumption, dyslipidemia, and hyperuricemia showed no significant difference between the two groups( P>0.05). The level of high-density lipoprotein cholesterol(HDL-C)was lower in the Hp infection group than in the non-Hp infection group( P<0.05). There was no difference in levels of white blood cells, neutrophils, total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), homocysteine, fasting blood glucose, creatinine or uric acid between the two groups( P>0.05). The detection rates of increased carotid IMT and carotid plaques were higher in the Hp infection group than in the non-Hp infection group(65.0% or 89 cases vs.48.7% or 73 cases, 76.6% or 105 cases vs.64.7% or 97 cases, P<0.01 and 0.05). The detection rate of carotid plaques in the Hp-infected group was higher than that in the non-Hp-infected group( P<0.05). The combined detection rate of increased carotid IMT and plaques was higher in the Hp infection group than in the non-Hp infection group(56.2% or 77cases vs.32.7% or 49 cases, P<0.01). Conclusions:Hp infection may play a role in the occurrence and progression of carotid atherosclerosis through initiating abnormal lipid metabolism.Early intervention and treatment may reduce the incidence of carotid atherosclerosis in patients with Hp infection.
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Objective@#To investigate postoperative recurrent pattern of the thoracic esophageal squamous cell carcinoma (TESCC), aiming to provide a basis for the delineation of postoperative radiotherapy volume for TESCC.@*Methods@#Clinical data of 66 TESCC patients who recurred after the radical esophagectomy in Zhongnan Hospital of Wuhan University from 2011 to 2017 were retrospectively analyzed. According to the AJCC 8th edition-defined classification of esophageal carcinoma, regional lymph node stations 1 to 8M were defined as the upper-middle mediastinum region (UMMR), and stations 8Lo, 9 and 15 were defined as the inferior mediastinum region (IMR), stations 16 to 20 were regarded as the upper abdominal lymph node region (UAR).@*Results@#Among all 66 patients, 41 cases (62%) experienced loco-regional recurrence alone, 25 cases (38%) presented with distant metastasis alone. A total of 54 patients with 148 lymph node recurred after treatment. The highest risk region of lymph node recurrence was UMMR (118/148, 80%), after that, followed by UAR (24/148, 17%). With regard to 9 cases of UAR, 6 patients had lower TESCC, and 8 patients (89%) were graded as ≥ pathological stage Ⅲ.@*Conclusions@#The highest risk region of lymph node recurrence is UMMR in TESCC patients undergoing radical esophagectomy, which should be considered as the target volume in postoperative radiotherapy. For patients with lower TESCC ≥ pathological stage Ⅲ, UAR might be the target volume with cautions. Anastomosis and IMR are probably not the routine treatment volumes.
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Primary central nervous system lymphoma (PCNSL) is a type of extranodal non-Hodgkin lymphoma.Whole brain radiation therapy (WBRT) combined with high-dose methotrexate is the standard treatment.Although PCNSL patients are sensitive to radiation therapy,the duration of response is relatively short and it is likely to provoke delayed neurotoxicity,especially in the elderly patients.Reduced-dose WBRT and autologous stem cell transplantation (ASCT) can lower the risk of neutotoxicity,whereas the clinical efficacy remains to be validated.For the elderly patients with PCNSL,application of WBRT in the first-line treatment should be cautiously considered.
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Objective To analyze the clinical efficacy and prognosis of stereotactic body radiation therapy (SBRT) for pulmonary oligometastases.Methods Medical records of 104 patients with SBRT for pulmonary oligometastases in our hospital between 2012 and 2018 were retrospectively reviewed.SBRT was performed by intensity modulated radiation therapy (IMRT) technique before December 2015,and by helical tomotherapy (HT) technique in others.The local control (LC),progression-free survival (PFS) and overall survival (OS) were estimated using the Kaplan-Meier method.Cox-regression was used for univariate analyses and multivariate analyses.The radiotherapy-related adverse events were evaluated by NCICTCAE V4.0.Results The 1-,2-and 3-year LC rates were 86.6％,75.9％ and 72.3％,respectively.The 1-,2-and 3-year PFS rates were 40.9％,28.4％ and 22.1％,respectively.The 1-,2-and 3-year OS rates were 75.9％,53.2％ and 43.53％,respectively.The median OS time was 26.6 months.Multivariate analyses showed that the pathologic type of primary tumor,the volume of lung oligometastases and the carcino-embryonic antigen (CEA) level before SBRT were the independent prognostic factors of LC (x2 =28.66,P＜0.05).The way of tumor progression after SBRT was the independent prognostic factor of OS (x2=40.01,P＜0.05).Meanwhile,there were no significant differences in the LC and OS between HTSBRT and IMRT-SBRT.Radiation pneumonitis was the major adverse event of SBRT (n =25,24.04％).Less than 7％ patients experienced grade 2 and above radiation pneumonitis.Conclusions SBRT shows high local control rates and tolerable adverse events in the treatment of pulmonary oligometastases.There were no significant differences in the clinical efficacy and adverse events between HT-SBRT and IMRT-SBRT,which means they are all suitable for clinical application.
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Objective To investigate postoperative recurrent pattern of the thoracic esophageal squamous cell carcinoma (TESCC),aiming to provide a basis for the delineation of postoperative radiotherapy volume for TESCC.Methods Clinical data of 66 TESCC patients who recurred after the radical esophagectomy in Zhongnan Hospital of Wuhan University from 2011 to 2017 were retrospectively analyzed.According to the AJCC 8th edition-defined classification of esophageal carcinoma,regional lymph node stations Ⅰ to 8M were defined as the upper-middle mediastinum region (UMMR),and stations 8Lo,9 and 15 were defined as the inferior mediastinum region (IMR),stations 16 to 20 were regarded as the upper abdominal lymph node region (UAR).Results Among all 66 patients,41 cases (62％) experienced locoregional recurrence alone,25 cases (38％) presented with distant metastasis alone.A total of 54 patients with 148 lymph node recurred after treatment.The highest risk region of lymph node recurrence was UMMR (118/148,80％),after that,followed by UAR (24/148,17％).With regard to 9 cases of UAR,6 patients had lower TESCC,and 8 patients (89％) were graded as ≥ pathological stage Ⅲ.Conclusions The highest risk region of lymph node recurrence is UMMR in TESCC patients undergoing radical esophagectomy,which should be considered as the target volume in postoperative radiotherapy.For patients with lower TESCC ≥ pathological stage Ⅲ,UAR might be the target volume with cautions.Anastomosis and IMR are probably not the routine treatment volumes.
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Objective The aims of this study were to investigate the correlation between the early stage of severe granulocy-topenia and the curative effect on clinical prognosis of chemotherapy in the first-line EP regimen of small cell lung cancer(SCLC). Methods A retrospective study analysis of 82 cases of first-line EP chemotherapy in patients with SCLC was collected clinical da-ta,according to the time of patients with severe neutropenia.Patients were divided into two groups i.e.early-onset group(the first two cycles appeared Ⅲ-Ⅳ neutrophil decline)and non-early-onset group(Ⅲ~Ⅳneutropenia did not appear or three cycles and lat-er).The main indicators for the observation were objective response rate(ORR),disease control rate(DCR),time to progression (TTP)and overall survival(OS).Kaplan-Meier method and Log-rank test were used to analyze univariate survival and Cox propor-tional hazards model to analyze multivariate survival.Results The effective rates of chemotherapy were 81.8% and 75.5%(P=0.499),and DCRs were 97.0% and 95.9% in early-onset and non-early-onset groups,respectively(P>0.05).The median survival time was 10.4 months and 6.9 months in early-onset and non-early-onset groups,respectively(P=0.001).The median OSs were 22.4 months and 16.0 months in early-onset and non-early-onset groups,respectively(P=0.023).Multivariate surviv-al analysis revealed that smoking index,number of chemotherapy cycles,chest radiotherapy and early-onset severe neutropenia were independent prognostic factors for SCLC(P<0.05).Conclusion Chemotherapy-related loss of early severe neutropenia correlates with the efficacy of chemotherapy in SCLC and is an independent predictor of prognosis in SCLC.
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Objective To evaluate the clinical efficacy and prognostic factors of stereotactic body radiotherapy(SBRT)for pulmonary oligometastases,and to further explore the patients most suitable for SBRT. Methods From 2012 to 2105,51 patients with 76 oligometastatic lung tumors were treated with SBRT.In those patients,27 had primary lung tumors and the others had extrapulmonary tumors. Seven patients had squamous cell carcinoma,thirty-five had adenocarcinoma, and the rest had other types of cancer. The patients received radiotherapy at a dose of 50 Gy in five fractions or 60 Gy in three fractions. Survival analysis was made by the Kaplan-Meier method. A multivariate analysis was made by the Cox model. Results The 1-and 2-year local control rates were 86%(65/76)and 80%(61/76),respectively. The 1-and 2-year overall survival(OS)rates were 80%(41/51)and 55%(28/51),respectively. The median survival time was 30(2-57)months,while the median progression-free survival time was 8(1-32)months. Twenty-one patients had grade 1 radiation pneumonitis(RP),while one patient had grade 2 RP. The multivariate analysis revealed that no more than 2 oligometastatic lung tumors,progression-free interval(PFI), and a performance score(PS)no higher than 1 were independent factors for OS(all P<0.05). Conclusions SBRT is effective and safe for treating pulmonary oligometastases. The number of oligometastatic lung tumors,PFI,and PS are independent prognostic factors for OS. Suitable patients and the appropriate timing of treatment are key to the efficacy of SBRT.
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Objective To study the effect of Interleukin-22 (IL-22) on murine asthmatic airway inflammation and airway remodeling, observe the effect of budesonide on IL-22 of asthmatic mouse model, explore the mechanism of budesonide in the treatment of asthma.Methods Ovalbumin(OVA) was used as an allergen to sensitize and challenge the mice.24 female specific-free (SPF) BALB/c mice aged four weeks were randomly divided into 3 groups:control group, asthma group and budesonide treatment group (BUD group).For Histopathological Examination, HE staining was used to measure the inflammation scores, AB-PAS staining was used to measure the hyperplasia of goblet cells and mucin.Enzyme-linked immunosorbent assay(ELISA) was used to analyze IL-22 levels in bronchoalveolar lavage fluid (BALF).Quantitative Real-time PCR was performed to analyze the effects of budesonide on IL-22 mRNA levels in lung tissue.Results The inflammation scores of asthma group were elevated compared with the control group.An overall change towards less severe asthmatic airway inflammation by the end of the trial was observed in the BUD group.IL-22 levels in BALF were significant decreased after the treatment of budesonide, the mRNA levels of IL-22 were obviously decreased in BUD group, too.A significant positive correlation was observed between the mRNA levels of IL-22 and airway inflammation.Conclusions The increasing IL-22 secretion can lead to the occurrence of airway inflammation of asthma.Budesonide can inhibit the expression of IL-22, thereby Budesonide could inhibit the development of airway inflammation of asthma.
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Objective The objective of this study was to provide a way to assess the nutritional status of patients and to afford targeted nutritional supports during the radiotherapy on the basis of the laboratory parameters related to nutrition and chest muscle size in lung cancer patients at the different time.Methods The laboratory parameters were obtained in a cohort of 160 lung cancer patients who received thoracic radiotherapy in our department from March 2012 to November 2015.Fourteen patients who had complete chest CT scan images during radiotherapy were selected to evaluate chest muscles volume.The Chest muscles and its volume were delineated and calculated by CT scan images.Results The levels of(Hemoglobin)HGB,lymphocyte,total protein and albumin were decreased in different degrees during and after radiotherapy,which had the positive correlation with the number and doses of radiotherapy(P0.05).Conclusion Cancer patients were prone to suffer from malnutrition during radiotherapy.The intake of energy and protein was less than the requirements.We should always take the nutritional status of patients into account and provide targeted nutritional support to improve treatment tolerance and quality of life of patients during radiotherapy.
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Objective To investigate the effect of nerve growth factor ( NGF) combined with exercise train-ing on nerve protection, learning and memory. Methods Twenty-four six-month-old APP/PS1 mice were randomly divided into a control group, an NGF group, an exercise group and an NGF combined with exercise training group ( the combined group) , each of 6. The control group mice were given 50 μL of saline intra-nasally every day, while the NGF group were given 50 μL of NGF solution (0.1 mg/ml) intranasally. The exercise group was given 50μL of saline intra-nasally plus 30 minutes of treadmill exercise daily, while the combined group was given 50μL of the NGF solution and 30 minutes of exercise. The intervention lasted 21 days. Then Morris water maze test was conducted to e-valuate the memory of all groups, and all of the mice were sacrificed and immunohistochemical staining was used to check the levels of APP, Ki-67 expression in cortex and NeuN expression in hippocampus. Results In the place navigation test, significant differences in escape latency were observed among the 4 groups, with the NGF, exercise and combined groups showing average latencies significantly shorter than that of the control group The average latency of the combined group was also significantly shorter than that of the NGF and exercise groups. In the spatial probe, there were significant differences among the 4 groups in the percentage of time spent in the target quadrant. The com-bined group′s average was significantly larger than those of the NGF and exercises groups. The immunochemistry showed that the combined group had better-arranged neurons and more cells compared with the 3 other groups. The average APP protein expression of the combined group was significantly lower than that of the other groups, but their Ki-67-positive and NeuN-positive cells were found to be significantly more numerous. Conclusion NGF combined with exercise can improve learning and memory, at least in APP/ PS1 mice. This might be associated with its reduc-ing the expression of APP and promoting the expression of Ki-67 and NeuN.
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BACKGROUND:Previous research have confirmed that CD34 is closely related to oncogenesis, progress, recurrence, metastasis and drug-resistance of various cancers, but its role in nasopharyngeal carcinoma remains unclear. OBJECTIVE:Tosortcels positive and negative for CD34 in nasopharyngealcarcinoma cel lines and to detect cel proliferation and migration. METHODS:Expressionsof CD34 in nasopharyngeal carcinoma cel lines 5-8F, 6-10B, CNE1 and CNE2 were detected by flow cytometry. And CD34+and CD34-cels were sorted based on cel surfacemarkers for purity identification. Afterwards, proliferation and migrationof CD34+and CD34-celswere detected by MTT assay, colony-formation assay and scratch assay. RESULTS AND CONCLUSION:Al four nasopharyngeal carcinoma cel lines expressed CD34 in 0.1%-0.2%, and the level of CD34 was closely related to the cel growth density. The purity of CD34+cel was more than 98% in the sorted CD34+celpopulations, but no CD34+cels were found inthe sorted CD34-celpopulations.At 1, 3, 5 and 7 daystheproliferation rate of CD34+cel, populationswas significantly higher than that of CD34-cels (P< 0.05). Consistently, thecolony-formation efficiencyof CD34+cel was significantlyhigher than that ofCD34-cels (P< 0.05). Moreover, CD34+cels migrated significantly faster than CD34-cels by scratch assay (P< 0.05). In conclusion, CD34+cels culturedin vitro display higher proliferation and migration capacities, indicating that CD34+celshavethe potential of nasopharyngeal carcinoma stem cels.
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BACKGROUND:Now in mammary stem cellresearch, no proper mammary stem cellmedium is provided to culture mammary stem cells. OBJECTIVE:To create a mammary stem cellmedium and validate its application by isolating Sca-1+mammary stem cells. METHODS:We first used BM medium to culture mammary organoids, and after 6 days, the expression of Sca-1 and vimentin was detected in fibroblasts by immunofluoresence method. Then, we established MaECM medium which arrested fibroblasts growth. After 6 days culture of mammary organoids by MaECM medium, Sca-1+and Sca-1-cellpopulations were sorted out by magnetic sorting and the purity was analyzed by flow cytometry. Sorted 1×104 Sca-1+or Sca-1-cells were transplanted into the bilateral mammary fat pads of four mice, and after 6-8 weeks, the fat pads were harvested for whole-mount immunohistochemical analysis and hematoxylin-eosin staining. RESULTS AND CONCLUSION:After 6 days culture of mammary organoids under BM medium, smal-sized colonies were generated around lots of fibroblasts. Immunofluoresence staining detected strong expression of vimentin and Sca-1 in fibroblasts, indicating that the BM medium is not suitable to isolate Sca-1+mammary stem cell. The MaECM medium promoted the proliferation of mammary epithelial cells whereas arrested fibroblasts growth. After 6 days culture of mammary organoids under MaECM medium and magnetic sorting, the flow cytometry showed that the purity of Sca-1+cellreached 92%and 5%in the Sca-1+and Sca-1-population, respectively. The results from transplantation test showed that six mammary outgrowths were regenerated out of eight injected fat pads in the Sca-1+cells transplantation, but in the Sca-1-transplantation population, one mouse died and the other transplants failed to produce outgrowths. We developed the MaECM medium which promoted the proliferation of mammary epithelial cells whereas arrested Sca-1+fibroblasts growth. Using the medium, we confirmed that Sca-1+mammary cells have capacity of isolating mammary stem cells.
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Objective To investigate the changes in the abilities of invasion and metastasis in surviving human lung adenocarcinoma (A549) cells after X-ray irradiation and the related mechanism.Methods The change in radiosensitivity after X-ray irradiation was determined by colony formation assay.The abilities of invasion and metastasis were evaluated by MTF adhesion assay and Transwell invasion and migration assay in vitro.The mRNA and protein expression of E-cadherin,vimentin,tumor growth factor (TGF)-β1,matrix metalloproteinase (MMP)-2,and MMP-9 was measured by real-time RT-PCR using SYBR Green fluorescence and Western blot.Results The colony formation assay showed that the surviving A549 cells after X-ray irradiation were more resistant to irradiation (ratio of D0 values =0.94).Their abilities of adhesion,invasion,and migration were significantly increased by 1.46 times,1.40 times,and 1.45 times,respectively.In addition,the surviving cells showed enlarged intercellular spaces and had many long pseudopodi.Compared with those in the control group,the mRNA expression levels of vimentin,TGF-β1,MMP-2,and MMP-9 of surviving cells were increased by 1.37 times,2.37 times,1.80 times,and 1.50 times,respectively,the protein expression levels of the above substances were increased by 1.60 times,1.80 times,1.10 times,and 1.20 times,respectively,and the mRNA and protein expression levels of E-cadherin were decreased by 59.4％ and 74.6％.Conclusions The surviving A549 cells after X-ray irradiation have significantly increased abilities of invasion and metastasis.This may be due to radiationinduced TGF-β1 expression increase that in turn promotes epithelial-mesenchymal transition and also due to radiation-induced MMP-2 and MMP-9 expression increase.
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Objective To evaluate the effect of home-made immobilization device with KV-CBCT in lung-SBRT and investigate its clinical use value.Methods Choosing 10 lung tumor patients (half centre type tumor;half peripheral type) random analysis the interfractional and intrafractional setup errors in the SBRT process by this fixed device with KV-CBCT.The concrete method is using Varian's KV-CBCT scans the patients before and after the SBRT each time,then make the registration between the reconstructed 3 d image and the planned CT image (both based on bone landmark),we then obtain the average setup errors in LR,AP and SI directions.Simultaneously,this research make contrastive analysis of setup errors among this fixed device and other fixed devices such as vacuum pad,phantom in body IMRT.All data make one-factor analysis of variance by SSPS 17.0.Results All the setup errors data was gaussian distribution,the centre type interfraction was at (0.01 ±0.32) cm (LR),(-0.08 ±0.38) cm (AP),(0.14 ±0.36) cm (SI) of the cross section,peripheral type interfraction was at (0.01 ± 0.32) cm (LR),(-0.08 ± 0.38) cm (AP),(0.14 ± 0.36) cm (SI) of the cross section (P =0.001).We found out that the average of lung tumor's setup error at all three directions have no significant difference-the largest was the AP directions (P =0.003),the second was the SI direction (P =0.003) and the smallest was the LR direction (P =0.001).The central type has no significant difference at three directions.Compare to the other fixed device,the average setup errors of our device are (0.09 ± 0.33) cm (LR),(-0.10 ± 0.44) cm (SI),(0.17 ±0.35) cm (AP) better than the report at present paper.As the interfraction setup error was small enough by using this fixed device while it has beyond the system algorithm,the registration software of system shows (0.0 ± 0.0 cm).Conclusions The range of lung tumor motion can be cut down obviously and enhance each placement accuracy,repeatability,on SBRT with home-made immobilization device.
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Objective To evaluate a blood test,UniCAP system(screening IgE to food or inhalant allergens,eosinophil cationic protein,ECP)in differentiation the capability of lgE-mediated allergic diseases in children. Methods 26 atopic dermatitis,33 asthma/bronchiolitis,20 allergic purpura, 15 Kawasaki disease patients and 20 normal children were tested total lgE,lgE to food or inhalant allergens,ECP by UniCAP system and compared.Resuits Fx5E positive rate are 73.1%,24.2%,25.0%,13.3%in atopic dermatitis,asthma,allergic purpura,Kawasaki disease respectively,and the Phadiatop positive rate are 23.1%、69.7%、10.0%、6.6%.The serum levels of ECP、TIgE of patients were higher than the mormal controls,with the highest level in asthma, and lowest in Kawasaki disease,the differences were significant(P<0.05).Conclusions Food orinhalant allergens plays an important role in the inflammatory process of asthma,atopic dermatitis,and some role in allergic purpura,Kawasaki disease.Food allergens are more important to atopic dermatitis.and inhalant allergens are more important to asthma.