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Objective To investigate the value of NPM1 and FLT3 gene mutation combined with bone marrow imaging detection in the prognosis judgement of initial treatment cytogenetically normal acute myeloid leukemia (CN-AML). Methods The clinical data of 100 patients (non-M3 type) with primary and initial treatment CN-AML from January 2010 to January 2014 in the Peace Hospital Affiliated of Changzhi Medical College were retrospectively analyzed. All patients were enrolled in the bone marrow imaging examination on the end day of induction treatment or the first day after the end of induction treatment (T time point). Univariate and multivariate prognostic analyses were performed on AML patients according to FLT3 and NPM1 gene status, bone marrow juvenile cell ratio at T time point. Results A total of 100 patients included 36 cases with FLT3 gene mutation and 44 cases with NPM1 gene mutation. The complete remission (CR) rate of CN-AML patients was 13.9% (5/36) and 71.9% (46/64), respectively (P< 0.01), 2-year recurrence-free survival (RFS) rate was 5.6% and 59.8%, respectively (P< 0.01), 2-year overall survival (OS) rate was 15.6% and 66.2%, respectively in FLT3+group and FLT3-group (P<0.01). The median RFS and median OS time in FLT3+ group was 6.9 months and 9.4 months, respectively, and the median RFS and median OS time in FLT3-group had not yet reached. There were no significant differences of all the indexes between the two groups (all P< 0.01). And there were no significant differences in CR rate, 2-year RFS rate and 2-year OS rate between NPM1+group and NPM1-group (all P＞0.05). The CR rate, 2-year RFS rate and 2-year OS rate in NPM1+ FLT3-group were better than those in NPM1- FLT3-, NPM1-FLT3+and NPM1+FLT3+groups; NPM1-FLT3+and NPM1+FLT3+groups had the worst prognosis, and there were no statistical differences in the CR rate, RFS and OS time between the two groups (all P＞ 0.05). The prognosis of the patients in bone marrow juvenile cell ratio < 0.05 group at T time point was better than that in ratio ≥0.05 group (all P< 0.05). CN-AML patients were classified into the good prognosis group (NPM1-FLT3-), the medium prognosis group (NPM1+FLT3-), and the poor prognosis group (NPM1-FLT3+and NPM1+FLT3+) according to the FLT3 and NPM1 genes. The good prognosis group+the ratio of bone marrow juvenile cells at T time point<0.05 group, the good prognosis group+the ratio of bone marrow juvenile cells at T time point≥0.05 group, the medium prognosis group + the ratio of bone marrow juvenile cells at T time point < 0.05 group had no statistical differences in CR rate, 2-year RFS rate and 2-year OS rate (all P ＞0.05). The medium prognosis group + the ratio of bone marrow juvenile cells at T time point ≥0.05 group, the poor prognosis group+the ratio of bone marrow juvenile cells at T time point<0.05 group had the equivalent prognosis, and the average prognosis was moderate; the poor prognosis group + the ratio of bone marrow juvenile cells at T time point ≥ 0.05 group had the worst prognosis. According to Cox multivariate regression analysis, FLT3 gene mutation and the ratio of bone marrow juvenile cells at T time point were independent influencing factors for RFS and OS in CN-AML patients (all P< 0.05). NPM1 was an independent prognosis factor affecting RFS and OS of FLT3-patients (all P < 0.05). Conclusions After induction chemotherapy, the responsiveness and sensitivity of AML patients to chemotherapy regimen can be assessed early and objectively according to molecular genetics and the ratio of bone marrow juvenile cells at T time point, which has a certain value in the prognosis judgement.
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Objective@#To investigate the value of NPM1 and FLT3 gene mutation combined with bone marrow imaging detection in the prognosis judgement of initial treatment cytogenetically normal acute myeloid leukemia (CN-AML).@*Methods@#The clinical data of 100 patients (non-M3 type) with primary and initial treatment CN-AML from January 2010 to January 2014 in the Peace Hospital Affiliated of Changzhi Medical College were retrospectively analyzed. All patients were enrolled in the bone marrow imaging examination on the end day of induction treatment or the first day after the end of induction treatment (T time point). Univariate and multivariate prognostic analyses were performed on AML patients according to FLT3 and NPM1 gene status,bone marrow juvenile cell ratio at T time point.@*Results@#A total of 100 patients included 36 cases with FLT3 gene mutation and 44 cases with NPM1 gene mutation. The complete remission (CR) rate of CN-AML patients was 13.9% (5/36) and 71.9% (46/64), respectively (P < 0.01), 2-year recurrence-free survival (RFS) rate was 5.6% and 59.8%, respectively (P < 0.01), 2-year overall survival (OS) rate was 15.6% and 66.2%, respectively in FLT3+ group and FLT3- group (P < 0.01). The median RFS and median OS time in FLT3+ group was 6.9 months and 9.4 months, respectively, and the median RFS and median OS time in FLT3- group had not yet reached. There were no significant differences of all the indexes between the two groups (all P < 0.01). And there were no significant differences in CR rate, 2-year RFS rate and 2-year OS rate between NPM1+ group and NPM1- group (all P > 0.05). The CR rate, 2-year RFS rate and 2-year OS rate in NPM1+ FLT3- group were better than those in NPM1- FLT3-, NPM1- FLT3+ and NPM1+ FLT3+ groups; NPM1- FLT3+ and NPM1+ FLT3+ groups had the worst prognosis, and there were no statistical differences in the CR rate, RFS and OS time between the two groups (all P > 0.05). The prognosis of the patients in bone marrow juvenile cell ratio < 0.05 group at T time point was better than that in ratio ≥0.05 group (all P < 0.05). CN-AML patients were classified into the good prognosis group (NPM1- FLT3-), the medium prognosis group (NPM1+ FLT3-), and the poor prognosis group (NPM1- FLT3+ and NPM1+ FLT3+) according to the FLT3 and NPM1 genes. The good prognosis group+ the ratio of bone marrow juvenile cells at T time point < 0.05 group, the good prognosis group+ the ratio of bone marrow juvenile cells at T time point ≥0.05 group, the medium prognosis group+ the ratio of bone marrow juvenile cells at T time point < 0.05 group had no statistical differences in CR rate, 2-year RFS rate and 2-year OS rate (all P > 0.05). The medium prognosis group+ the ratio of bone marrow juvenile cells at T time point ≥0.05 group, the poor prognosis group+ the ratio of bone marrow juvenile cells at T time point < 0.05 group had the equivalent prognosis, and the average prognosis was moderate; the poor prognosis group+ the ratio of bone marrow juvenile cells at T time point ≥ 0.05 group had the worst prognosis. According to Cox multivariate regression analysis, FLT3 gene mutation and the ratio of bone marrow juvenile cells at T time point were independent influencing factors for RFS and OS in CN-AML patients (all P < 0.05). NPM1 was an independent prognosis factor affecting RFS and OS of FLT3- patients (all P < 0.05).@*Conclusions@#After induction chemotherapy, the responsiveness and sensitivity of AML patients to chemotherapy regimen can be assessed early and objectively according to molecular genetics and the ratio of bone marrow juvenile cells at T time point, which has a certain value in the prognosis judgement.
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Objective:To investigate the role of transforming growth factor β1 (TGF-β1) in multi-drug resistance in small cell lung cancer and its clinical significance.Methods:The mRNA and protein expressions of TGF-β 1 in H69 and H69AR cells were detected by real-time PCR and Western blot,respectively.After silence of TGF-[β1,the sensitivity of H69AR to drugs was detected by CCK8 assay.The expressions of TGF-β1 in lung cancer and paracarcinoma tissues were examined by QRT-PCR and immunohistochemistry.The relationship of TGF-β 1 expression with clinical pathological features and prognosis of patients was studied.Results:Compared to H69,the mRNA and protein expressions of TGF-β1 in H69AR cells were significantly increased by (5.93±0.47) and (8.49±1.92) folds,respectively (P＜0.01).Transfection ofTGF-β1 siRNA resulted in a decrease of TGF-β1 expression by 70.432％ in H69AR ceils (F=21.20,P＜0.01) and an increase insensitivity to chemotherapeutic agents of H69AR cells (t=4.576,P＜0.05).Compare with the paracarcinoma tissues,the expression of TGF-β1 was significantly increased in small cell lung cancer tissues (t=13.925,P＜0.01),which was closely related with clinical stage,chemosensitivity and overall survival (all P＜0.05),but not related with gender,age (both P＞0.05).Conclusion:TGF-β1 is involved in the regulation of small cell lung cancer multidrug resistance,which may be a potential marker to evaluate the chemosensitivity and dinical prognostic for small cell lung cancer.
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Objective To compare the effect of enteral nutrition liquid with different dietary fiber content on serum protein in patients with stress hyperglycemia.Methods A total of 90 patients with stress hyperglycemia were randomly divided into three groups:enteral nutritional suspension (SP) group (Peptisorb),enteral nutritional suspension (TPF-FOS) group (Jevety) and enteral nutritional suspension (TPF-D) group (Glucerna),each group consisting of 30 patients.In the three groups,prealbumi (PA),serum albumin (ALB),retinol binding protein (RBP),transfer-rin (TRF) and hemoglobin (Hb) were continuously measured on the 1st,2nd,3rd,4th,5th,6th and 7th day.Data were analyzed by SPSS 15.0.Variances on time and group were analyzed by repeated measures of general linear model.Results The PA,ALB,RBP,TRF and Hb were significantly different among Peptisorb group,Jevety group and Glucerna group (P＜ 0.05).The PA,ALB and Hb which were determined in different time,were not significantly impacted by Peptisorb,Jevety or Glucerna,and not significantly changed with time.The RBP and TRF which were determined at different time,were not impacted by Peptisorb,Jevety or Glucema,but time× group from RBP and TRF which were determined in different time,were significantly impacted in Peptisorb group,Jevety group and Glucerna group (P＜ 0.01).Conclusion The content from different dietary fiber significantly affects serum protein in patients with stress hyperglycemia,and TPF-D has the most significant effect on serum protein.
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Objective:To explore the expression of miR-139-5p in small cell lung cancer (SCLC)tissue and its clinical significance, and to clarify the role of miR-139-5p in the occurrence and development of SCLC. Methods:The biological function of miR-139-5p was examined by cell growth,apoptosis and cell cycle analysis. The expressions of miR-139-5p in 50 cases of cancer tissue and paracarcinoma normal tissue were examined by QRT-PCR.Combined with the clinical data,the role of miR-139-5p in clinic was anzlyzed.Results:The expression level of miR-139-5p in SCLC tumor tissue was lower than that in normal lung tissue (P 0.05).The expression level of miR-139-5p in the patient at LD stage was lower than that of the patients at ED stage (P <0.01).The expression level of miR-139-5p in the resistant patients was higher than that of the patients sensitive to chemotherappy (P <0.01).The expression level of miR-139-5p in the survival patients was lower than that in the death patients (P <0.01).Cox regression analysis indicated that miR-139-5p expression and disease stage were the independent prognostic factors for SCLC.Conclusion:miR-139-5p in participates in the occurrence and development of SCLC by inhibiting the cell proliferation,promoting apoptosis and inducing the cell cycle arrest;it may be used as a target gene to evaluate the prognosis of SCLC patients.
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<p><b>OBJECTIVE</b>To detect the activity of autophagy and explore the impact on survival and proliferation of HEL cells and hematopoietic cells of polycythemia vera （PV） patients with JAK2 V617F mutation.</p><p><b>METHODS</b>Flow cytometry, AO staining and Western blot methods were used to detect the autophagy activity and the expression of LC3-Ⅱ protein of JAK2 V617F+ HEL cells and hematopoietic cells of 12 newly diagnosed PV patients with JAK2 V617F mutation. HEL cells and bone marrow cells of 3 PV patients were treated with rapamycin or 3-MA to induce and inhibit autophagy, respectively. CellTiter Glo(R) method was used to detect the proliferation activity of cells.</p><p><b>RESULTS</b>There was higher level of mean LC3-Ⅱ fluorescence intensity in HEL cells (159 389 ± 29 001) than that in K562 cells (96 047 ± 24 134) (P=0.044). The formation of autophagosome in HEL cells is more than that in K562 cells detected by microscope. What's more, the level of mean LC3-Ⅱ fluorescence intensity in 12 PV patients' myeloid cells (92 842 ± 4 250) was higher than that of 15 healthy volunteers (86 633 ± 2 504) (P=0.001). The expression of LC3-Ⅱ protein was higher in PV patients' peripheral blood cells than that in healthy volunteers detected by Western blot. After treated with rapamycin 12, 24, 48 h, the activity of autophagy in HEL cells and bone marrow cells of 3 PV patients were increased and the proliferation activity was higher than the control group, the proliferation activity at 48 h were (101 413 ± 3 720), (18 744 ± 1 015), respectively. However, after treated with 3-MA 12, 24, 48 h, the activity of autophagy was decreased and the proliferation activity was lower than the control group, the proliferation activity at 48 h were (5 732 ± 166), (5 371 ± 56), respectively.</p><p><b>CONCLUSION</b>There is high basical activity of autophagy in JAK2 V617F+ HEL cells and hematopoietic cells of PV patients with JAK2 V617F mutation. Up-regulated autophagy promotes proliferation of JAK2 V617F⁺ HEL cells and bone marrow cells of PV patients with JAK2 V617F mutation. Decreased autophagy inhibits proliferation of JAK2 V617F+ HEL cells and bone marrow cells of PV patients with JAK2 V617F mutation.</p>
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Humans , Autophagy , Cell Proliferation , Janus Kinase 2 , Mutation , Polycythemia VeraABSTRACT
Objective To investigate the proliferation inhibition and the apoptosis induction effect of brucine on human chronic myeloid leukemia cell line HL-60 cells.Methods HL-60 cells were exposed to various dosages of brucine 24,48,72 h respectively,MTT method was used to assay the growth inhibition effect of brucine on HL-60 cells and the IC50 of brucine was evaluated at the same time.The morphology was observed by AO/EB stains.The cell apoptosis and cell cycle was tested by flow cytometry with Annexin V-FITC/PI double staining and PI labeling respectively.Results The results indicated that the brucine displayed significant anti-proliferative effect on HL-60 cells in a dose-and time-dependent manner,with IC50 value of 211.8 μg/ml(24 h),107 μg/ml(48 h),83 μg/ml(72 h)respectively.The most significant inhibition was observed at 320 μg/ml for 48 h.In this condition,apoptosis morphology was induced by brucine with nuclear chromatin condensation,most of the nuclears were orange stained and condensation-like or bead-like,which was consistent with the Annexin V-FITC/PI results.The cell apoptosis rates were(2.1±1.1)％,(21.3±1.2)％,(38.6±1.3)％,(58.5±4.1)％,(75.3±0.87)％and(66.2±0.75)％in different dose of brucine,respectively.At the same time,the cell cycle analysis results showed that the cell ratio in G0/G1 phase was decreased while in G2/M and sub-G0 phases was increased,comparing with blank control group.Conclusion Brucine can inhibit cell growth dramatically,which may be related to the cell apoptosis and the cell cycle arrest.
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Objective To investigate the distribution and antibiotic resistance of pathogens isolated from inpatients with urinary tract infections.Methods A retrospective analysis was carried out on 1033 strains of pathogens isolated from urine culture in patients with urinary tract infections in Zhejiang Xiaoshan Hospital during January 2009 and December 2011.Urine specimens were cultured with Uricult,and K-B method was used for drug susceptibility test,WHONET 5.6 software was used to analyse drug susceptibility test.Results Among 1033 strains of pathogens,681 (65.9％) were gram-negative bacteria,197 (19.1％) were gram-positive bacteria,and 155 (15.0％) were fungi.The three most prevalent bacteria were Escherichia coli (402 strains,38.9％),Klebsiella pneumoniae (74 strains,7.2％) and Candida albicans (64 strains,6.2％).60.7％ (244/402) of Escherichia coli and 45.9％ (34/74) of Klebsiella pneumoniae were extended-spectrum β-lactamases (ESBLs) positive.Escherichia coli and Klebsiella pneumoniae were susceptible to imipenem,meropenem,cefoperazone/sulbactam,piperacillin/tazobactam and amikacin.Enterococcus and staphylococcus were susceptible to vancomycin,linezolid and furadantin.Candida was sensitive to flucytosine,voriconazole and amphotericin B.Conclusion Gram-negative bacteria mainly E.coli is the predominant pathogen to urinary tract infections in this group of patients.Regular analysis and monitoring of pathogen species and drug resistance is important for rational use of antibiotics.
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Objective To analyze the features of clinical diagnosis and treatment of duodenal gastrointestinal stromal tumors(GIST).Methods Retrospective analysis was performed on clinical data of 5 cases of duodenal GIST treated in our hospital between April 2002 and May 2009.Results The duodenal GIST was mainly located in the 2nd(3/5)and 3rd portion of duodenum(2/5).Clinical diagnosis of the disease mainly depended on barium meal examination, gastrointestinal endoscopy, and CT scanning.Two patients were treated with pancreaticoduodenectomy, 1 with segmental duodenectomy and 2 with local resection.After operation, 2 patients had recurrence and 1 of them underwent adjuvant therapy with Gleevec.Conclusion Surgical resection is the only effective therapeutic method for duodenal GIST.Various surgical procedures are mainly determined by the location and size of the tumors.For patients with a high degree of pathologic grade, adjuvant therapy with Gleevec is necessary.
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OBJECTIVE To explore the relationship between cytokines(IL-1?,mIL-2R,IL-10) and hepatatis B virus(HBV) serum markers in patients with chronic hepatitis B(CHB).METHODS The serum level of IL-1?,mIL-2R and IL-10 in 65 patients with CHB was measured with chemiluminescence immunoassay(CLIA).RESULTS Of the patients with positive and negative HBeAg,the serum IL-1? and IL-10 levels were significantly higher than normal controls(P