[MiR-217 Targeting PI3K/Akt Pathway Enhances Sensitivity of Adriamycin to Acute Myeloid Leukemia].

OBJECTIVE: To investigate the effects of miR-217 on proliferation and adriamycin sensitivity of acute myeloid leukemia (AML) cells. METHODS: The mimic NC and miR-217 mimic vectors were constructed and transfected into HL-60 cells, and transfection efficiency was detected by qPCR. The cells were treated with different concentrations of adriamycin for 24 h and 48 h. CCK-8 assay was used to detect the chemical sensitivity of adriamycin and screen the optimal concentration and time of adriamycin treatment. Cells were divided into control group, mimic NC group, miR-217 mimic group, adriamycin group and miR-217 mimic+adriamycin group. Apoptosis was detected by flow cytometry, and the expressions of miR-217, PI3K and Akt3 were detected by qPCR. Western blot was used to detect the expression of PI3K/Akt pathway proteins PI3K, Akt3 and apoptosis proteins Bcl-2, Bax, and double luciferase was used to verify the relationship between miR-217 and Akt3. RESULTS: MiR-217 mimic could enhance the sensitivity of HL-60 cells to adriamycin. The optimal concentration and treatment time of adriamycin were 160 ng/ml and 48 h, respectively. Compared with control group, apoptosis rate, miR-217 and Bax protein levels were significantly increased in miR-217 mimic and adriamycin groups (P < 0.01), while Bcl-2 protein, PI3K, Akt3 mRNA and protein levels were significantly decreased (P < 0.01). Compared with adriamycin group, apoptosis rate, miR-217 and Bax protein levels were significantly increased in miR-217 mimic+adriamycin group (P < 0.01), while Bcl-2 protein, PI3K, Akt3 mRNA and protein levels were significantly decreased (P < 0.01). Dual luciferase assay showed that there was a targeted regulatory relationship between miR-217 and Akt3. CONCLUSION: MiR-217 regulates the PI3K/Akt pathway targeting Akt3, inhibits cell proliferation, promotes cell apoptosis and enhances the sensitivity of adriamycin to AML cells.

[Effect of Long Non-Coding RNA LINC01268 on the Malignant Biological Behaviors of Acute Myeloid Leukemia Cells].

OBJECTIVE: To investigate the effect of long non-coding RNA LINC01268 on apoptosis of acute myeloid leukemia (AML) cells and related mechanisms. METHODS: The expression levels of LINC01268 and miR-217 in peripheral blood samples from AML patients and AML cell lines HL-60 and KG-1 were detected by qRT-PCR. HL-60 cells were divided into pcDNA3.1-NC, pcDNA3.1-LINC01268, si-NC, si-LINC01268, miR-NC, miR-217 mimics, si-LINC01268 + inhibitor-NC and si-LINC01268+ miR-217 inhibitor groups. The mRNA expressions of LINC01268 and miR-217 were detected by qRT-PCR. The targeting relationship between LINC01268 and miR-217 was detected by dual-luciferase reporter assay. Cell viability was detected by CCK-8 assay. Cell cycle distribution and apoptosis were detected by flow cytometry. The expression of cell cycle and apoptosis-related proteins p21, Bcl-2, Bax, caspase-3 and PI3K/AKT signaling pathway-related proteins were detected by Western blot. RESULTS: The expression of LINC01268 in peripheral blood samples of AML patients and AML cell lines HL-60 and KG-1 was increased (P < 0.05), and the expression of miR-217 was decreased (P < 0.05). Compared with si-NC group and miR-NC group, the viability of HL-60 cells was decreased in si-LINC01268 group and miR-217 mimics group (P < 0.05), the proportion of cells in G1 phase and apoptosis rate were increased (P < 0.05), the protein expression levels of p21, Bax and caspase-3 were increased (P < 0.05), while the protein expression level of Bcl-2 was decreased (P < 0.05). LINC01268 targeted and negatively regulated the expression of miR-217, and inhibiting the expression of miR-217 partially reversed the effects of LINC01268 interference on the viability, cell cycle and apoptosis of HL-60 cells. Interference with LINC01268 could inhibit the activity of PI3K/AKT signaling pathway. Inhibiting the expression of miR-217 could partially reverse the inhibition of LINC01268 interference on PI3K/AKT signaling pathway. CONCLUSION: LINC01268 is highly expressed and miR-217 is lowly expressed in AML cells. LINC01268 can promote the activity of PI3K/AKT signaling pathway, increase the survival rate and inhibit the apoptosis of AML cells by targeting miR-217 expression.

International approaches to coordinated care delivery systems / 中华医院管理杂志

This paper summarized practices and experiences of other countries in coordinated care delivery system building,and described the care delivery systems used in various healthcare fundraising patterns.It is found that most countries have defined duties and service coverage of healthcare institutions at all the levels and measures to ensure rational patient flow.In the end,the paper concluded found experiences of these countries and inspirations for China.

Clinlcal effect observation on the treatment of stomach cancer with self-designed Chinese formula and chemotherapy / 国际中医中药杂志

Objeetive To investigate the clinical efficacy of treating stomach cance belonging to spleen and stomach deficient cold type with self-designed formula and chemotherapy.Methods 100 patients of stomach cancer belonging to stomach and spleen deficient cold were randomly grouped into a control group and a treatment group,with 50 cases in each group.The control group was only treated with chemotherapy,and the treatment group was additionally treated with self-designed Chinese formula on the basis of the control group.After 3 months' treatment,the curative effect,the improvement of life quality,the changes of body weight,the improvement of peripheral blood and immune function,and the occurrence of adverse reaction were observed.The survival rate of patients after 1,3 and 5 years' treatment was investigated.Results ① Short term curative effect:the total effectiveness of the control group and the treatment group was 60.0％ (30/50) and 30.0％ (15/50) respectively,with the treatment group being better than the control group(Z=-2.100,P＜0.05);② life quality:theimprovement rate of life quality in the treatment group and the control group was 64.0％ (32/50) and 20.0％ (10/50) respectively,with the treatment group being better than the control group (Z=-3.259,P＜0.05); ③ body weight:the improvement rate of body weight in the treatment group and the control group was 24.0％ and 16.0％ respectively,and there was no significant difference (Z=-1.815,P＞0.05) ; ④ peripheral blood:the peripheral blood abnormality rates of treatment group was obviously lower than the control group (Z=-4.286,P＜0.05) ;⑤ immune function changes:CD4+、CD8+、and the activity of NK cell of two groups after the treatment [the treatment group was (40.8 ± 4.1)％,(26.1-3.2)％,(83.57-2.52)％ respectively; the control group was (31.7±2.9)％、(33.8±3.0)％、(92.88±3.83)％ respectively] wasobviously improved compared with the same group before the treatment [the treatment group was (34.2±3.5)％,(31.0±2.0)％,(85.01±2.92)％ respectively; the control group was (34.0±3.2)％,(30.9±2.2)％,(85.02± 3.48) ％ respectively].The immune indexes (CD4+、CD8++、CD4+/CDg+and the activity of NK cell)in the treatment group presented significant difference (P＜0.01) than the control group after the treatment.⑥adverse reaction:compared with the control group,the occurrence rate of adverse reaction of the treatment group was obviously lower than the control group,and there was significant difference (Z=-5.297,P＜0.05).⑦survival rate:the survival rate of patients after 1,3 and 5 years' treatment in the treatment group was better than the control group,and there was significant difference (x2=5.263,12.96,7.895,P＜0.05).Conclusion The combination of self-designed Chinese formula and chemotherapy could effectively relieve the clinical symptoms,improve the immunity,reduce the occurrence of adverse reaction,and improve the life quality of patients with stomach cancer belonging to stomach and spleen deficient cold type.

Prognostic significance of a newly proposed grading and scoring system in stage I pulmonary adenocarcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To evaluate the prognostic significance of a new grading and scoring system (based on the new IASLC/ATS/ERS classification) in stage I pulmonary adenocarcinoma, as compared with the WHO grading system.</p><p><b>METHODS</b>The clinicopathologic characteristics of 125 patients with stage I pulmonary adenocarcinoma primarily treated by surgical resection were reviewed retrospectively. All cases were classified according to the new IASLC/ATS/ERS classification and graded into three prognostic groups based on the new classification, the Sica scoring system and the WHO grading system, respectively. The differences in prognosis of the three groups were analyzed.</p><p><b>RESULTS</b>There was a statistically significant correlation between the new grading system and the WHO grading system (P = 0.000). Both of them showed negative correlation with overall survival. The new scoring system however better correlated with disease recurrence and/or metastasis (P = 0.855, P = 0.073 versus P = 0.011). According to univariate Log-rank test, the prognosis correlated with tumor size (P = 0.004), clinical stage (P = 0.000), the WHO grading (P = 0.020), the new grading system (P = 0.000), the new scoring system (P = 0.000), vascular invasion (P = 0.021), and recurrence and/or metastasis (P = 0.000). The Cox regression analysis demonstrated that clinical stage (P = 0.014), the new grading system (P = 0.047), the new scoring system (P = 0.043), and recurrence and/or metastasis (P = 0.018) were significantly independent poor prognostic factors.</p><p><b>CONCLUSIONS</b>The new grading and scoring system shows good correlation with the WHO grading system. Compared with the WHO grading system, the new scoring system based on the new IASLC/ATS/ERS classification provides valuable information in categorizing stage I pulmonary adenocarcinoma cases with different risks of disease recurrence, tumor metastasis and prognosis.</p>

Establishment of a radioresistant human lung cancer cell subline and its mechanism of radioresistance / 中华放射医学与防护杂志

Objective To establish a radioresistant cell subline from a human A549 lung cancer cell line and investigate the mechanism of radioresistance. Methods Two proposals were applied for the non-small cell lung cancer A549 cells irradiated with X-rays:A group of A549 cell line was irradiated five times, the fractionated dose was 600 cGy, and the other group was exposed 15 times, the fractionated dose was 200 cGy. After the completion of irradiation, two monoclones were obtained from the survival of cells and named the subline A549-S1 and A549-S2. The radiosensitivity and cell cycle distribution of these two clones,together with its parental A549 cells were measured by clone formation assay and flow cytometry.The mRNA and protein levels of Notch1 in A549 cell line and the sublines were determined by RT-PCR and Western-blots. Results Compared with the parental A549 cells, A549-S1 cells showed significant resistance to radiation with D0, Dq and N values increased, and a broader initial shoulder as well as 1.38-fold increased value of SF2. The A549-S1 subline also showed higher percentage of cells in S phase and G2/M phase, but lower percentages in G0/G1 phase (P<0.05). The expression of Notch1 in A549-S1 was enhanced obviously than in A549 cells. But for A549-S2 the radioseasitivity was slightly increased compared with the parental cells with D0, Dq and N values decreased and a curve initial shoulder. The ratio of cells in S and G0/G1 phase ratio was lower than that in parental A549 cells, but that in G2/M phase ratio was higher significantly (P<0.05) .The expression of Notch1 had no marked change compared to A549 cell. Conclusions The radioresistance of the A549 cell subline is correlated with the irradiation program. The cell subline shows a different cell cycle distribution from their parental line. The cell cycle distribution has a close correlation with the expression of Notch1.

[Prognostic analysis of chronic myeloid leukemia with Cox model and the Sokal, Hasford score].

This study was aimed to analyze the prognostic factors of the patients with chronic myeloid leukemia (CML). Survival curve and survival rate of 204 patients with CML were estimated with Kaplan-Meier method and Logrank respectively. Univariate and multivariate analysis of prognostic factors carried out by Cox's regression model. The Sokal and Hasford score were used to discriminate the relative risk of Hu and IFN group. The results showed that among the 204 patients, the median survival time was 50 (32-65) months, and 5 year survival rate was 32.3% (95% CI, 23.7%-42.6%). The median survival times of IFN and Hu group were 56 (41-67) and 41 (19-56) months, and 5 year survival rates were 45.4% (95% CI, 37.5%-54.2%) and 26.8% (95% CI, 21.6%-33.3%) (P < 0.001) respectively. From the Cox stepwise regression model, Ph chromosome negative, high LDH, low Hct, percentage of peripheral blood basophils > or = 10%, marrow blasts + promyelocytes > or = 10% and presence of nucleated RBCs was associated with poor prognosis, and the treatment also played an important role in CML. According to the Sokal score, the high, intermediate and low risk rates of Hu group were 72.9%, 21.5% and 5.6%, the median survival time reached 34 (23-49) months, 43 (32-58) and 50 (38-62) months respectively; while censored by the Hasford score, the high, intermediate and low risk rates of IFN group were 17.6%, 25.1% and 57.3% respectively, the median survival time was 44 (33-57), 56 (45-70) and 66 (52-76) months respectively. It is concluded that Ph chromosome, concentration of LDH, percentage of Hct, peripheral blood basophils, marrow blasts, promyelocytes, presence of nucleated RBCs and treatment are the most important prognostic factors for CML. The Sokal score can not discriminate the relative risk of Hu group well, while the Hasford score can discriminate the relative risk of IFN group.

Role of tyrosine protein kinase in proliferation and hypertrophy in vascular smooth muscle cells of spontaneously hypertension rats / 中国病理生理杂志

AIM: To explore the relationship between proliferation, hypertrophy of vascular smooth muscle cells(VSMC) and platelet-derived growth factor-AA(PDGF-AA) and PDGFR-? expression in spontaneously hypertension rats(SHRs) and the role of tyrosine protein kinase (PTK) in it. METHODS: 1. The difference of PDGF-AA, PDGFR-? and PDGFR-? expressions in SHR/Wistar-kyoto rat(WKY)-VSMC was examined by Western blot. 2.The effect of PTK inhibitor (genistein) on proliferation and hypertrophy of SHR-VSMC induced by PDGF-AA was observed by Western blot and incorporation. RESULTS: 1.PDGF-AA and PDGFR-? expression markedly increased in SHR-VSMC compared with WKY-VSMC. There was no difference in PDGFR-? expression levels between SHR and WKY-VSMC. 2.Dose-dependent PDGF-AA-stimulated PCNA expression and incorporation were observed in SHR-VSMC. 3. Genistein inhibited PCNA expression and incorporation induced by PDGF-AA in a dose-dependent manner in SHR-VSMC. CONCLUSIONS: Spontaneous increasing expression of PDGF-AA and PDGFR-? in spontaneously hypertension rats(SHRs) may be one of the important factors in vascular reactivity and vascular modeling mediated through proliferation and hypertrophy in SHR-VSMC. Tyrosine protein kinase plays an important role in this process.