ObjectiveTo observe the effects of salmeterol fluticasone on the glucose metabolism and bone density of the elderly patients with chronic obstructive pulmonary disease (COPD) combined with type 2 diabetes mellitus (T2DM).MethodsThirty-one patients with COPD combined with T2DM were divided into 2 groups by random digits table,14 cases in control group were given conventional therapy,17 cases in experimental group were given conventional therapy and fluticasone propionate 50/500 μg,twice a day,for 3 months.The fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin,plasma cortisol and bone density respectively before and after treatment were detected.ResultsIn experimental group,the fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin and bone density before treatment was (5.25 ± 0.21 ) mmol/L,(7.14 ± 0.33 ) mmol/L,(5.58 ± 0.26 )％,( 1.96 ± 0.11 ) g/cm2,and after treatment was(5.31 ± 0.27 ) mmol/L,(7.22 ± 0.29 ) mmol/L,(5.67 ± 0.23 )％,(2.03 ± 0.15 ) g/cm2,there was no significant difference (P ＞ 0.05).In control groups,the fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin and bone density before treatment was (5.33 ± 0.35) mmol/L,( 7.26 ± 0.29 ) mmol/L,( 5.62 ± 0.19 )％,( 1.88 ± 0.20 ) g/cm2,and after treatment was ( 5.36 ± 0.31 ) mmol/L,(7.30 ± 0.35 ) mmol/L,( 5.69 ± 0.26 )％,( 1.98 ± 0.17 ) g/cm2,there was no significant difference (P ＞ 0.05 ).There was no significant difference in plasma cortisol before and after treatment in two groups (P ＞0.05).ConclusionInhaling salmeterol fluticasone for elderly patients with COPD combined with T2DM is safe.

Objective To investigate Fascin expression in human lung cancer tissues and its clinical significance.Methods Immunohistochemistry was performed in 62 patients with different histological types and clinical stages of lung cancer and 92 cases with other malignant tumors.Fascin positive rate in each group was calculated and the differences of pathological characters between the two groups were analyzed.Results Fascin expression in lung cancer tissues was significantly higher than that in normal lung tissues(P ＜ 0.05),its expression varied by different clinical stage of lung cancer tissue differentiation.As differentiation degree decreased,Fascin positive rate increased.Fascin expression was independent with age,sex,smoking history(P ＞ 0.05).Fascin expression had no significant difference between lung cancer group and other tumor group(P ＞ 0.05).Conclusions Fascin expression raising might be common in malignant tumors.Fascin expression in lung tissues indicated the possibility of lung cancer.In lung tissue,high expression of Fasein was a sign of poor differentiation and malignant status of lung cancer.

In order to make a report on 90 cases of insomnia treated by the Wei-defensive qi-regulating and brain-strengthening needling technique,as a clinical subject of the multiple center,for assessing the preliminary standardized indications and operating methods and making an initial summary for writing the draft of the technique operation.Methods:By ratio of 1:1,the patients in conformity with the criteria were randomly divided into the experimental group and control group.In the experimental group,45 cases were treated with the Wei-defensive qi-regulating and brain-strengthening needling technique on Baihui (GV 20),Dazhui(GV 14),Shenmai(BL 62),Zhaohai(KI 6),plus ear points.In the control group,45 cases were treated with routine acupuncture on Sishencong(Ex-HN 1),Shenmen (HT 7),Sanyinjiao(SP 6),etc.The therapeutic effects in the two groups were observed and assessed in comparison.Results:The results showed cure in 15 cases.remarkable effect in 27 cases,effect in 42 cases,failure in 6 cases,and the total eriective rate in 94.4% in 90 cases of insomnia.In the three centers,the statistic management was performed based upon PSQI integral and showed significant difference(P＜0.01),and no significant difference among various groups(P＞0.01).Conclusion:The therapeutic effect in the treatment of insomnia by Wei-defensive qi-regulating and brain-strengthening needling technique is better than the control group and the therapeutic effect and methods in various centers are stable.without any deviation.

Objective:To determine the effects of Foxp3-overexpressing lung cancer cells on activated CD4+T lymphocyte.Methods: Stable Foxp3-overexpressing lung cancer cells NCIH-1299,NCIH-hFoxp3,was generated by transfection of NCIH-1299 cells with plasmid pcDNA3-hFoxp3 mediated by Lipofectamine 2000 and by selection with G418,and validated by quantitative PCR and Western blot.The expression levels of IL-8 and IL-10 secreted by NCIH-hFoxp3 and NCIH-control were measured by ELISA.IL-2 secrection by activated human CD4+T lymphocyte which was tested after stimulation with 20% conditioned medium of NCIH-hFoxp3 and NCIH-control cells.The proliferation of activated human CD4+ T lymphocytes was assessed by MTT after coculture with NCIH-hFoxp3 cells.The adhesive ability of activated human CD4+ T lymphocytes was probed with NCIH-hFoxp3 cells by immunocytochemistry.Results: Compared with NCIH-control cells,NCIH-hFoxp3 secreted high level of IL-10 and low level of IL-8.NCIH-hFoxp3 with Foxp3 overexpression significantly suppressed the proliferation,adhesive potential and IL-2 expression by activated CD4+ T cells.Conclusion: Suppression of immune activities of activated CD4+ T cells by Foxp3 overexpression in lung cancer cells may correlate with cytokine IL-8 and IL-10,which can contribute lung cancer progression.

Objective To investigate the effect of gefitinib on mucus hypersecretion by inhibiting epidermal growth factor receptor (EGFR) activity in chronic obstructive pulmonary disease (COPD).Methods Human airway epithelail cell lines 16HBE cells were exposed to cigarette smoke extraction (CSE) to establish the COPD model.EGFR activity was inhibited by tyrosine kinase inhibitor gefitinib.The mRNA expressions of EGFR and MUC5AC were detected by real-time PCR.EGFR,p-EGFR and MUC5AC protein levels were determined by Western blot and ELISA.Results EGFR mRNA level was increased by 12.7％ in CSE and 8.6％ in gefitinib group,but had no significant differences among CSE,gefitinib group and control group (all P＞ 0.05).MUC5AC mRNA levels were enhanced by 141.7％,26.4％ in CSE group and gefitinib group respectively,and there were significant differences among CSE,gefitinib group and control group (all P＜0.05).EGFR protein levels were (600.34±64.58) μg/mg,(632.58±72.94) μg/mg,(584.57±67.39) μg/mg,in control,CSE and gefitinib groups,respectively,and there were no significant differences between groups (all P＞0.05).p-EGFR protein levels were (338.62±45.28) μg/mg,(679.43±78.23) μg/mg,(292.74±59.17) μg/mg in control,CSE and gefitinib groups,respectively.MUC5AC protein levels were(72.80±6.25)μg/mg,(187.00±±10.26)μg/mg,(92.57±8.32)μg/mg in control,CSE and gefitinib groups respectively.Compared with control group,p-EGFR and MUC5AC protein levels were increased significantly in CSE group (both P＜0.05),and had no significant differences in p EGFR and MUC5AC protein levels between control group and gefitinib group.Conclusions CSE may lead to mucus hypersecretion through activating the EGFR-mediated signaling pathways.Gefitinib may inhibit mucus hypersecretion by inhibiting EGFR tyrosine kinanse activity.EGFR may serve as a potential target for COPD.

Objective:To determine the effects of enforced expression of Foxp3 in lung cancer cell with regards to proliferation and tumorgeneity. Methods: A stable subline NCIH-hFoxp3 was established by liopfectamin-mediated pcDNA plasmid transfection carrying exogenous hFoxp3. The growth curve and secrection of IL-8 and IL-10 of NCIH-hFoxp3 were evaluated using MTT and ELISA, respectively. The in vivo tumorigeneity was assessed as well by inoculation of NCIH-hFoxp3 subcutaneously in nude mice. Results:Lung cancer cell NCIH-hFoxp3 with enforced expression of Foxp3 proliferated slowly but exihited increased in vivo tumorgeneity compared with corresponding control subline. In addition,increased expression of hFoxp3 in NCIH-hFoxp3 augmented secretion and at-tenuated secretion of IL-8 and IL-10,respectively. Conclusion:Increased expression of Foxp3 may promote progression of lung cancer cell by change of cellular microenvironment and evasion of immune surveillance.

Objective To investigate the effects of 12-lipoxygenase (12-LO) and angiotensin Ⅱ (Ang Ⅱ) on the CIP/KIP family of cyclin-dependent kinase inhibitors (CKIs) p21,p27 and p57 related to cell hypertrophy.Methods Mesangial cells were treated with high glucose for 24 hours and 48 hours respectively.12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] and Ang Ⅱ were infused to rats by osmotic mini-pump for 1 week and 2 weeks respectively.Rats fed high fat diet were received low dose streptozotocin (STZ) to make type 2 diabetes (DN).The rats were divided into normal control group,DN group,DN+Ang Ⅱ type 1 receptor blocker (ARB) group or 12-LO inhibitor (CDC) group.DN+ARB rats were treated by losartan for 6 weeks,and DN+CDC rats were treated for 8 weeks.Urine albumin and protein expressions of p21,p27 and p57 were detected by ELISA and Western blotting respectively.Glomeruli injury and expressions of p21 and p27 were detected by PAS staining and immunohistochemistry respectively.Results High glucose increased p21 and p27 protein expression in mesangial cells significantly compared with the relative control (all P ＜ 0.05),but had no effect on p57.Ang Ⅱ increased p27 protein expression in gloneruli significantly (P ＜ 0.05),but had no effect on p21 and p57 protein expression.12(S)-HETE increased both p21 and p27 protein expression in glomeruli significantly (all P ＜ 0.05),but had no effect on p57 protein expression.Blood glucose,kidney/body weight,urinary protein,and glomerular p21 and p27 protein expressions were increased in DN group (all P ＜ 0.05) compared with those in control group,with little change of p57 protein expression (P ＜ 0.05).Moreover,glomerular hypertrophy and extra cellular matrix accumulation were observed in DN group.However,urine protein,kidney/body weight,renal injury,but not blood glucose,were decreased in DN+ARB group and DN+CDC group compared with DN group respectively (P＜ 0.05).Further DN+CDC rats had decreased both p21 and p27 protein expressions in glomeruli,but DN+ ARB rats only had decreased p27 protein expression (all P ＜ 0.05).Conclusions 12-LO may induce both p21 and p27 protein expression in DN glomeruli,but Ang Ⅱ may induce only p27 expression.

Objective To investigate the expression of IL-24 in patients with non-small cell lung cancer (NSCLC),and to evaluate its influence on the bioactivity of NSCLC cells. Methods Thirty-nine patient with NSCLC (23 patients with adenocarcinoma and 16 patients with squamous carcinoma) and 17 healthy subjects were enrolled in this study. Serum samples and lung cancer tissues were collected. IL-24 expression in the serum samples was measured using enzyme-linked immunosorbent assay. Its expression at mRNA level in the lung cancer tissues was measured using reverse transcriptional real-time PCR. Adenocar-cinoma cell line A549 and squamous carcinoma cell line NCI-H520 were stimulated with recombinant human IL-24 (10 ng/ml and 100 ng/ml) for 24 hours. Cell proliferation was measured using CCK-8 method. Ap-optosis and cell cycle were measured using flow cytometry. Cell invasion was measured using Transwell as-say. Results Serum IL-24 was significantly elevated in patients with NSCLC in comparison with that in healthy subjects [(144.10±64.43) vs(48.47±18.00) pg/ml]. No significant difference in IL-24 expres-sion was found between patients with adenocarcinoma and squamous carcinoma. IL-24 expression at mRNA level in lung cancer tissues of patients with NSCLC was also significantly increased with an approximately 5-fold enhancement in comparison with that in normal lung tissues. Stimulation with low concentration of re-combinant IL-24(10 ng/ml) promoted the proliferation and suppressed the apoptosis of A549 and NCI-H520 cells. In contrast, high concentration of recombinant IL-24 (100 ng/ml) stimulation notably inhibited the proliferation and enhanced the apoptosis of lung cancer cell lines. No remarkable changes in cell cycle of the two kinds of lung cancer cells in response to IL-24 stimulation were observed. Moreover,low concentration of recombinant IL-24 (10 ng/ml) did not affect the invasion of A549 and NCI-H520 cells,while high concen-tration of recombinant IL-24 (100 ng/ml) significantly inhibited the invasion of lung cancer cells. Conclu-sion IL-24 might influence the bioactivity of NSCLC cells in a concentration-dependent manner. High con-centration of IL-24 might counteract the invasion and metastasis of NSCLC,which is important to prevent dis-ease promotion.

Myeloproliferative neoplasms (MPN) are a group of clonal disorders of hematopoietic stem cells, and JAK2 V617F gene mutation is the main basis for the diagnosis of MPN. Previous studies have shown that BCR-ABL fusion gene and JAK2 V617F gene mutation are mutually exclusive in MPN patients, but in recent years, patients with a double mutation of both genes are often reported. The article synthesizes the relevant domestic and foreign literature in recent years, and reviews the BCR-ABL fusion gene and JAK2 V617F mutation double-positive MPN.

Objectives To evaluate the role of PCSK9 (proprotein convertase subtilisin kexin type 9) on the lipid accumulation and kidney injury of C57BL/6 mice. Methods The 24 h urine of 12 weeks old wide type C57BL/6 mice and PCSK9 knockout (KO) mice were collected through a metabolic cage, followed by perfusion and sacrifice. Urinary microalbumin?to?creatinine ratio (UACr), total cholesterol and triglyceride in kidney tissues were measured by ELISA. BODIPY 493/503 staining and standard transmission electron microscopy (TEM) of kidney tissues was performed for evaluating lipid accumulation and podocyte foot effacement in the kidney. Kidney tissues were also evaluated by PAS stain and TUNNEL stain. PCSK9, podocin and nephrin were quantified through real?time PCR, and the Bcl?2, Bax and cleaved caspase 3 were evaluated by Western blotting. Results Total cholesterol and triglyceride contents were higher in the kidneys of PCSK9 KO mice than controls (P<0.05). The level of lipid accumulation in glomeruli and tubules through BODIPY 493/503 stain, and the amount of lipid drop in TEM were more serious in PCSK9 KO mice. UACr and podocyte foot process effacement were increased, and the transcription of podocin and nephrin were decreased in the kidneys of PCSK9 KO mice (all P<0.05). The expression of Bcl?2 was decreased, and Bax and cleavedcaspase 3 were increased in the kidney samples of PCSK9 KO mice. Conclusion PCSK9 might be reversely involved in lipid homeostasis and accumulation, resulting in injury and apoptosis in the kidneys of C57BL/6 mice.