Objective To examine the role of SPHK-1/S1P and NFκB p65 signal pathway in non-small cell lung cancer (NSCLC)drug-resistant cells.Methods The drug-resistant cell line of lung cancer H460/DDP was constructed and its biological characteristics were identified successfully.The expression of SPHK-l was tested by RT-PCR and Western blot methods.S1 P and some proteins related to NFκB pathway were studied by Western blot. Results The drug-resistant lung cancer cell line H460/DDP was constructed and its drug-resistant ability was evaluated (IC50H460/DDP = 50.62μg/mL, RIH460/DDP = 2.95 ). Cisplatin at a concentration of 10 - 80 μg/mL significantly decreased cell death of drug-resistant cell line (P<0 .01 ).Western blot assay analysis showed that overexpressions of SPHK-l,S1 P and NFκB p65 were significantly higher in drug-resistant cell line than in their parent cell line H460 (P=0.0415,P=0.0465,P=0.0218).RT-PCR method revealed that SPHK-1 mRNA was overexpressed in drug-resistant cell line compared with that in their parent cell line H460 (P<0.05).More NFκB p65 protein in cell nucleus was expressed in drug-resistant cells than in parent cells.Conclusion SPHK-1/S1P and NFκB p65 signal pathway may play an improtant role in the drug-resistant H460 cell line in non-small cell lung cancer.

Objective To investigate the mRNA and protein expression of E3 ubiquitin ligase Iduna in non-small-cell lung cancer tissue and para-neoplastic lung tissue,and the correlation of the Iduna expression with clinicopathological factors and prognosis.Methods The expression levels of the Iduna mRNA and protein in non-small-cell lung cancer tissue and para-neoplastic lung tissue were determined by reverse transcriptase-polymerase chain reaction (RT-PCR),Western-Blot and immunohistochemistry respectively,and the correlation of the Iduna expression with clinicopathological factors and prognosis was analyzed.Results RT-PCR and Western-Blot showed the expression levels of the Iduna mRNA and protein in non-small-cell lung cancer tissue (0.468 ± 0.086 and 2.554 ± 0.544) were significantly higher than those in para-neoplastic lung tissue (0.203 ± 0.070 and 1.570 ± 0.316),there were statistical differences (P ＜ 0.05).Immunohistochemistry results showed that Iduna was negative expression in the alveolar epithelium cells,negative or weak positive expression in normal bronchial and positive expression in different degrees in the non-small-cell lung cancer tissue.Iduna high expression rate was negative correlation with tumor differentiation (P =0.002),Iduna high expression rate was positive correlation with large tumor size (P =0.044),TNM staging (P=0.015) and lymph node metastasis (P=0.009).Iduna high expression of I stage non-small-cell lung cancer patients was correlated with poor post-operative survival (P =0.016).Conclusions High expression of Iduna may be related to the process of invasion and metastasis of nonsmall-cell lung cancer.It is possible that Iduna serve as potential markers for predicting prognosis in nonsmall-cell lung cancer.

Objective To conduct a pilot study on genome-wide in vivo protein-RNA interactions in E.coli.Methods Bacterial lysate was treated with RNase before the RNA fragments protected by proteins were extracted from treated lysate and used to construct cDNA library that was applied to high-throughput sequencing .Finally, the transcripts bound by proteins were obtained by bioinformatics analysis .Results A total of 3193 transcripts were obtained , including 2234 mRNAs, 47 sRNAs, 39 tRNAs, 11 rRNAs, and 862 intergenic regions .Conclusion Some information of transcripts interacting with proteins in E.coli is acquired , which will facilitate further studies of protein-RNA interactions .

Objective To evaluate the response of the lung tumor xenografts in nude mice to antiangiogenic treatment from perspectives of anatomic,vessel function,cellular and molecular level using the multimodality imaging techniques including optical imaging,dynamic contrast enhanced MRI(DCE-MRI)and diffusion weighted imaging(DWI).Methods The green fluorescent protein(GFP)was transplanted labeled using GFP-expressing NCI-H460 cells.After the transfection of GFP,NCI-H460 cells were implanted subcutaneously into nude mice.Ten days after implantion,12 nude mice whose tumor xenografts grew to 0.5-1.0 cm in the maximum diameter were randomly divided into 2 groups,and injected with phosphate-buffered saline and recombinant human endostatin respectively.Then the nude mice in the two groups underwent optical imaging,DCE-MRI and DWI.The volumes,photon counts,the quantitative MR vessel functional parameters including volume transfer constant(Ktrans),rate constant(Kep),volume of extravascular extracellular space(Ve)and maximum area under the enhancement curve(iAUC),and apparent diffusion coefficient(ADC)values of the tumors were recorded.Then tumors were collected and observed using the transmission electron microscopy and pathology examination,including HE staining,microvessel density(MVD)and the expressions of vascular endothelial cell growth factor(VEGF).The Kep and VEGF expressions in experimental group and control group were compared with x2 text,and other values were compared with t test.The Pearson and Spearman test were used for analyzing the correlation of values in the two groups.Results Seven days after inoculation,the fluorescence signals were detected and grew with the growth of the tumors.On the 7 day after starting therapy,the photon counts of experimental group and control group were(2.51 ± 2.43)× 1010(photon/sec)and(5.77 ± 3.25)× 1010(photon/sec),respectively with no significant differences(t =1.964,P ＞0.05).Two sample t test showed that the tumor volumes in experimental group were smaller than those in control group[(365 ± 56)vs(987 ± 265)mm3,t =0.001,P ＜ 0.01].There was a positive correlation(r =0.673,P ＜ 0.05)between the photon counts and the volumes of the tumors.The mean Ktrans,Kep,Ve and iAUC of experimental group were:(0.055 ±0.012)min-1,0.335(0.184—0.894)min-1,0.297 ± 0.041 and 7.334 ± 3.930,and those for control group were:(0.117 ± 0.027)rin-1,0.417(0.324-1.736)min-1,0.326:±:0.062 and 13.280 ± 4.245.There were significant differences of Krans and iAUC(t =5.155,2.518,P ＜ 0.05)between experimental group and control group.And there was a positive correlation(r =0.715,P ＜ 0.0 1)between the values of iAUC and MVD,but not the expressions of VEGF(r =0.484,P ＞ 0.05).The values of ADC in experimental group were higher than that in control group[(791 ± 38)× 10-6 vs(737 ± 43)×10-6 mm2/s],and there were significant differences(t =-2.299,P ＜ 0.05).Two sample t test showed that the MVD in experimental group were lower than that in control group[(11.9 ± 4.8)vs(19.2 ±4.3)item/hpf,t =2.774,P ＜ 0.05].The VEGF expressions in experimental group were lower than that in control group(x2 =4.000,P ＞ 0.05).It was observed that some cells in experimental group had degenerated and apoptotic signs by the electron microscopy.Conclusions Evaluating the response of lung tumor xenografts to antiangiogenic treatment at anatomical,vessel functional,cellular and molecular level using the multimedality imagings is applicable.And it will be in favour of evaluating the therapeutic effect promptly.

Objective To explore the effect of carvedilol on ACE2 gene and protein expression in chronic heart failure rats after myocardial infarction.Methods The heart failure model was induced by acute myocardial infarc- tion (AMI) through ligating the left anterior descending coronary artery.One month after operation,rats were randomized to receive placebo or carvedilol 2 mg/(kg?d),by gavage.Sham-operated rats were used as the control group.Hemodynamies,body mass and left ventrieular mass index,plasma and myocardial level of angiotensin Ⅱ were determined.ACE2 gene and protein expression was assessed by using RT-PCR and Western Blot.Results The mortality of placebo and Carvedilol groups were 20%,compared with 0% in sham operated rats.Carvedilol significantly improved LVEDP,LVSP,+dp/dt_(max) and-dp/dt_(min) in CHF rats but all the hemodynamics data were still inferior than that of controls.Plasma and myocardial angiotensin Ⅱ level were increased significantly in CHF placebo rats than those of control rats (plasma Ang Ⅱ:CHF:194?19 vs controls:132?15 ng/L,myocardium Ang Ⅱ:CHF:6.7?0.4 vs control:3.8?0.3 ng/g,P

BACKGROUND:There are various methodstoinduceadipogenic differentiation ofbone marrow mesenchymal stem cels, and the main componentfor adipogenic induction isindomethacin or rosiglitazone. However, there is a lack of comparative studyonthe induction efficiency and mechanism among these methods.
OBJECTIVE:Tocompare the adipogenic responses ofhuman bone marrow mesenchymal stem celsto different induction methods, and to analyze the mechanismunderlyingdifferent induction efficiency.
METHODS:After isolation and purification,the adipogenic abilitiesof human bone marrow mesenchymal stem cels in threedifferentculture systemswere comparedby oil red O staining and lipogenic geneassay. At 0, 1, 3 and 7 days of adipogenensis, mRNA expressionsof PPARγ, C/EBPα, Adiponectin and Leptin were detected.At7 daysofadipogenensis, protein expressionsof PPARγ and C/EBPβ were detectedby western blot assay,andeffects ofDIMIversusDIMRonphosphorylationofPPARγatSer273were compared.
RESULTS AND CONCLUSION:Findings from oil red O staining andreal-time PCRshowedthat DIMR significantlyinducedadipogenicdifferentiation of bonemarrow mesenchymal stem cels compared with DIM and DIMI at 7 daysofinduction. Western blot showed thattheprotein expressionsof PPARγ and C/EBPβ in the DIMIgroupwere significantly higher than those in the DIMRand DIM at 7days ofinduction. In addition, the ratio ofPPARγphosphorylation atSer273was lowerin the DIMR group thantheDIMI group.To conclude,DIMR has the most potential to induce early adipogenesis ofhumanbone marrow mesenchymal stem cels by weakening the phosphorylationof PPARγ-Ser273.

Objective To investigate the effects of aging on procollagen α polypeptide gene transcription and protein expression in rat vascular smooth muscle cells.Methods Vascular smooth muscle cells from thoracoabdominal aorta in neonate and 9 months old healthy Wistar rats were cultured in vitro.Results Transcription polymerase chain reaction(RT PCR) and Western blotting were used to detect type Ⅰ and Ⅲ pro-collagen α polypeptide mRNA and protein.The RT-PCR displayed that type Ⅰ procollagen α polypeptide mRNA expression had no significant difference between young group and adult group [(76.62±1.05) vs.(78.37±2.42),P＞0.05].Type Ⅲ procollagen α polypeptide mRNA expression was (105.40 ± 2.66) in young group and (123.10 ± 3.81) in adult group(P＞0.05).Type Ⅰ procollagen α polypeptide mRNA expression was (3.13 ±0.54) in young group and (4.63 ± 1.03) in adult group (P=0.05).Type Ⅲ procollagen α polypeptide mRNA expression had no significant difference between the adult and young groups[(6.86 ±0.41) vs.(7.68±0.63),P＞0.05].Type Ⅰ and type Ⅲ procollagen α polypeptide protein expressions were increased significantly in adult group as compared with the young group [(0.10 ± 0.03) vs.(0.06±0.03),(0.58±0.06) vs.(0.40±0.02),both P＜0.05].Conclusions Aging increases the procollagen α polypeptide level in vascular smooth muscle cell,which may involve in the development of vascular remodeling and atherosclerosis.

Objective To discuss the impact of repeated contrast media exposure on renal function in patients who received coronary angiography ( CAG) or percutaneous coronary intervention ( PCI) within 1 week after CTA of coronary ateries. Methods A total of 258 patients who received CAG or PCI after coronary CTA were divided into the study group ( n=132, patients had CAG/PCI within 1 week after CTA) and the control group ( n=126, patients had CAG/PCI 1-2 weeks after CTA). Serum creatinine, cystatin C and estimated GFR were tested before and on day 1, 2 and 3 after procedures. The occurance of contrast-induced nephropathy ( CIN ) was recorded. Resu1ts The baseline clinical characteristics of the patients between the two groups had no significant difference. Preoperative and postoperative serum creatinine, cystatin C and eGFR values on day 1, 2 and 3 had no significant difference between the two groups (all P﹥0. 05). There was no significant difference in the incidence of CIN between two groups (5. 3% in the study group vs. 4. 8% in the control group, P﹥0. 05 ) . Conc1usions It is safe and feasible for patients with eGFR≥60 ml/( min?1. 73 m2 ) to undergo CAG or PCI within 1 week after coronary CTA.

Objective To summarize the experience about the diagnosis and treatment of bladder benign neoplasm in children.Methods A retrospective study was conducted for a total of 15 patients with bladder benign neoplasm from October 2006 to May 2016.There were 10 males and 5 females with a mean age of 8.7 years (ranging 1.1-13.8 years).The clinical manifestations of 15 patients included hematuria in 9 patients,frequent micturition with urgent and painful in 3 patients,dysuria in 1 patient,abdominal pain in 2 patients and headache during voiding in 1 patient.Ultrasound showed solid masses in the bladder with iso-echoic or nonhomogeneously hypoechoic.CT scanning showed regular or irregular mass with some enhancement in the bladder.All cases received tumor complete resection by opening operation and bladders were preserved.Among the 15 cases,neoplasms located in the anterior,posterior and lateral wall of bladder in 9 cases,ureteral orifice in 4 cases and trigone of bladder in 2 cases.The size of tumors ranged from 1.2 to 6.0 cm (mean 3.1 cm).The tumors were unifocal and seemed like papillary or cauliflower.The literatures of benign neoplasm of bladder were reviewed,which focused on the clinic characters,pathological classification and therapeutic method.Results Pathologic type included papilloma in 5 patients,inverted papilloma in 1 patient,inflammatory myofibroblastic tumor in 7 patients,hemangioma in 1 patient,pheochromocytoma in 1 patient.Fifteen patients were followed up for 6 to 36 months,mean 26.7 months.All patients recovered well without relapse or metastasis Conclusions Bladder benign neoplasm in children is rare with many kinds of pathological classification.The major clinical manifestation is gross hematuria while dysuria is unusual.Tumors are fewer in trigone of bladder.The best treatment is to resect the tumor completely with bladder preservation if possible.

Objective To study the expressions of C -met and COX -2 in thyroid papillary cancer ( TPC) and to analyze the relationship between their expressions and clinicopathological features .Methods S-P immunocytochemical technique was used to examine the expressions of C -met and COX-2 in 96 cases of TPC and 46 cases of benign tissues ,and their relationship with clinicopathological features were analyzed .Results The positive rates of C -met and COX -2 in 96 cases of TPC were 92.71% and 71.88%,showing obvious differences between TPC and benign tissues (P<0.001).In TPC there is a statistically significant difference of the expression of C-met in older than45 years and less than 45 years of age(P<0.05).Difference of COX-2 expression in TPC was statistically significant in patients with and without lymph node metastasis ( P<0.05 ) . The positive expression of COX -2 was closely related to the expression of C -met(r=0.451,P<0.05).Con-clusion The abnormal expressions of C -met and COX-2 could be important significance in the growth and in-vasion of TPC,and COX-2 might have some relationship with lymph node metastasis .