Objective:To study the effects of the overexpression of hsa-miR-202 on the proliferation and molecular mechanism of lung cancer A549 cells. Methods:A sequence of hsa-miR-202 with ppG/miR/eGFP/Blasitidin pasmid was directionally connected and a eukaryotic expression vector pmiR-202 of the target hsa-miR-202 gene was constructed. pmiR-202 was transtected to A549 cell with Lipofectamine 2000. The WST assay was used to detect the cell proliferation rate, and RT-PCR was used to detect the relative gene expression levels. Western blot analysis was used to detect the IL-10 protein expression levels. The interaction between miR-202 and IL-10 was examined using a luciferase reporter assay. Results:The design from the DNA sequencing results shows that a eukaryot-ic expression vector of miR-202 was successfully constructed. The proliferation inhibition rates of A549 cells by Pmir-202 were 12%, 38%, and 52%. The differences in the treatment group compared with the blank control and negative control groups were statistically significant. The RT-PCR results showed that the relative expression levels of miR-202 increased after transfection with pmiR-202. Over-expression of miR-202 can downregulate the relative gene and protein expression levels of IL-10, and the relative levels were 25%and 0.75, respectively. Compared with the blank control and the negative control groups, the difference was statistically significant (P<0.05). The fluorescent activity was reduced when transfection was performed with miR-202 mimics, and IL-10-3'UTR plasmid was cloned. Conclusion: pmiR-202 effectively inhibited the proliferation of A549 cells and exhibited a time-effect relationship with miR-202 by targeted combination with IL-10 3'UTR to downregulate IL-10 expression in A549 cells.

Objective:To develop and characterize the hydrogel microspheres for embolization.Methods:N-[tris(hydroxymethyl)methyl] acrylamide-gelatin microspheres(TGMs)were prepared by an inverse suspension polymerization approach.Effects of materials on size,water absorption rate and elasticity of the microspheres were investigated.The materials which were included consisted of gelatin in the range of 10.0-100.0 g/L,N-[tris(hydroxymethyl)methyl]acrylamide in the range of 33.3-200 g/L,cross-linking agent N,N'-methylene-bis-acrylamide in the range of 3.3-10.0 g/L,surfactant Span 80 in the range of 0.5-1.8 g/L,and initiator ammonium persulfate in the range of 1.0-5.0 g/L.The appearance of TGMs was observed under microscope.TGMs were analyzed by infrared spectrum(IR).Results:The TGMs were round with smooth surface in view of photograph of microscope.The average diameter of TGMs was increased with the increase of gelatin,monomer or cross-linking agent concentrations but decreased with the increase of surfactant or initiator Concentration.The water adsorption rate of the microspheres was decreased with the increase of gelatin or cross-linking agent concentration but not affected by surfactant concentration.The elasticity of TGMs was increased with the increase of monomer or cross-linking agent concentration,decreased with the increase of gelatin concentration,but not affected by surfactant or initiator concentration.All factors above considered,the final prepared TGMs consisted of 10 g/L gelatin,100.0 g/L monomer,6.7 g/L cross-linking agent,0.9 g/L surfactant,and 3.0 g/L initiator.The average diameter of TGMs obtained was about 700.0 ?m.The water adsorption rate and the elasticity in accordance with the maximum diameter of the microspheres passed through a microcatheter of TGMs were 12.4(g/g),and 1 600.0 ?m,respectively.The results of IR spectra confirmed the polymerization of monomer,resulting in N-[tris(hydroxymethyl)methyl]acrylamide-gelatin microspheres.Conclusion:The developed TGMs seemed to be suitable for clinical embolization according to the surface,average diameter,elastic and hydrophilic property of TGMs.

AIM: To investigate the effect of aminoguanidine (AG) on plasma and renal levels of angiogenesis Ⅱ (AngⅡ), and to identify the relationship of AGEs with AngⅡ in STZ-induced diabetic rats. METHODS: Wistar rats were randomly assigned to three groups. Diabetes was induced, rats were then received AG in treatment group. At the end of 12th week, urine albumin excretion rate (UAER) and calculate creatinine clearance (Ccr) were detected. Periodic acid-Schiff reagent was used to evaluate renal pathology. Plasma and renal AngⅡ were analyzed by radioimmunoassay and immunohistochemistry, respectively. RESULTS: AG treatment significantly prevented the increase in UAER (P<0.01), renal pathology (P<0.01), and level of renal AngⅡ (P<0.01). However, plasma concentration of AngⅡ was higher than that in diabetic rats without AG treatment (P<0.01). CONCLUSION: AG down-regulates renal Ang Ⅱ level, probably by reducing the formation of AGEs, which may be one of the renoprotective factors in diabetic nephropathy. More proofs are needed to identify the result that plasma AngⅡ concentration increases in DMA group.

Since the release rate of protein in hydrogels is directly dependent upon the size of the protein and the hydrogel, how to deliver low molecular weight protein for prolonged periods has always been a problem. In this article, we present a usage of self-assembling peptide (P3) with the RGD epitope on its N terminus. The concentration of the released insulin-like growth factor 1 (IGF-1) was determined by UV-vis spectroscopy and the release kinetics suggested a notable reduction of the IGF-1 release rate. Cell entrapment experiments revealed that IGF-1 delivery by biotinylated nanofibers could promote the proliferation of the mouse chondrogenic ATDC5 cells when compared with cells embedded within nanofibers with untethered IGF-1.
Animals ; Biotin ; Cell Line ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Hydrogels ; chemistry ; Insulin-Like Growth Factor I ; pharmacology ; Mice ; Nanofibers ; Oligopeptides ; chemistry

Objective To investigate certain risk factors for and their impact on abnormal liver function in middle-aged and elderly adults.Methods A case-control study was constructed based on the SAGE cohort of 8642 registered residents aged 50 years or over in Shanghai.Of them,137 individuals with abnormal liver function,defined as aspartate transaminase (AST)＞ 40 U/L or alanine aminotransferase (ALT)＞ 40 U/L,were randomly selected as the observation group,while 411 healthy controls were 3 ∶ 1 matched with the cases in the observation group by gender and age (1 year).Face-to-face administered questionnaires and physical examinations were conducted and serum samples were tested for ALT,AST,glucose (GLU),total cholesterol (TC),triglycreide (TG),hepatitis B surface antigen (HbsAg) and anti-hepatitis C virus antibody (anti-HCV Ab).Chi square test and rank sum test were used for single factor analysis,and logistic regression analysis was used for multiple factors.Results The prevalence of HBsAg positive patients was 12.4 ％ (68/548) Univariate analysis showed that hepatitis virus infection and body mass index (BMI) were associated with abnormal liver function (both P＜0.000).Multivariate logistic regression analysis showed that hepatitis virus infection (OR=1.85,95％ CI:1.04 3.29,P-0.036) and obesity (OR=3.60,95％CI:1.92-6.73,P＜0.001) increased the risk of abnormal liver function,whereas chronic medication (OR=0.51,95％ CI:0.32-0.80,P =0.004) decreased the risk of abnormal liver function.Conclusions Among the study population,hepatitis virus infection and obesity are risk factors for abnormal liver function in middle-aged and elderly people.After adjustment for potential confounders,chronic medication is negatively correlated with abnormal liver function and may be a protective factor for liver function.

Objective To study on virulence characteristics and multilocus sequence type of Vibrio parahaemolyticus isolated from clinic in Beijing Tongren hospital.Methods Total 152 strains of Vibrio parahaemolyticus isolates were collected from diarrheal patients of outpatients in Beijing Tongren Hospital,Capital Medical University from 2009 to 2011.PCR was used to detect hemolysin gene thermo stable direct themolysin gene (tdh),TDH-related hemolysin gene (trh),type Ⅲ secretion system 2 (T3SS2α,T3SS2β)and systematic functional gene (toxRS/new,orf8) for pandemic 03∶ K6 clone and its derivatives.The genetic features of these strains were determined by multilocus sequence typing (MLST).Results 96％ (146/152) VP harbored tdh gene,2％ (3/152) VP harbored trh gene and 100％ (152/152) VP harbored T3SS2 gene.In this study there were 107 pandemic strains (both tdh and toxRS/new positive and trh negative),38 pathogenic strains (tdh positive and/or trh positive) and 6 nonpathogenic strains (both tdh and trh negative).All nonpathogenic strains harbored systematic functional gene (toxRS/new,orf8).Only one pathogenic strains harbored orf8 gene.One clone harbored all virulence gene.In this study there were 16 sequence types,and ST3 is the pandemic sequence type,including 113 strains,and four new sequence types were found.Conclusions In this study more than 90％ Vibrio haemolyticus harbored tdh gene and ST3 was the pandemic sequence type in Beijing.One can get bacterial pathogenic charateristic and population genetics information by virulence gene testing and MLST.

Objective To explore the semitivity of ovarian cancer cell line SKOV3 to paclitaxel,oroteasome inhibitors,bortezomib,and their combination.Methods The methyl thiazolyl tetrazolitim (MTT)assay was applied to examine the cell viability after treatment.The annexin V-propidium iodide apoptosis detection kit was used to determine the apoptosis rate of different groups.Western blot assay was used to evaluate the expression levels of phosphorylated protein kinase B(AKT)and glycogen synthase kinase-3 beta(GSK-3β).Results In MTT assay,the cell viability ratios of the combination group at serial time points from 12,24,36,48 and 72 hours Were(65.2±5.8)%,(58.3±14.4)%,(35.3±5.0)%,(19.2±1.5)%,and(11.4 ±2.5)%,which were significantly lower than those of the paclitaxel group (P<0.05).After arug treatments,apoptosis rates of paclitaxel group,bortezomib group and the combination group were (14.7±0.5)%,(15.1±0.8)%and(20.5±0.7)%respectively.The rate of the combination group was significantly higher than that of non-treated group and paclitaxel group(P<0.05).Western blot assay showed the changes in expression levels of phosphorylated AKT and GSK-3β,which were decreased significantly after paclitaxd and bortezomib combination treatment [(3.2±0.8)%,(19.3±0.4)%;P<0.05].Conclusions The lethal effect of paclitaxel on tumor cells could be increased significantly by its combination with proteasome inhibitors,bertezomib.The AKT/GSK-3β signaling pathway plays an important role in the molecular mechanism of the combination treatment.

Objective:To explore the role of a disintegrin-like and metalloproteinase with throm bospondin typeⅠmotifs-8(ADAMTS8) gene in lung adenocarcinoma and its mechanism.Methods:Selected lung adenocarcinoma data sets GSE75037 and GSE116959 and lung adenocarcinoma transcriptome data and clinical information from the TCGA database; useed R software to analyze differential genes in lung adenocarcinoma tissues, and performed GO enrichment analysis and KEGG signal pathway enrichment for differential genes set analysis; analyzed the expression of ADAMTS8 gene in lung adenocarcinoma and its correlation with patient survival and prognosis; CCK-8 experiment detects the effect of ADAMTS8 overexpression or knockdown on the proliferation of A549 cells; Western blot detected ADAMTS8 overexpression or knockdown effect on Wnt/β-catenin signaling pathway. Results:The combined analysis found that 395 genes were up-regulated in lung adenocarcinoma samples, and 716 genes were down-regulated; ADAMTS8 was under-expressed in lung adenocarcinoma tissues, and its expression level was related to tumor stage; low-expression of ADAMTS8 was associated with shorter survival time, poor prognosis, Cox regression analysis showed that ADAMTS8 can be used as an independent prognostic marker for patients with lung adenocarcinoma; CCK-8 results showed that overexpression of ADAMTS8 can inhibit the proliferation of A549 cells, knocking down ADAMTS8 can promote the proliferation of A549 cells ( P<0.05); Western blot results showed that after overexpression of ADAMTS8, the protein levels of β-catenin, cyclin D1 and c-Myc in cells were significantly reduced ( P<0.05), while knocking down ADAMTS8 could cause the protein levels of β-catenin, cyclin D1 and c-Myc in cells increased significantly ( P<0.05). Conclusions:ADAMTS8 is under-expressed in lung adenocarcinoma and can be used as an independent prognostic marker for patients. Overexpression of ADAMTS8 may inhibit the proliferation of lung adenocarcinoma cells by inhibiting the activity of Wnt/β-catenin signaling pathway.

Objective: To improve the social skills of children with ASD by using Program for the Education and Enrichment of Relational Skills(PEERS ), and to reduce the uncertainty towards ASD and negative emotions for mothers of ASD children. Methods: From September to October 2017, 30 dyads of autistic mother and child were recruited and divided into intervention group and control group (15 mother child dyads each). Based on the content of PEERS social skill, cognitive behavior therapy was delivered in group format, through demonstration, role play and group exercise. At the same time, mother child dyads were trained using parallel social technology. Mothers and children with ASD were investigated using Parents Perception of Uncertainty Scale (PPUS), Patient Health Questionnaire 9 (PHQ-9), Chinese Version of the Beck Depression Inventory II(BDI-Ⅱ-C), Beck Anxiety Inventory (BAI), State Trait Anxiety Inventory(STAI-Form Y), and Autism Behavior Checklist (ABC), Cildhood Autism Rating Scale (CARS), and Social Communication Questionnaire (SCQ). Results: Changes in ASD symptom score in children and emotional score of mothers in the intervention group were less than 0. The total score of mother disease uncertainty(74.93±13.58, 90.40± 9.21 ), ambiguity(31.13±7.07, 38.93±4.73), lack of clarity information(11.93±2.09, 13.80±2.54), unpredictability(9.60±1.99, 12.07±2.89), significantly changed after intervention( t =-3.65, -3.55, -2.20, -2.72, P <0.05). Conclusion Social PEERS group intervention can enhance the social skills of children with ASD, reduce uncertainty of illness among mother of ASD children. Timely disease related information, guidance for mothers to actively participate in child care and training, might help to reduce cognitive bias, depressive and anxiety symptoms among mothers.

Objective:To investigate the changes of serum Golgi protein 73 (GP73) expression and its clinical significance in breast cancer patients before and after radiotherapy.Methods:A total of 135 patients with primary breast cancer who were diagnosed and treated in Qingdao Hospital Affiliated to Shandong First Medical University Hospital from Aug. 2016 to Dec. 2017 were selected and received standard radiotherapy after surgery. Enzyme-linked immunosorbent assay (ELISA) was used to determine the content of GP73 in the serum of patients before and after radiotherapy, and the differences in the expression levels of GP73 before and after radiotherapy in patients with different molecular phenotypes, tumor stages, and pathological types were compared. Association between the expression of serum GP73 and clinical outcome before and after radiotherapy was analyzed.Results:The expression of GP73 in breast cancer patients was (117.69±33.57) mg/L before radiotherapy and (101.88±30.92) mg/L after radiotherapy, and the difference was statistically significant ( t=4.025, P<0.001) . Different molecular phenotypes were stratified and found that the expression of serum GP73 in patients with luminal A, luminal B, HER-2 overexpression, and triple negative after radiotherapy decreased compared with those before radiotherapy, but only the HER-2 overexpression type had statistically significant difference between before and after radiotherapy ( P<0.05) . Stratification according to different tumor stages showed that the expression of serum GP73 in patients with stage I, II, III and IV after radiotherapy was lower than that before radiotherapy, but only the patients with stage II, stage III, and stage IV were compared before and after radiotherapy, and the difference was statistically significant ( P<0.05) . Stratification according to different pathological types showed that the expression of serum GP73 in patients with invasive ductal carcinoma before and after radiotherapy was significantly lower in patients with invasive lobular carcinoma ( P<0.05) . In addition, the 1-year survival rate of the descending group was significantly higher than that of the ascending group, while the local recurrence rate and the occurrence of distant metastasis were significantly lower than those of the ascending group, and the difference was statistically significant ( P<0.05) . Conclusions:The expression level of serum GP73 in breast cancer patients after radiotherapy is significantly higher than that before radiotherapy, and it has a certain relationship with molecular phenotype, tumor stage, and pathological type. At the same time, the increase in serum GP73 expression after radiotherapy may be detrimental to the clinical outcome of patients.