We obtained recombinant ferritin from Dermatophagoides f arinae ,and analyzed the characterization of the pro‐tein .A pair of primers was designed according to the known sequence of ferritin gene .The live mites identified and cultured lo‐cally were picked and the total RNA was extracted .The ferritin gene fragment was amplified by RT‐PCR ,and cloned into pET32a vector ,and then transferred into E .coli Top10 .The target gene obtained from the recombinant plasmid by digestion with Bam HⅠand Hind Ⅲ was connected to the prokaryotic expression vector pET‐32a .The expressed recombinant plasmid containing ferritin gene was constructed by cloning target gene into pET‐32a and transferred into E .coli Bl21 (DE3) .The ex‐pressed recombinant protein was analyzed by SDS‐PAGE ,and was purified by immobilized metal ion affinity chromatography (IMAC) .The ferritin expressed by dust mite was analyzed by the method of bioinformatics .The recombinant plasmid pET32a‐ferritin was constructed .SDS‐PAGE showed a correct molecular weight of the recombinant ferritin protein .After purification by affinity chromatography ,the protein showed only one strip on SDS‐PAGE gel .SDS‐PAGE showed a band at 20 kD .Dust mite ferritin contains 8 serine kinase ,7 threonine kinase ,7 tyrosine kinase ,and 0 histidine kinase phosphorylation sites .Hy‐drophilic region is larger than the hydrophobic region and it is an unstable protein .In conclusion ,the ferritin gene has been cloned and expressed .The purified ferritin has high purity . The study provides a basis for further study of composition and physicochemical properties of house dust mite allergen .

To study the prognosis-related regulation mechanism of miR-452-5p and its influence on the proliferation and migration of hepatocellular carcinoma (HCC) cells, liver hepatocellular carcinoma (LIHC) dataset in The Cancer Genome Atlas (TCGA) was used to validate the differential expression of miR-452-5p and perform the Kaplan-Meier analysis of overall survival (OS).Target genes of miR-452-5p from TargetscanHuman and miRDB databases were predictived; and differentially expressed genes(DEGs) and weighted gene co-expression network analysis (WCGNA) were completed with GSE14520.Lipofectmine-2000 was used to transfect miR-452-5p mimics, mimics negative control, miR-452-5p inhibitor and inhibitor negative control into Huh7 cells,respectively.The mRNA and protein expression level of RORα in 4 groups were determined by RT-qPCR and Western blot.CCK-8 assay and Transwell assay were conducted to testify the capabilities of proliferation and migration.The regulation between miR-452-5p and RORα was confirmed by the dual luciferase reporter assay.After analysis in the TCGA-LIHC dataset, miR-452-5p had higher expression in HCC tissue than that in normal tissue, which was also associated with a shorter OS.RORα and LAMC1 were discriminated by intersecting of DEGs, WGCNA module genes, and predictive target genes.Survival analysis exhibited that dysregulation of RORα was significantly related to the OS.Overexpression of miR-452-5p in HCC cells suppressed the expression of RORα both in mRNA and protein, and also enhanced the viability and migration of HCC cells.The results of the dual luciferase reporter assay showed that miR-452-5p targeted 3′UTR of RORα.Up-regulated miR-452-5p inhibited the expression of RORα, facilitated the proliferation and migration of HCC cells, and promoted the progression and poor prognosis of HCC.

MEK is a canonical effector of mutant KRAS; however, MEK inhibitors fail to yield satisfactory clinical outcomes in KRAS-mutant cancers. Here, we identified mitochondrial oxidative phosphorylation (OXPHOS) induction as a profound metabolic alteration to confer KRAS-mutant non-small cell lung cancer (NSCLC) resistance to the clinical MEK inhibitor trametinib. Metabolic flux analysis demonstrated that pyruvate metabolism and fatty acid oxidation were markedly enhanced and coordinately powered the OXPHOS system in resistant cells after trametinib treatment, satisfying their energy demand and protecting them from apoptosis. As molecular events in this process, the pyruvate dehydrogenase complex (PDHc) and carnitine palmitoyl transferase IA (CPTIA), two rate-limiting enzymes that control the metabolic flux of pyruvate and palmitic acid to mitochondrial respiration were activated through phosphorylation and transcriptional regulation. Importantly, the co-administration of trametinib and IACS-010759, a clinical mitochondrial complex I inhibitor that blocks OXPHOS, significantly impeded tumor growth and prolonged mouse survival. Overall, our findings reveal that MEK inhibitor therapy creates a metabolic vulnerability in the mitochondria and further develop an effective combinatorial strategy to circumvent MEK inhibitors resistance in KRAS-driven NSCLC.

BACKGROUNDCoronary heart disease (CHD) is the most common type of heart disease and cause of heart attacks. This study investigated the epidemiological characteristics of CHD and its risk factors in Jiaozhou, Shandong province, to ultimately find a way of reducing the prevalence of cardiovascular disease, and to provide a theoretical basis for establishing a cardiovascular disease management path under the regional medical collaborative mechanism.

METHODSA questionnaire survey was performed including 1 952 people aged 35 years or older who were questioned by means of stratified, cluster, proportional sampling to investigate the prevalence of CHD and its risk factors. The data were inputted into SPSS11.0 statistical software for processing and analysis. We advised the local medical institutions to establish health files for the residents with CHD and risk factors. They were followed up regularly. Their risk factors and life-style were monitored, and advice was given as to proper medications. Green channels were established, and the patients were transmitted in a timely manner to superior hospitals for better treatment if the necessary treatments were not available in the local hospitals. The control of risk factors was observed after the follow-up for half a year.

RESULTSIn Jiaozhou, the rates of coronary artery disease, hypertension, diabetes, hyperlipidemia and overweight were 8.15%, 28.54%, 11.43%, 35.46%, and 18.70% respectively. The rates of hypertension, diabetes, hyperlipidemia and overweight were higher than the data published in "The report of Chinese cardiovascular disease 2012"; which are 24%, 9.7%, 18.6%, and 9.7%, respectively. The control of risk factors improved significantly after the guidance of the residents lifestyle and medication for six months.

CONCLUSIONSThe high prevalence of coronary artery disease in Jiaozhou is closely related to age, gender, diet structure, family history of cardiovascular disease, hypertension, diabetes, hyperlipidemia, overweight, and unhealthy lifestyle. Under the regional medical coordination mechanism, the collaborative management of cardiovascular disease can provide new management concepts for the areas short of medical resources, so as to reduce the prevalence of cardiovascular disease.

Adult ; Aged ; China ; epidemiology ; Coronary Disease ; epidemiology ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Risk Factors