Objectives To develop and evaluate a real-time fluorescent quantitative RT-PCR method for the detection of a novel CXC chemokine macrophage inflammatory protein-2?(MIP-2?)mRNA expression based on TaqMan technique.Method The expression of MIP-2? mRNA was detected by TaqMan real time fluorscet quantitative RT-PCR. Results The dynamic range of the assay varied from 102 to 107 copies. The intra- and inter-assay coefficient variation (CV) were less than

Through combing the objects of the medical digital resources of long-term preservation,analyzing the characteristics of medical digital resources,and discussing challenges coming from medical big data,the paper researches the long-term preservation architecture of medical digital resources primarily from life cycle,standard specification,storage architecture,monitoring management and preservation mode to provide effective guidance for comprehensive long-term preservation construction of medical digital resources.

Objective:To investigate the balance of Th1/Th2 and Tc1/Tc2 in peripheral whole blood from SLE patients and find out the relationship between disease activity and the balance of type 1 and type 2 cytokines.Methods:Peripheral blood cells from SLE patients(active,n=15,inactive,n=20) were stimulated and normal subjects(n=20) with phorbol myristate acetate and ionomycin(PMA/I) for 4 h,then IFN-? and IL-4 in CD3~+CD8~-T cells(Th) and CD3~+CD8~+T cells(Tc) were detected by recently developed intracytoplasmic cytokine-staining techniques,anti-dsDNA antibodies in serum was detected with enzyme linked immunosorbant assay(ELISA) and immunoglobulin in serum and protein in urine were analyzed with rate nephelometry.Results:In SLE patients with active disease,the percent of IFN-?-secreting Th cells significantly increased compared with SLE patients with inactive disease and normal after in vitro PMA/I stimulation,the percent of IFN-?-secreting Tc cels significantly increased compared with normal,but IL-4-secreting Th cells and Tc cells did not vary significantly.In SLE patients with inactive diseases,only the percent of IFN-?-secreting Tc cells significantly increased compared with control.The percent of IFN-?-secreting Th and Tc cells in patients with anti-dsDNA antibody(+) in serum decreased significantly compared with patients with anti-dsDNA antibody(-).Although the percent of IL-4-secreting Th and Tc cells in patients with aberrantly high level of immunoglobulin had no change,but the mean fluorescent intensify(MIF) of IL-4-secreting Th and Tc cells increased significantly compared with patients with normal level of immunoglobulin.There was no correlation between the percent of IFN-? or IL-4 in Th or Tc cells and the content of protein in patients' urine.Conclusion:The balance between type 1 and type 2 cytokine-secreting T cells is disrupted in SLE patients,especially IFN-?-secreting T cells,which is significantly in favour of Th2 cells.The level of IFN-? or IL-4 in Th or Tc cells varied partly with disease activity,which deserved further study.

AIM:Chronic exposure to elevated levels of free fatty acids (FFAs) in type 2 diabetes patients is toxic to pancreatic β-cells.Thioredoxin (Trx)-interacting protein (TXNIP), an endogenous Trx-inhibiting protein, is up-regulated by glucose and is a critical mediator of hyperglycemia-induced β-cell apoptosis in diabetes.However, the effects of FFAs on TXNIP are unknown.In this experiment we observed the effect of palmitate on TXNIP expression in cultured INS-1 islet cells and the pathways involved were analyzed meanwhile.METHODS:After the full basis of preliminary experiment of incubating INS-1 cells with palmitate at different concentrations for different time, INS-1 islet cells were cultured with 0.5 mmol/L palmitate for 24 h.TXNIP expression, cell apoptosis, and expression of transcription factors related to TXNIP transcriptional regulation were determined.RESULTS:Compared with control group, the expression of TXNIP at mRNA and protein levels in palmitate group was significantly up-regulated (P<0.01).Cleaved caspase-3/caspase-3 ratio was increased in palmitate group (P<0.05), and the apoptosis of the INS-1 cells was also significantly increased (P<0.01).Palmitate enhanced the phosphorylation of nuclear factor-κB (NF-κB) (P<0.01), and the NF-κB inhibitors, PDTC and SN50, both blocked the palmitate-induced up-regulation of TXNIP expression.CONCLUSION:Saturated fatty acid palmitate enhances the expression of TXNIP.The mechanism of palmitate-induced TXNIP expression may be associa-ted with the increase in NF-κB phosphorylation.

The gamma-secretase complex mediates the final cleavage of APP to generate the principal component of amyloid plaques in the brains of Alzheimer's disease patients.Four integral membrane proteins (PS,NCT,PEN-2 and APH-1) are essential and sufficient for gamma-secretase activity.To identify the promoter of human nicastrin gene (NCT),its 5' -flanking region has been characterized and a 270 bp fragment containing the TSS (transcription start site) for the promoter activity has been identified.EMSA assays confirmed that all four AP-1 binding sites and two NFAT sites in the NCT promoter region were able to bind relative transcription factors in vitro.Mutations,as well as treatment with PDTC,which adjust the regulatory effect of AP-1 and NFAT,altered NCT promoter activity in both HeLa cells and rat cortical neurons.The results demonstrated that AP-1 and NFAT are involved in the regulation of hNCT transcription and suggest that balanced activation of AP-1 and NFAT ensures a strict temporal and tissue-specific control of NCT transcription.

Many somatic cell typos were obtained by in vitro differentiation or teratoma formation of human embryonic stem ceLls (hESCs). However, it is unclear whether specific cell types can be obtained from hESCs-derived teratoma. It was reported that many kinds of cells, including neural progetfitor/precursor cells (NPCs) and mesenchymal stem cells (MSCs) were isolated efficiently from the teratoma of hESCs through in vitro selection. The teratoma-derived NPCs and MSCs showed specific characteristics of molecular markers similar to the primary NPCs and MSCs. Moreover, these teratoma-induced NPCs and MSCs can be further induced to differentiate into neurons, astrocytes, or adipose and bone cells, reflecting their inherent multi-potencies. Given that teratoma normally contains a mixture of ectoderm, mesodenn, and endoderm lineage cells at different differentiation stage, it was suggested that hESCs-derived teratoma could be an alternative source to generate a variety of uncommitted progenitor cells or terminally differentiated somatic cells, which may be otherwise difficult to obtain through direct in vitro differentiation.

Objective:To observe the efficacy of cytokine-induced killer (CIK) cells on patients with advanced lung cancer. Methods:A total of 90 patients with advanced lung cancer were identified from January 2011 to December 2013. CIK therapy was given to 41 pa-tients in the observation group, whereas the other 49 patients in the control group received the best support treatments without che-motherapy or radiotherapy within one month of inclusion. Following up was conducted for the patients in the two groups, and KPS scores, median survival, and adverse reactions compared. Results:The KPS score in the observation group was higher than that of the control group after treatment (P=0.034). The median survival period of the observation group was eight months, which was one month longer than that of the control group (P=0.044). Major adverse reactions included fever, joint pain, and insomnia, which were recorded 51.22%, 36.58%, and 29.27%of occurrence, respectively. Conclusion:CIK cell therapy improved the quality of life and pro-longed the survival of advanced lung cancer patients with tolerable adverse reactions.

Objective To isolate and identify the potential binding partners of LRRK2,a gene linked to both dominant familial form and sporadic form of Parkinson's disease,thus to further our knowledge of its function.Methods We used a sequence containing full-length of COR domain and part of ROC and MAPKKK domain as bait.The bait amplified by polymerase chain reaction(PCR) was then cloned into a yeast expression plasmid pGBKT7.After being sequenced and analyzed,pGBKT7-bait was transformed into the yeast strain AH109.Western blot was performed to confirm the expression of pGBKT7-bait in AH109 yeast strain.Then human fetal brain cDNA library was trarnsformed into that yeast strain.which could express pGBKT7-bait fusion protein.The yeast strain which contained pGBKT7-bait and human fetal brain cDNA library was plated on quadruple dropout medium (SD/-Trp/-Leu/-His/-Ade)containing X-a-gal.We retested these positive colonies using 2 independent yeast strains AH109 contained pGBKT7-bait or pGBKT7,respectively.At last,these plasmids isolated from these true positive colonies were analyzed by bioinformatics.Results We obtained 9 true positive colonies,these colonies were sequenced, and we performed sequence Blast in GenBank.Three colonies of the 9 positive colonies were not in open reading-frames.Among other 6 colonies,there were known proteins including spermatid perinuclear RNA-binding protein(STRBP)and BCL2-associated athanogene 5 isoform b(BAG5),as well as unknown proteins including tyrosine phosphatase non-receptor type(PTPN23),1(3)mbt-like 3 isoform b(L3 MBTL3),RALY RNA binding protein-like isoform 1(RALYL),and Homo sapiens mRNA for KIAA1783 protein,partial cds(KIAA 1783).Conclusion True positive colonies of LRRK2 are successfully obtained by the yeast 2-hybrid.Our screened proteins may provide a new research clue for revealing biological functions of LRRK2,pathogenesis of Parkinson's disease,and other neurodegerations.

G mutation of SLC26A4, the parents and the second child were carriers of the same mutation, while the fetus had a wild-type form. Conclusion It is feasible to identify deafness related genes by screening for GJB2 and SLC26A4 mutation, thus providing correct prenatal diagnosis and avoiding deaf delivery of baby.

Objective To explore the expression of long noncoding RNA ( lncRNA) RN7SK and its screening value in clinical diagno-sis of gastric cancer. Methods The expression of lncRNA RN7SK in gastric cancer cells and paired gastric cancer tissues were meas-ured by qRT-PCR. The screening efficacy was detected by the receiver operating characteristic ( ROC) curve. Results The expression levels of lncRNA RN7SK in gastric cancer lines SGC-7901(3.91±0.53),MGC-803(3.44±0.29),HGC-27(4.04±0.87)and BGC-823 (4.30±1.13) were markedly higher than that in human gastric mucosal epithelial cell line GES-1, and the difference had statistical sig-nificance(1.02±0.27, t=12.33, 7.48, 7.20 and 4.90, P<0.05). The expression of RN7SK was up-regulated in 85.5％ (47/55) of gastric cancer tissues while down-regulated in the the other 8 tissues ( 14. 5％) . The area under ROC curve ( AUCROC ) of lncRNA RN7SK expression in gastric cancer patients was 0.827 and the 95％ confidence interval (CI) was from 0.746 to 0.907. When the cut-off value was 0.618, the sensitivity and specificity were 0.836 and 0.782 respectively. Conclusion The level of lncRNA RN7SK should be overexpressed in gastric cancer tissues so that it may have high screening efficacy and could be used as a potential molecular marker for the diagnosis of gastric cancer.