Objective　To display efficient folding hDAF on the surface of yeast. Methods　The hDAF open reading frame was cloned by PCR from DAF- pBluescript M13-（Amp＋）plasmid， then subcloned into the yeast surface displayed vector pYD1.The recombinant vector was transformed into yeast cells EBY100.Flow cytometric analysis was carried out to evaluate direct binding of anti-DAF mAbs onto the surface of yeast cells displaying DAF. Results　Three mAbs against DAF different epitopes could bind onto DAF displayed on the surface of yeast.Conclusion　Efficient folding DAF can be displayed on the surface of yeast potentially leading to the development of novel applications involving DAF yeast display.

Objective To explore the relationships between resistin rs1477341 , rs3219175 and type2 diabetes mellitus (T2DM) in Guangdong. Methods 180 patients of diagnosed T2DM in the Second Affiliated Hospital of Guangzhou Medical University from January 2013 to December 2014 were enrolled in the study as the treatment group and another 180 subjects taking health examination as the control group. The genotype distributions of rs1477341 locus A/T, rs3219175 locus A/G were compared and the relationship between the genotypes and T2DM were anlysed by the logistic regression. Results The frequencies of the rs1477341, rs3219175 locus genotype reached the genetic equilibrium, showed their representativeness of a population group. The frequency distribution of rs1477341 genotype was statistically insignificant across the two groups (P > 0.05). The frequency of rs3219175 genotype AA+AG was decreased while genotype GG increased significantly compared to the control group (P < 0.05). The logistic regression results suggest genotype GG may increase disease risk of T2DM which the morbidity rate of AG homozygote was 0.630 times as high as that of GG homozygote (OR =0.630, 95%CI:0.405 ~ 0.980, P < 0.05) and the morbidity rate of GG homozygote was 1.551 times as higher as that of AA+AG homozygote (OR = 1.551, 95%CI:1.016 ~ 2.368, P < 0.05). Conclusions rs1477341 genotype has no correlation with T2DM among the population in Guangdong , but rs3219175 genotype does. GG may have a higher susceptibility to T2DM.

The Monascus mutant with high yield of yellow pigment was obtained by using conventional relevant mutation techniques, e.g., treating with physical mutagens(such as UV light) and chemical sub- stances (such as N-methyl-N'-nitro-N-nitrosoguanidine). The yellow pigment was scanned from 300 nm to 600 nm with UV spectrometer, the maximal absorption was determined at 410 nm. The growth characteristic of Monascus mutant is stable, the yellow pigment value and colour hue in liquid fermentation can reach 100 U/mL and 3.5 respectively. The yellow pigment is stable from pH 3 to pH 8, but the precipitation appeared as the pH of the pigment solution lower than 3.

OBJECTIVETo investigate the proportion between tumors which maintain their telomeres by a mechanism of alternative lengthening of telomeres(ALT) and telomerase-dependent tumors in gastrointestinal malignant tumors, the expression difference of hRad21 between the two groups and the clinicopathological characteristics of ALT tumors were also explored.

METHODSOne hundred and four cases of gastrointestinal malignant tumors were divided into 2 groups: ALT group and telomerase group by detecting telomerase activity using TRAP method. Expression difference of hRad21 was investigated between the two groups. All the patients were followed up and clinicopathological data of these patients were analyzed.

RESULTSOf 104 cases, there were 12 cases in ALT group and 94 cases in telomerase group. Expression of hRad21 in ALT group was higher than that in telomerase group. Tumors in ALT group had a thinner invasion depth (lower T stage) as compared to telomerase group (P=0.021). Other indexes, such as age, gender, tumor size, tumor grade, location of tumor, CEA and CA199, were not significantly different between the two groups. Results of follow-up showed that the survival rate of ALT group was 100% while that of telomerase group was 56% at 30 months postoperatively.

CONCLUSIONSThere are tumors which maintain their telomeres by ALT in gastrointestinal malignant tumors, accounting for 10%-12% of the total tumors. As compared to telomerase group, ALT group presents higher expression of hRad21, thinner tumor invasion depth, and higher survival rate.

Female ; Gastrointestinal Neoplasms ; metabolism ; pathology ; Humans ; Male ; Neoplasm Invasiveness ; Nuclear Proteins ; metabolism ; Phosphoproteins ; metabolism ; Telomerase ; metabolism ; Telomere ; metabolism

Objective:To investigate the promoting effect of Ginsenoside Ro on the differentiation of THP-1-derived dendritic cells (DCs) induced by GM-CSF and IL-4.Methods: Sensitive leukemia-derived DC cell line was screened first.Then,the selected sensitive cell line THP-1 was stimulated to differentiate into DCs by cytokines (GM-CSF and IL-4) and small(5 μmol/L),middle(10 μmol/L),and large (20 μmol/L) dose of Ginsenoside Ro respectively.The expressions of CD1a,MHCⅡ and CD86 of leukemia-derived DCs were detected by flow cytometry.In addition,the transcription levels of CD1a,CD86 and MHCⅡ of leukemia-derived DCs were detected by RT-PCR.ELISA was used to measure the protein levels of TNF-α and IL-6 in the culture supernatant.Results: THP-1 was the sensitive leukemia cell line which could be induced to differentiate into DCs by cytokines.Compared with cytokine stimulation alone,the expression of CD1a,MHCⅡ and CD86 in leukemia-derived cells was significantly increased after the stimulation of Ginsenoside Ro combined with cytokine(P<0.05).The CD1a,CD86 and MHCⅡ mRNA expression was significantly increased after the treatment of Ginsenoside Ro combined with cytokine(P<0.05).Moreover,the protein levels of TNF-α and IL-6 in culture supernatant were significantly increased (P<0.05) after the stimulation of Ginsenoside Ro in combination with cytokines.Conclusion: Ginsenoside Ro can significantly promote the differentiation of leukemia-derived DCs.

Objective To explore the mechanism of deep brain stimulation(DBS)therapy to Parkinson's disease(PD).Methods We produced hemi-parkinsonian rat model with stereotaxically injecting 6-OHDA to right medial forebrain bundle(MFB)and stimulated ipsilateral subthalamu nucleus (STN)with platinum electrodes chronically to investigate the influence of DBS to the expression of Calbindin-28,synaptophysin and tyrosine dioxydase(TH)in Striatum by Western blot.In addition,slices of bilateral PD rats after DBS were stained to observe the expression of Calbindin-28 and synaptophysin in substantia nigra by Immunohistochemistry.Results High frequency stimulation impaired the rotational frequency 31% of unilateral PD rats triggered by apomophine;Long-term DBS increased the expression of TH in innocent striatum of unilateral PD rats 78.6%?9.5%,since the ipsilateral striatum(lesion side) was TH depleted by 6-OHDA insults;Calbindin-28 expression in ipsilateral striatum of hemi-PD rats raised up 75.4%?15.0% and long-term DBS reduced the effect by 43.0%?7.1%,meanwhile Calbindin-28 positive neurons in substantia nigra compacta in sham,PD and DBS rats were 74.5?10.2,75.7?15.6, 33.1?7.8.However,Synaptophysin expression in substantia nigra and striatum kept stable even after long- term DBS.Conclusions Consistent to the treatment to PD patients,DBS to STN alleviated the motor disorder of PD rats,the treatment might be based on regulating the expression of Calbindin-28 and TH.

Objective:TTo evaluate the nutritional status,immune function and clinical effectiveness of early enteral immunonutrition in patients receiving totally endoscopic esophagectomy.Methods:There were 90 patients receiving totally endoscopic esophagectomy enrolled in Department of Thoracic Surgery of The Fourth Hospital of Hebei Medical University between May 2015 and October 2016.The patients were randomly divided into three groups,enteral immunonutrition group (EIN group,n =30),common enteral nutrition group(EN group,n =30)and parenteral nutrition group (PN group,n =30).The levels of nutritional indexes,immune indexes,C-reactive protein (CRP) were measured preoperation and 1day,3days,7days after surgery.We also observed the first postoperative anal exhaust time,infectious complications and the length of hospital stay.Results:There were no significant differences in age,gender,tumor location,clinical stage,CRP,the nutritional indexes and immune indexes between the three groups before operation.On the first day after operation,the nutritional indexes and immune indexes of the three groups were significantly lower than those before operation,then all the indexes began to recover.Otherwise,CRP of the three groups were significantly higher than those before operation,then fell down in the following days,the rate of EIN group and EN group was faster than that of PN group on the day 3 after surgery and the EIN group was the fastest on the day 7 after surgery.There were significant differences in the first postoperative anal exhaust time,pneumonia and the length of hospital stay.And there was no significant difference in the incidence of anastomotic fistula between the three groups(x 2=1.071,P =0.585).Conclusions:Postoperative early enteral immunonutrition can improve the nutritional status and the immune response,promote the rapid recovery in the patients receiving totally endoscopic esophagectomy.

OBJECTIVETo investigate the effect of human caudal-related homeobox 2 (Cdx2) gene expression on human gastric carcinoma cell line MGC-803.

METHODSpCMV-Cdx2-HA eukaryotic expression plasmid was constructed by using DNA recombinant method. The MGC-803 cells were divided into 4 groups: non-transfected, transfected with pCMV-HA, transfected with pCMV-GAPDH-HA, transfected with pCMV-Cdx2-HA. Cdx2 gene and its protein expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot techniques respectively. The effects of Cdx2 overexpression on the growth of MGC-803 cells in vitro was assessed by measuring MTT, flow cytometry and transmission electron microscopy analysis.

RESULTSIn MGC-803 cells transfected with pCMV-Cdx2-HA, cell proliferation was significantly suppressed, the cells was ultrastructurally destroyed, cell cycle progression was blocked in G0/G1 and apoptosis rate was higher (P<0.05) in comparison with non-transfected cells or cells transfected with empty vector.

CONCLUSIONIt indicated that Cdx2 gene can suppress the growth of gastric carcinoma and may save as a novel therapeutic target.

Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Genetic Vectors ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; RNA, Messenger ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

AIMTo study the antitumor effects of an immunoconjugate composed of lidamycin (LDM) and monoclonal antibody 3G11 (3G11-LDM conjugate).

METHODS3G11-LDM conjugate was prepared by using 2-iminothiolane (2-IT) and m-maleimidobenzoyl-N-hydroxy-succimide ester (MBS) as crosslinkers. The molecular weight of the conjugate was measured on non-reduced SDS-PAGE gel. Immunoreactivity of 3G11-LDM conjugate to type IV collagenase or to hepatoma 22 cells was determined by ELISA. The cytotoxicity of the immunoconjugate to hepatoma 22 cells was examined by MTT assay. Antitumor effects of the 3G11-LDM conjugate in vivo were evaluated using subcutaneously transplanted hepatoma 22 tumor model in mice.

RESULTSThe molecular weight of 3G11-LDM conjugate was approximately 160 kDa. 3G11-LDM conjugate retained part of the immunoreactivity of 3G11 to type IV collagenase and hepatoma 22 cells. As compared with free LDM, 3G11-LDM conjugate showed stronger cytotoxicity to hepatoma 22 cells. When administered intravenously (i.v. x 2 on day 1 and 8), 3G11-LDM conjugate, at doses of 0.05 and 0.10 mg.kg-1, inhibited the growth of hepatoma 22 in mice by 87.8% and 97.2% on day 11, respectively, whereas the unconjugated LDM at 0.05 mg.kg-1 inhibited tumor growth by 67.1%. The median survival times for tumor-bearing mice of untreated control, LDM at 0.05 mg.kg-1, 3G11-LDM at 0.05 mg.kg-1, and 3G11-LDM at 0.10 mg.kg-1 were 34, 41.5, 60.5 and 94 d, respectively. Evidently 3G11-LDM was more effective than free LDM in suppressing tumor growth and prolonging the life span of tumor-bearing mice.

CONCLUSION3G11-LDM conjugate shows much stronger antitumor effects than equivalent dose of free LDM and may have promising therapeutic potential in cancer treatment.

Aminoglycosides ; administration & dosage ; therapeutic use ; Animals ; Antibiotics, Antineoplastic ; administration & dosage ; therapeutic use ; Antibodies, Monoclonal ; immunology ; Cell Division ; drug effects ; Disease Models, Animal ; Enediynes ; Female ; Immunoconjugates ; therapeutic use ; Liver Neoplasms, Experimental ; drug therapy ; pathology ; Matrix Metalloproteinase 2 ; immunology ; Matrix Metalloproteinase 9 ; immunology ; Mice ; Neoplasm Transplantation ; Random Allocation ; Tumor Cells, Cultured

OBJECTIVETo study the variation and evolution rules and analyze the genetic characterization of echovirus 30 during an outbreak of aseptic meningitis.

METHODSFor all the echovirus 30 isolates during the outbreaks in Zhangqiu city in 2003, complete VP1 genes were determined. Phylogenetic tree reconstruction and pairwise sequence were determined to compare the homological relationship and evolution rules between Zhangqiu strains and the other reported echovirus 30 in GenBank.

RESULTSPairwise sequence comparisoms in VP1 region indicated that there were 98.9%-99.5% identity of nucleotide acid among Zhangqiu strains, and having the highest homology with Taian strains (98.5%-99.0%) isolated in Shandong province. At the same time, when comparing with Zhejiang and Jiangsu strains, the nucleotide homology showed 98.2%-98.9%, relatively low identity with Taiwan strains (87.4%-87.6%) and French strains (89.1%-89.7%) but only 82.4%-82.8% identity when comparing with echovirus 30 prototype strain Bastianni. Data from phylogenetic tree analyses indicated that all the echovirus 30 correlated with aseptic meningitis in China recently came from the same evolution linkage and formed a monophyletic cluster.

CONCLUSIONIn 2003, there was an circulation of echovirus 30 in Shandong province which causing an endemic of aseptic meningitis in local areas. Data from phylogenetic tree analyses indicated that the antigenic characteristics of echovirus 30 in mainland China might be different from strains early isolated in other continents and formed a new genotype.

China ; epidemiology ; DNA, Viral ; analysis ; Disease Outbreaks ; Enterovirus B, Human ; classification ; genetics ; Enterovirus Infections ; epidemiology ; Genotype ; Humans ; Meningitis, Aseptic ; Phylogeny ; Sequence Analysis, DNA