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.

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.

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: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 summarize early experience in combined thoraco-laparoscopic esophagectomy with two-field lymph node dissection for esophageal carcinoma.

METHODSA total of 150 patients with thoracic esophageal cancer who underwent combined thoracoscopic and laparoscopic esophagectomy in Union Hospital, Fujan Medical University, were enrolled in this study.

RESULTSLocations of the tumors included upper esophagus (n=14), middle esophagus (n=95), and lower esophagus (n=41). Pathological type showed squamous cell cancer (n=142) and other types of cancer (n=8). There was no intraoperative death. Conversion to open thoracotomy was required in 6 patients and conversion to open laparotomy in 2 patients. The average total operative time was( 258±45) min. The average operative thoracoscopic time was (140±33) min. The average time for gastric mobilization and neck esophagogastric anastomosis was (119±28) min. The average blood loss during the procedure was (207±130) ml. The average number of harvested lymph node with the specimen was 23.3±8.2. The tumor staging included stage I (n=39), II (n=58) and III (n=53). Postoperative complications occurred in 48(32%) patients including pneumonia (n=17), recurrent laryngeal injury (n=13), anastomotic leak (n=9), arrhythmias (n=9), chyle chest (n=5), delayed gastric emptying (n=5), ileus (n=2), volvulus (n=1), and thrombocytopenia (n=1). Two patients died postoperatively due to respiratory failure resulting from pneumonia.

CONCLUSIONThoraco-laparoscopic two-field lymph node dissection of esophageal cancer is a feasible minimally invasive approach.

Adult ; Aged ; Aged, 80 and over ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; methods ; Lymph Node Excision ; methods ; Male ; Middle Aged ; Retrospective Studies ; Thoracoscopy ; methods

OBJECTIVETo develop a TaqMan real-time PCR for the detection of Aeromonas hydrophila.

METHODSThe conserved region of major adhesion gene of Aeromonas hydrophila (aha) was used to design primers and TaqMan probe. A total of six concentration gradients for forward and reverse primers ranging from 200 -700 nmol/L were chosen, and four concentration gradients for probe ranging from 100 - 400 nmol/L were chosen. And then the concentration of primers and probe were optimized by ANOVA of completely randomized design respectively. The specificity of the established method was evaluated by using bacteria as contrast, including 45 strains Vibrio cholerae, 20 strains Vibrio parahaemolyticus, 10 strains Vibrio fluvialis, 4 strains Vibrio mimicus, 5 strains Vibrio vulnificus, 1 strain Vibrio alginolyticus, 1 strain Vibrio furnissii, 5 strains Salmonella, 10 strains Shigella and 2 strains Plesiomonas shigelloides. The sensitivity, bacterial sensitivity and DNA sensitivity included,were evaluated. The stool of healthy people was contaminated by Aeromonas hydrophila artificially, and the ability of the established TaqMan real-time PCR system for detection of Aeromonas hydrophila was also evaluated.

RESULTSThe cycle threshold (Ct) value deserved from 6 groups of primers concentration gradient was (x +/- s):20.69 +/- 0.33, 20.72 +/- 0.21, 20.81 +/- 0.12, 20.74 +/- 0.12, 20.51 +/- 0.16 and 20.69 +/- 0.11, respectively, and the concentration of forward primer and reverse primer was determined to be 200 nmol/L (F = 1.33, P = 0.28). The Ct value deserved from 4 groups of probe concentration gradient was (x +/- s):20.56 +/- 0.08, 20.82 +/- 0.05, 20.82 +/- 0.11 and 20.93 +/- 0.09, respectively, and the concentration of probe was determined to be 100 nmol/L (F = 5.26, P = 0.01). The bacterial sensitivity and DNA sensitivity were 80 CFU/ml and 100 fg/microl respectively, and the sensitivity to detect Aeromonas hydrophila from stool was 8 x 10(3) CFU/ml, which might be 8 CFU/ml after 8 hours' enrichment. No amplification was observed in the templates of other bacterial.

CONCLUSIONThe TaqMan real-time PCR method targeting the aha gene of Aeromonas hydrophila had a high sensitivity and specificity and might be used to detect Aeromonas hydrophila from pure bacterial and stool rapidly.

Aeromonas hydrophila ; genetics ; DNA Primers ; Genes, Bacterial ; Molecular Probes ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Species Specificity

OBJECTIVETo observe effects of electroacupuncture (EA) of low frequency on heroin-seeking behavior and FosB protein expression in relative brain regions so as to explore the mechanism of EA.

METHODSRat model of relapsing into heroin was established with progressive fixed ratio program, and model rats were randomly divided into 3 groups: a "Sanyinjiao" needle-retention control group, a low frequency and weak EA group, and a low frequency and strong EA group. Heroin-seeking behavior was elicited by conditional clue and small dose of heroin; FosB protein expression was investigated with immunohistochemical technique.

RESULTSAfter treatment, the heroin-seeking behavior induced by conditional clue decreased in the needle-retention control group and the weak EA group, and the heroin-seeking behavior induced by small dose of heroin in the weak EA group significantly reduced as compared with the control group, and FosB protein expression in the nucleus accumbens septi, globus pallidus, basolateral amygdaloid nucleus significantly decreased in the weak EA group, and did not significantly change in the strong EA group; the activity induced by heroin increased as compared with those in the control group and the weak EA group.

CONCLUSIONEA of low frequency and low intensity can cure the heroin-seeking behavior, which is correlated with regulating nervous adaptation of nucleus accumbens septi, basolateral amygdaloid nucleus, etc..

Amygdala ; chemistry ; Animals ; Electroacupuncture ; methods ; Globus Pallidus ; chemistry ; Heroin Dependence ; therapy ; Immunohistochemistry ; Male ; Nucleus Accumbens ; chemistry ; Proto-Oncogene Proteins c-fos ; analysis ; Rats ; Rats, Sprague-Dawley