Background Oxidative stress is a main cause of age-related macular degeneration (AMD).Lutein has a preventive role for AMD, but its antioxidant mechanism remains unclear.Objective Present study was to investigate the effect of lutein on oxidative stress of Müller cells and its signaling pathway.Methods Human Müller cells (human Müller cell strain) were cultured, and the cells at logarithmimic growth phase were incubated in 96 well plate overnightly.Oxidative stress cell models were established by adding 160 μmol/L H2O2, a median lethal dose for Müller cells.The models were divided into the model control group and 12.5,25.0,50.0 mg/L lutein groups,and the different concentrations of lutein were used to culture the cells for 24 hours, respectively.The routine cultured cells served as the blank control group.Growth of the cells was assayed by MTT method (absorbancy);the reactive oxygen species (ROS) content in the cells was assayed by flow cytometry;the mRNA and protein levels of nuclear factor-E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in the cells were detected by quantitative real-time PCR and Western blot, respectively.Results The inhibitory effects on the cells were gradually enhanced with the increase of H2O2 concentrations,showing a significant difference among the groups (F =43.890,P＜0.01).A significant difference was found in apoptotic rate of the cells among the blank control group,model control group and 12.5,25.0,50.0 mg/L lutein groups (F =346.770, P =0.000) , and the apoptosis rate was significant elevated with the increase of lutein dose (all at P＜0.05).The ROS contents in the cells were 1.92±0.18,64.89±2.86,52.70±2.80,32.61 ±4.20 and 5.68 ± 1.35 in the blank control group, model control group and 12.5,25.0,50.0 mg/L group, respectively, with significant difference among the groups (F =324.900, P =0.000), and the ROS content was gradually reduced as the increase of lutein dose (all at P＜0.05).The relative mRNA and protein expressions of Nrf2 and HO-1 were remarkedly higher in the 12.5,25.0,50.0 mg/L lutein groups than those in the model control group (F =236.960,242.620,186.830,263.120, all at P =0.000) , and no significant difference was seen in the relative expression level of nuclear Nrf2 protein among the groups (F =1.790, P =0.210).Conclusions Lutein can induce the expression of antioxidant enzymes by inducing the expression of nuclear translocation of Nrf2 and consequently inhibit the oxidative stress status.

Objective To evaluate the efficacy, safety and compatibility of a new type of atrial septal defect ( ASD) occluder in atrial spetal defect mini-pigs model.Methods Five Tibet mini-pigs were selected as the ASD models which were established by the combination of atrial septal puncture and balloon dilation.Then the new type occluder was implanted for the therapy of ASD.Transthoracic echocardiography with color Doppler was used in all animals during closure and in follow up examinations.The animals were killed at 3 months after occlusion for electron microscopical observation and microscopic examination.Result The ASD models had been created in five piglets successfully without complication, all of whom were implanted successfully with the new device without shunts.Postmortem and microscopic examination of the 5 specimens 3 months after placement showed complete.Conclusion Transcatheter ASD occlusion with new type ASD occluder is safe,feasible and effective.This occlusion can repair the atrial septal defect successfully.

Objective To amplify the encoding full-length sequence of Tibet mini-pigs myostatin ( MSTN) gene and analysis the sequence by bioinformatics software.Method The RNA of liver tissues from Tibet mini-pig was extracted, and reversely transcribed into cDNA.The gene coding region sequence of myostatin gene was amplified through RT-PCR, and then the purified product of PCR was ligated with a pMD19-T and transferred into the bacterium DH5αfor replication.The positive clones were screened and sequenced. The sequence characters were analyzed by using bioinformatics method and phylogeny evolution tree was constructed with other twelve species.Results The coding redion of MSTN gene was 1128bp, and coded 375 amino acids.The amino acid homology analysis showed that the homology rate of amino acid sequence was 99%.Conclusions Molecular phylogeny evolution indicated that it had a close relation with human, dog, banna minipig, sheep, goat, cattle, horse, chimpanzee, rat, mouse except chick and zebrafish, and the most closely related with banna minipig.

Objective To explore the diagnostic value of cfDNA in pancreatic cancer. Methods A total of 467 patients with pancreatic cancer and 129 healthy controls were enrolled. cfDNA concentration was detected using the QuantiDNA Direct cfDNA Test (DiaCarta) kit. Mann-Whitney U test was used to compare cfDNA concentration between different groups, and Chi-square test was used to analyze the relationship between cfDNA and pathological data of pancreatic cancer. Diagnostic efficacy was evaluated by ROC analysis. Results The cfDNA level of patients with pancreatic cancer was significantly higher than that of healthy controls (20.85 vs. 15.15 ng/ml, P=0.0027). The positive rate of cfDNA in pancreatic cancer was higher than that in healthy controls (77.73% vs. 59.68%). The sensitivity levels of cfDNA, CEA, and CA19-9 as diagnostic markers were 62.65%, 64.04%, and 60.32%, and their specificity levels were 61.05%, 70.53%, and 87.37%, respectively. The sensitivity and specificity of the combined detection were 77.26% and 53.58% respectively. The areas under the curve of cfDNA, CEA, CA199, and combined detection were 0.62, 0.67, 0.74, and 0.67, respectively. Conclusion cfDNA is elevated in patients with pancreatic cancer and may serve as an effective marker for its diagnosis.

OBJECTIVETo test the feasibility of using 1,5-anhydroglucose alcohol (1,5-AG) as a diagnostic indicator of fulminant type 1 diabetes (FT1DM).

METHODSFifteen patients with newly diagnosed FT1DM and 52 with type 2 diabetes (T2DM) were examined for serum biochemistry, glycosylated hemoglobin (HbAlc), and serum 1, 5-AG level.

RESULTSThe patients with FT1DM and T2DM showed significantly different fasting levels of blood glucose (FBG), fructosamine (FMN), creatinine (Cr), urea, HbAlc and serum 1,5-AG (P<0.05). In FT1DM patients, serum 1,5-AG was found to inversely correlate with FBG (r=-0.646, P=0.032) and FMN (r=-0.680, P=0.021), and in T2DM patients, serum 1,5-AG was inversely correlated with FBG (r=-0.407, P=0.001), FMN (r=-0.314, P=0.01) and HbAlc (r=-0.576, P<0.01). Receiver-operating characteristic (ROC) curve analysis showed an area under the curve of serum 1,5-AG of 0.804 with a cutoff value of 67.95, a sensitivity of 82.9% and a specificity of 60% for FT1DM diagnosis.

CONCLUSIONSerum 1, 5-AG can reflect acute blood glucose fluctuation in FT1DM patients and is useful for differential diagnosis of FT1DM when combined with evaluations of the clinical characteristics of the patients and other related indicators.

Blood Glucose ; Diabetes Mellitus, Type 1 ; blood ; diagnosis ; Diabetes Mellitus, Type 2 ; blood ; diagnosis ; Diagnosis, Differential ; Ethanol ; Glycated Hemoglobin A ; chemistry ; Humans ; ROC Curve ; Sensitivity and Specificity

OBJECTIVETo investigate the effect of immunization with prokaryotically expressed recombinant fusion protein of extracellular near-transmembrane domain of Tibet minipig leptin receptor (OBR) on fat deposition in SD rats.

METHODSA pair of specific primers containing BamHI and HindIII restriction enzyme sites was designed to amplify the extracellular near-transmembrane domain (1705-2364 bp) of Tibet minipig OBR gene. After digestion, the amplified fragment was inserted into the plasmid pRSETA between BamHI and HindIII sites. The recombinant plasmid was transformed and expressed in E.coli BL21(DE3) and the product was analyzed by SDS-PAGE and Western blotting. SD rats were immunized with the fusion protein, and the changes in body weight, feed intake, body length, Lee's index, percentage of abdominal fat, liver fat deposition and subcutaneous fat deposition were assessed.

RESULTSThe recombinant fusion protein obtained (about 27.6 kD) was expressed in E.coli induced by IPTG and identified by SDS-PAGE and Western blotting. The rats immunized with the fusion protein showed no significant changes in body weight, body length, Lee's index, percentage of abdominal fat or liver fat deposition as compared with the control rats. Nevertheless, the immunization caused significantly increased feed intake and significantly decreased volume of subcutaneous fat cells.

CONCLUSIONImmunization with the fusion protein of extracellular near-transmembrane domain of Tibet minipig OBR can promote feed intake and suppress subcutaneous fat deposition in SD rats.

Adiposity ; drug effects ; Animals ; Base Sequence ; Female ; Gene Expression ; Genetic Vectors ; Obesity ; Plasmids ; Rats ; Rats, Sprague-Dawley ; Receptors, Leptin ; administration & dosage ; genetics ; Recombinant Fusion Proteins ; administration & dosage ; genetics ; Subcutaneous Fat ; physiology ; Swine ; Swine, Miniature

Objective To investigate and analyze the characteristics of blood physiological and biochemical parameters of Tibetan chickens bred in Guangzhou. Methods Blood samples of Tibetan chickens bred in Guangzhou were collected,and the physiological and biochemical parameters were measured. Results (1)The blood RBC,PLT,PDW, RDW-SD and P-LCR were not significantly different in the males than females(P > 0.05).(2)HCT(P < 0.05), MCHC(P< 0.05),MPV(P< 0.05),HGB(P< 0.01),MCV(P< 0.01)and MCH(P< 0.01)were significantly higher between the males and females.(3)RDW-CV was significantly lower in the blood physiological parameters of males than females.(4)AST,TRIG,ALKP,ALT,Ca,CHOL,CREA,GLU,PHOS and TBIL were not remarkably different in the blood of males than females(P > 0.05).(5)The blood AMYL(P < 0.05)and TP(P < 0.01)were significantly higher in the males than females.(6)The blood ALB(P< 0.01),UREA(P< 0.05), and GLOB(P<0.01)were significantly lower in the males than females. Conclusions The essential data of blood physiological and biochemical indexes of Tibetan chickens bred in Guangzhou are obtained.