Detection of H pylori antibody profile in serum by protein array

AIM： To detect multiple H pylori antibodies in serum samples of individuals who carryHpylori by protein array. METHODS： Recombinant H pylori antigens, urease B subunit （UreB）, vacuolating toxin A （VacA） and cytotoxin associated gene A protein （CagA）, were prepared and immobilized in matrixes on nitrocellulose membrane by robotics to bind the specific immunoglobulin G （IgG） antibodies in serum. Staphylococcus protein A （SPA） labeled by colloid gold was used to integrate the immuno-complex and gave red color signal. The scanner based on charge-coupled device （CCD） could collect the image signal and convert it into digital signal. RESULTS： When human IgG was printed on the membrane in increasing concentrations and incubated with immunogold, a linear dose response curve was obtained and the detection limit for IgG was about 0.025 ng. The cutoff values, which were defined as the mean grey level plus 3 times of standard deviation, were 27.183, 28.546 and 27.402, for anti-UreB IgG, anti- CagA IgG and anti-VacA IgG, respectively, as 400 human serum samples with negative H pylori antibodies were detected by the protein array. When 180 serum samples from patients in hospital were employed for detection of IgG against UreB, CagA and VacA, the sensitivity of the protein array was 93.4%, 95.4%, 96.0%, and the specificity was 94.8%, 94.4% and 97.5%, respectively, as compared with the results obtained by ELISA. The assay also showed high reproducibility, uniformity and stability, and the results were available within 30 min. CONCLUSION： The protein array is a very practical method for rapid detection of multiple antibodies in serum samples. It is especially useful for large scale epidemiological investigation of the infection of Hpylori.

Effect of Shenfu injection on microcirculation effect index in early-and middle-stage of cardiogenic shock rats

OBJECTIVE Shenfu injection(SFI)is an effective treatment of cardiogenic shock,the pathology of the central link was microcirculation disturbance.However,whether the microcirculation status of the early-and mid-stage of cardiogenic shock has any difference is unclear.This study aimed to observe the effect of SFI on the microcirculatory disturbance in mesentery for early-and mid-stage of cardiogenic shock rat.METHODS The early-and mid-stage model of cardiogenic shock was established by ligating the ending or root of left anterior descending coronary arteries(LADCA).The rats were randomly divided into 9 groups,ie control group,early-stage model group,mid-stage model group,3 early medicated groups and 3 mid medicated groups(the dosage was 1,3.33,10 mL·kg^(-1) SFI for cardiogenic shock rats of early-and mid-stage,respectively).Parameters in mesenteric microcirculation,such as velocity of RBCs in venules,diameters of venules,the count of leukocyte adhesion and vascular permeability which calculated by FITC-dextran leakage were observed through an GeneandiM2 inverted intravital microscope and high-speed video camera system.RESULTS The cardiogenic shock induced by ligating the LADCA resulted in a number of responses in microcirculation,including a significant increase in the counts of adhesive leukocytes,narrowing of the vascular diameter,decrease in the velocity of RBCs and dextran efflux.All of the above parameters for early-stage cardiogenic shock rats were attenuated by the treatment with SFI,especially the dosage of 10 mL·kg^(-1).While SFI had no apparent time-effect on the vascular diameter and vascular permeability in mesentery for mid-stage cardiogenic shock rats.CONCLUSION The microcirculation status of the early-and mid-stage of cardiogenic shock rats were quite different.The efficacy of early treatment with SFI was more obvious than the mid administration,which could provide experimental and theoretical basis for the patients with cardiogenic shock in an earlier time.

Association between red blood cell increase and gut microbiota induced by high-fat diet

OBJECTIVE To investigate the underlyingmechanism on the association of red blood cell and gut microbiota in rats induced by High-Fat Diet(HFD).METHODS A total of 36 male SpragueDawley rats(180±20 g) were randomly divided into two groups.The control group(n=10) was given a normal chow diet(10% calories of fat),and the High-fat diet group(n=26) was given a HFD(60% calo.ries of fat).We recorded body weight,length and detected serum glucose,serum lipids and insulin ev.ery two weeks.The fresh arterial blood was collected during the experiments and blood gases were measured immediately(Radiometer Medical ApS,Denmark).Thehematocrit(Hct) and partial pressure of oxygen(pO_2) were detected by the sensor cassette,following themanufacturer′s instructions.The de.tection method was conductivity measurements and current method,respectively.The feces from ce.cum were analyzed by 16 S rRNA gene high-throughput sequencing(Illumina Miseq,USA).RESULTS According to the insulin resistance(IR),body weight and body length,the model group was divided into two small groups.(1) IR group,in which IR,body weight and body length were higher than the control group(P<0.05).(2) un-IR group,body weight and body length were higher than the control group(P<0.05),but the IR was not significantly different.In addition,the levels of hematocrit(Hct),checktotalhe.moglobin(ctHb) and check total blood oxygen content(ctO_2) showed significantly increased in the IR group when compared with the control group(P<0.05),however,the pO_2 was not statistically signifi.cant.Furthermore,we identified that the genus Lactobacillus was moderate positive correlation with Hct,ctHb and ctO_2(P<0.05).Compared with the control group,the relative abundance of the Lactoba.cillus was significantly lower in IR group(P<0.05).CONCLUSION The high-fat diet induced rats′ local tissue hypoxia under the red blood cell increasing,oxygen partial pressure constant and the reduction of Lactobacillus′ abundance might be caused by aerobic oxidation and glycolysis inhibition in the meantime.

Effect and mechanism of Huanglianjiedu decoction on high fat-induced atherosclerosisrats and metabolomics study by ultra-performance liquid chromatography-mass spectrometry

OBJECTIVE To explore the biomarkers and molecular mechanism of Huanglianjiedu decoction(HJD) on high fat diet-induced experimental atherosclerosis in rats.METHODS SD male rats were randomly dividedinto five groups(n=8):normal control group,model group,and three dosage groups(1.5,3 and 6 g crude drug per kilogram of body weight).Atherosclerosis was induced by the combination of regular intraperitoneal injection of vitamin D3 and high fat diet for 8 weeks.HJD was administered by oral gavage from the third week once per day and until the end of the study.After the final administration,the blood samples were collected for biochemical analyses [total cholesterol(TC),triglycerides(TG),highdensity lipoprotein(HDL-C),low-density cholesterol(LDL-C)] and blood gas analyses(PaO_2,PaCO_2,pH,ctHb,etc);the abdominal aorta sections were stained with hematoxylin and eosin for histopathology;the liver homogenate were determined for MDA,SOD,OX-LDL,MCP-1 and VCAM-1.The plasma samples were detected using ultraper formance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS).The data of endogenous compounds were preliminarily preprocessed by software Progenesis QI and then analyzed by multivari.ate statistical analysis software EZinfo 2.0 to screen the distinguished biomarkers and the metabolic pathways were analyzed through website http://www.metaboanalyst.ca/.RESULTS Compared with the normal control group,the content of TC,TG,LDL-C,PaCO_2,MDA,Ox-LDL,MCP-1 and VCAM-1 were significantly increased and HDL-C,PaO_2,ctHb and SOD decreased in the atherosclerosis rats.HJD could significantly attenuated the high fat-induced atherosclerosis pathological injury and the abovementioned indexes(P<0.05).The five groups could be clearly distinguished using the metabolomics method.The administration groups profile exhibited an apparent returning trend from that of the model group and that of the normal control group.Twenty-one endogenous metabolites has been significantly changed in atherosclerosis rats.HJD could remarkably up-regulate 5-L-glutamyl-taurine,L-beta-aspartylL-glutamic acid,histidinyl-hydroxyproline,tryptophyl-alanine,4′-O-methyl-(-)-epicatechin,and downregulate protoporphyrin IX,azelaic acid,lacto-N-triaose,cinnamoylglycine and 9′-carboxy-alpha-tocotri.enol.CONCLUSION The beneficial effect of HJD in high fat-induced atherosclerosis rats may be due to anti-oxidant and anti-inflammatory.And it is suggested that HJD may affect the model rats through tryptophan metabolism,taurine and hypotaurine metabolism,histidine metabolism,lysine degradation and porphyrin and chlorophyll metabolism pathway.

Effect of laser shock peening on combined low- and high-cycle fatigue life of casting and forging turbine blades

Laser shock peening （LSP） is a novel effective surface treatment method to improve the fatigue performance of turbine blades. To study the effect of LSP on combined low- and high-cycle fatigue （CCF） life of turbine blades, the CCF tests were conducted at elevated temperatures on two types of full-scale turbine blades, which were made of K403 by casting and GH4133B by forging. Probabilistic analysis was conducted to find out the effect of LSP on fatigue life of those two kinds of blades. The results indicated that LSP extended the CCF life of both casting blades and forging blades obviously, and the effect of LSP on casting blades was more evident; besides, a threshold vibration stress existed for both casting blades and forging blades, and the CCF life tended to be extended by LSP only when the vibration stress was below the threshold vibra- tion stress. Further study of fractography was also conducted, indicating that due to the presence of compressive residual stress and refined grains induced by LSP, the crack initiation sources in LSP blades were obviously less, and the life of LSP blades was also longer; since the compressive residual stress was released by plastic deformation, LSP had no effect or adverse effect on CCF life of blade when the vibration stress of blade was above the threshold vibration stress.