To develop a multiplex PCR assay to detect Enterohemorrhagic Escherichia coli (EHEC), Vibrio parahaemolyticus (VP), Salmonella spp. and Listeria monocytogenes(LM) in innocuous poulthy and seafood simultaneously, four pairs of primers were designed from hemolysis gene (hlyAB) of EHEC, toxR gene of VP, invA gene of Salmonella spp. and iap gene of LM. They were detected by conventional PCR and multiplex PCR assay. DNA sequencing was used to examine PCR products. This multiplex PCR method was a highly specific, rapid and sensitive method which could simultaneously detect EHEC, VP, Salmonella spp. and LM. The detection level was 10~(3)cfu/mL.

Regulatory T cells are a group of subgroup T cells with special immune regulating function,which participate in keeping immune homeostasis and play important roles in preventing autoimmune diseases.However,regulatory T cells also promote tumor immune evasion.The highly increase of its expression will significantly reduce the smvival rate of cancer patients.Gastric cancer with high mortality is difficult to detect in early stage.China is one of the high-prevalence gastric cancer countrys.This article mainly introduced the sources,classification and immune phenotype of regulatory T cells and its function mechanism to explore its relationship with gastric cancer in order to discover the new strategy of tumor immunotherapy.

BCR/ABL negative classical myeloproliferative neoplasms including three diseases,namely polycythemia veta,essential thrombocythemia and primary myelofibrosis.Progresses have been made in the study of these dieases in children following the development of that in adult,especially the research of molecular pathogenesis.Differences exist in clinical and molecular characteristics between children and adult BCR/ABL negative classical myeloproliferative neoplasms.There are specific characteristics in the diagnoses and treatment of childhood myeloproliferative neoplasms.This paper reviews the characteristics of childhood BCR/ABL negative classical myeloproliferative neoplasms in clinical and molecular features,diagnoses and treatment.

BACKGROUND:At present,mesenchymal stem cells(MSCs) is mainly obtained from stem cells.But the quantity of human bone marrow mesenchymal stem cells(BMSCs) is lower.Fetal MSCs possess weak immunogenicity and good differentiation potential.OBJECTIVE:To compare the biological difference in adult and fetal BMSCs.DESIGN,TIME AND SETTING:The cytology in vitro controlled study was performed at the Central Laboratory,Hospital Affiliated to Medical College,Qingdao University from February to October 2008.MATERIALS:Three samples of adult bone marrow blood were obtained from Hospital Affiliated to Medical College,Qingdao University.Three samples of fetal bone marrows were supplied by Qingdao Municipal Hospital.METHODS:Human and fetal BMSCs were harvested in vitro by the Percoll method,and incubated in LG-DMEM containing 10% volume fraction fetal bovine serum.Till cells were 90% confluent,cells were digested and passaged.The same passage of adult and fetal BMSCs were collected,and treated with KGla suspension and DMEM containing fetal bovine serum at 37 ℃ in 0.05 volume fraction CO2 for 2 hours.Non-adherent cells were removed.At the third passage,BMSCs at a density of 1?106 were incubated.When cells were 60%-80% confluent,cells were incubated in osteoblast medium for 16 days.MAIN OUTCOME MEASURES:Cell morphological changes were measured.Growth curve was detected by MTT assay.Cell adherent rate in vitro was calculated.Alkaline phosphatase staining was used to detect the differentiational results of osteoblasts.RESULTS:Adult and fetal BMSCs showed spindle-shape,but the fetal BMSCs were small.The doubling time of them was 50 hours and 30 hours,respectively.Compared with the adult BMSCs,cell adherent rate was significantly lower in the fetal BMSCs(t=4.22,P

Bone marrow mesenchymal stem cells is a new approach for diabetes treatment, it can transdifferentiate into insulin cells in vitro, that may overcome the shortage of human donor islets, avoid the immunological rejection and moral principle problems which are involved into the research of embryonic stem cells.Studies have shown that insulin gene and its other necessary gene can introduce into primary cells or other somatic stem cells directly in diabetes mellitus patients, such as hepatic cells, intestines epidermis K cells, fibroblast cells and liver tumor cells and so on.Thereby, it can be translated into insulin-secreting cells.At present, the main study direction internationally is how to import insulin gene into target cells effectively, and make it express in a physical model.

Objective To investigate the pharmacokinetics of intravenously infused domestic alfentanil in the patients undergoing general anesthesia.Methods Twelve American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients of both sexes,aged 18-47 yr,weighing 49-86 kg,scheduled for elective subtotal thyroidectomy under general anesthesia,were enrolled in the study.Anesthesia was induced with iv midazolam 0.02 mg/kg,alfentanil 25 μg/kg,propofol 1.5 mg/kg and rocuronium 0.8 mg/kg.The patients were endotracheally intubated and mechanically ventilated.Anesthesia was maintained with inhalation of 0.8％-2.0％ sevoflurane,iv infusion of alfentanil 1 μg · kg-1 · min-1,and intermittent iv boluses of rocuronium 10-20 mg,bispectral index value was maintained at 40-60,and infusion was stopped at 10 min before the end of surgery.Before induction of anesthesia,at 1,3,5,8,10,14,20,35,65,95 and 125 min ofalfentanil infusion,and at 5,15,30,60,120,180,240,300 and 360 min after termination of alfentanil infusion,venous blood samples were collected for determination of the plasma concentration of alfentanil (by high performance liquid chromatography-mass spectrometry/mass spectrometry).DAS 3.0 software was used to analyze the pharmacokinetic parameters of alfentanil.Results The pharmacokinetics of domestic alfentanil was best described by a two-compartment pharmacokinetic model.The distribution half-life was (1.8 ± 0.8) min,and the elimination half-life was (91±22) min.The volume of distribution at steady state was (0.38±0.12) L/kg,and the clearance was (4.3± 1.6) ml · kg-1 · min-1.The elimination of domestic alfentanil followed the first-order kinetics.Conclusion The distribution of intravenously infused domestic alfentanil is fitted to a two-compartment pharmacokinetic model with elimination following the first-order kinetics in the patients undergoing general anesthesia.

Acute kidney injury(AKI)has become the public health problem that harms the human's health, is a common critically illness,characterized by complicated etiology,high prevalence,high risk of combine other organs'injury and high mortality,these become the huge challenge in clinic.Thus,it is valuable in clinic how to improve cog-nition about AKI,to early detection,to early treatment so that postpone pathogenetic condition,increae survival rate and improve prognosis.In recent years,the researchers have processed vast of reseaches about etiology,new biomark-er,risk fator,prognosis,etc.However,these parts are still controversial.This paper summarizes the research progress of AKI.

Objective:To observe the therapeutic effect of donepezil combined nimodipine on vascular dementia. Methods:A total of 45 patients with vascular dementia aging 54~76 years were randomly divided into donepezil +nimodipine group (n= 24)and huperzine + piracetam group (n= 21).Scores of mini-mental state examination (MMSE)and activity of daily living (ADL)were examined in two groups before and after treatment,and therapeu-tic effect was compared between two groups.Results:Compared with before treatment,after treatment,MMSE score [(15.8±6.3)scores vs.(21.6±7.4)scores]significantly rose and ADL score [(50.3±6.4)scores vs.(40.2 ±7.1)scores]significantly reduced in donepezil + nimodipine group;MMSE score [(14.6±6.9)scores vs.(18.5 ±7.2)scores]significantly rose,ADL score [(49.3±7.8)scores vs.(43.5±8.2)scores]significantly reduced in huperzine + piracetam group,P <0.01 all;but there were no significant difference between two group after treat-ment (P =0.08,0.07).Conclusion:Therapeutic effect of donepezil combined nimodipine on vascular dementia is quite to huperzine + piracetam.

Objective To sum up the characteristics of hemorrhagic fever with renal syndrome(HFRS)and acute pancreatitis,to improve the understanding of the cases of atypical HFRS,reduce the misdiagnosis rate and improve the cure rate.Methods 3 cases of HFRS complicated with acute pancreatitis in our hospital from 2011 to 2015 were retrospectively analyzed.Results 3 cases were middle -aged men,with an average age of 46.6,of which two cases occurred in Huangling county in the city,the remaining 1 case occurred in Yichuan county of the city. Occupation was the railway workers,drivers,farmers.Respectively occurred in February,July,December.3 cases of blood routine,urine routine,blood coagulation performance were consistent performance for RBC,WBC,PLT decrease;hematuria,proteinuria;3 cases of plasma amylase were higher than that of more than 3 times the normal value,the serum lipase was higher than the more than 3 times the normal value;hemorrhagic fever antibodies were positive;treatment of 2 cases with hemodialysis treatment,1 case underwent intestinal dialysis treatment,successful treatment of all.Conclusion HFRS with acute pancreatitis patients were less,easy to be misdiagnosed or ignored,easy illness of incur loss through delay and once with acute pancreatitis will add to the original condition,treatment more difficult tend to have higher mortality rate.It should be according to the clinical manifestations,early blood and urine amylase, abdominal B ultrasound and CT examinations of the abdomen as soon as possible,to improve the accuracy of diagnosis, take appropriate treatment measures,especially blood dialysis treatment,can reduce the mortality rate.

Intrauterine infection is one of the most important cause of adverse outcomes in pulmonary diseases among preterm infants.The lung tissues of such infants are still immature, whose contact with pathogens when exposed to intrauterine inflammatory environment may trigger local or even systemic inflammatory response, causing different degrees of harm to fetal or neonatal lung tissue.Such damage could lead to bronchopulmonary dysplasia in preterm infants, neonatal pneumonia, neonatal respiratory distress syndrome, etc., affecting their quality of life.This review is about the research progress and the pathogenesis of intrauterine infection in pulmonary diseases of premature infants.