Objective To construcr a phage display library of human single-chain Fv antibodies from peripheral blood lymphocytes of people with positive serum antibody against hepatitis B virus core protein. Methods The heavy chain and light chain variable region gene of human immunoglobulin were amplified individually by RT-PCR from peripheral blood Lymphocytes mRNA of patients with positive serum hepatitis B virus core protein antibody and randomly combined through a DNA linker encoding the peptide (Gly 4Ser) 3 to construct single-chain variable fragments (ScFv)gene. In the presence of helper phage M13KO7, the ScFv were displayed on the surface of recombinant phages. Results Repertoire antibody library was obtained with 10 6 clonies resistant to ampicillin and kanamycin. Conclusions A typical phage display library of human single-chain Fv antibodies has been constructed Using RT-polymerase chain reaction (RT-PCR) and phage display technique, human single-chain Fv antibodies can be screened from this library.

The aim of the study is to investigate the effect of nardosinone (Nar) on neuronal injury induced by oxygen-glucose deprivation (OGD) in primary cortical cultures isolated from embryos at gestational day 14. MTT method was used to determine the dosage regimen of Nar in primary neuronal cultures and observe the influence of Nar on the neurons suffering OGD; Western blotting analysis was used to detect expressions of protein kinase A (PKA), Ras related protein 1 (Rap1), mitogen-activated protein kinase kinase 1 (MEK1) and phospho-extracellular signal-regulated kinase 1/2 (p-ERK1/2) of OGD-injured or uninjured primary cultured neurons after Nar treatment. Results showed that Nar (50 and 100 micromol x L(-1)) improved the cell viability during OGD damage (P < 0.01) and increased the expression of PKA, Rap1, MEK1 and p-ERK1/2 in injured neurons. Additionally, elevations of PKA, Rapl, MEK1 and p-ERK1/2 in uninjured neurons were caused by Nar (50, 100 and 200 micromol x L(-1)) with a dose-dependent tenclency as well (P < 0.01). In conclusion, Nar could protect against the neuronal injury exposed to OGD, which may be relevant to the promotion of PKA and ERK signaling pathway.

Objective To explore the predicting value of peripheral blood CD20+ CD24hi CD38hi regulatory B cells (Bregs) on the prognosis of elderly patients with sepsis.Methods A prospective study was conducted. Septic patients aged > 65 years old, compliance with diagnostic criteria for Sepsis-3, admitted to emergency and emergency intensive care unit (ICU) of Shanghai East Hospital of Tongji University from April 2016 to February 2017 were enrolled. Procalcitonin (PCT), C-reaction protein (CRP) and lactate (Lac) were routinely measured. According to the worst clinical index value within 24 hours, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and sequential organ failure assessment (SOFA) score were recorded. The concentrations of peripheral blood CD20+ CD24hi CD38hi Bregs were measured by flow cytometry at 1, 3 and 7 days after diagnosed in elderly patients. All patients with sepsis were followed up for 28 days and then divided into death group and survival group according to 28-day outcome. The difference of clinic data and Bregs were compared between the two groups. The significant different factors of elderly sepsis patients were analyzed by binary logistic regression analysis. The correlation between Bregs level and other indicators was analyzed by Spearman correlation. The receiver operating characteristic curve (ROC) was used to evaluate the prognosis value of Bregs in elderly patients with sepsis.Results Fifty-eight patients were enrolled in the study, with 38 male and 20 female; age of (79.91±7.97) years; 32 in sepsis group, 26 in septic shock group; 35 deaths, 28-day mortality rate was 60.3%. APACHE Ⅱ score and SOFA score in death group exhibited much higher than that in survival group (APACHE Ⅱ: 18.14±4.52 vs. 14.91±3.56, SOFA: 8.80±4.56 vs. 6.35±3.00, bothP < 0.05), the Bregs was significantly decreased at 1, 3 and 7 days in death group [cells/μL: 0.70 (0.20, 1.40) vs. 1.50 (0.70, 2.20), 0.54 (0.20, 1.00) vs. 1.42 (1.10, 2.12), 0.25 (0.10, 0.50) vs. 0.80 (0.50, 1.00), allP < 0.05]. Correlation analysis showed that the concentrations of peripheral blood Bregs at 1 day in elderly patients with sepsis was negatively correlated with APACHE Ⅱ score (r = -0.351,P = 0.007), and it was not correlated with PCT, CRP, Lac or SOFA score. It was shown by binary logistic regression that Bregs [odds ratio (OR) = 1.865,P = 0.028] and APACHE Ⅱ score (OR = 0.853,P = 0.026) were independent risk factors for elderly sepsis outcome. It was shown by ROC curve analysis that the prognostic value of the levels of Bregs at 1, 3, 7 days and APACHE Ⅱ score were higher in the elderly patients with sepsis, and the area under ROC curve (AUC) and 95% confidence interval (95%CI) were 0.842 (0.647-0.954), 0.770 (0.564-0.911), 0.888 (0.703-0.977), 0.855 (0.661-0.961), respectively, allP < 0.01. The 7-day Bregs was most powerful to predict outcome, when the cut-off value was 0.50 cells/μL, the sensitivity was 72.73% and specificity was 86.67%. Conclusions The level of peripheral blood CD20+ CD24hi CD38hi Bregs could predict the clinical outcome of elderly patients with sepsis.

Objective To provide a better reference for the quality control of Nujin Pian by establishing the method for the determination of paeonol in Nujin Pian by HPLC. Methods HPLC was used Agilent XDB C18 column (4.6 mm×150 mm, 4.6 μm), mobile phase was composed of methanol-water (45∶55). The flow rate was 1.0 mL/min with detection wavelength at 274 nm and column temperature at 40 ℃. Results The regression equation of paeonol was Y=7×107X-16 965, r 2=0.999 8 (n=6). The linear range of paeonol was 0.003 8-0.38 μg. The average recovery rate was 99.7%, and RSD was 1.56% (n=6). Conclusion The method is simple, fast and accurate with good reproducibility. It can provide reference for the quality control of Nujin Pian.

Objective To understand the influence of personal feature on nurses' professional identity.Methods Nurses of 6 military hospitals were assessed and analyzed by general data questionnaire,Nursing Professional Identity Scale.Results Nurses' professional identity in military hospital was on the moderate level.Nursing age,department,monthly income,position,gender entered into the regression equation of nurses' occupation,which affected positively nursing professional identity.Conclusions The military nursing managers should raise the income of nurses appropriately,especially pay attention to training of professional identity of seniority nurses and local recruitment nurses,and enhance master concept in order to increase their understanding and recognition of their occupation.

Aim To investigate the effects of maternal chronic aluminum exposure on memorial behaviour and hippocampal intracellular Ca2+ concentration ([Ca2+]i)on their offspring after the induction of LTP(long-term potentiation). Methods Adult Wistar rats (150~200 g) were exposed to aluminum by drinking distilled water, the concentration of AlCl3 is 0.015 mol?L-1(2 g?L-1) and 0.03 mol?L-1(4 g?L-1) aluminum chloride (AlCl3) solution, respectively, for 30 days prior to mating and during the whole gestation and suckling period. Their offspring were distributed into three experimental groups: control group; two exposed groups (represented by 0.2%-Al and 0.4%-Al ) administrated aluminum exposure ended at postnatal day 21. The brain tissue and blood aluminum levels were measured by Atomic absorption spectrometry (AAS). Memorial ability of the offspring was tested by Step down test.[Ca2+]i was measured by the technique of Fura-2/AM calcium ions fluorescence indicator. Results The mean aluminum content in blood and brain tissue was significantly higher than the control group(P0.05), but was significantly decreased in 0.4%-Al exposed group(P

AIM: To construct an eukaryotic expression vector of human single-chain variable fragment ~against hepatitis B virus core protein (anti-HBc ScFv) and detect its expression in HepG2 cells. METHODS: Anti-HBc ScFv genes were amplified from the plasmids abstracted from positive clone and inserted into pEGFP-c1 vector that contained green fluorescent protein gene. The recombinant plasmids were transfected into HepG2 cells, and resistant clones were obtained by G418 selection. The expression of the gene of fusion protein was determined by fluorescent invert microscope and ELISA. RESULTS: Recombinant plasmids were successfully constructed. The plasmid transfected HepG2 cells were obtained by G418 selection. Specific fluorescence was observed in HepG2 cells 48 hours after transfection. ELISA analysis confirmed the expression of anti-HBc ScFv in the cells. CONCLUSION: The construction of human anti-HBc ScFv eukaryotic expression vector and its expression in HepG2 cells lay the foundation for advanced research of intracellular anti-HBc ScFv.

AIM: To Screen and identify human single-chain variable fragment (ScFv) specific to hepatitis B virus core protein and determine its gene sequence. METHODS: The recombinant phages were panned by HBcAg coated in a 96-pore plate and 48 clones were identified specific to HBc after three rounds of panning. The specificity of ScFv from the positive clone was determined by ELISA. Then, the soluble ScFv was expressed in E.coli. HB2151 and secreted in the supernatant. Subsequently, SDS-PAGE and dot blot were performed to identify the ScFv in the supernatant and cell lysate. The gene of ScFv specific to hepatitis B virus core protein was sequenced. RESULTS: The ScFv screened from phage antibodies has a specific combination character with hepatitis B virus core antigen. Soluble ScFv was confirmed to express in E.coli. HB2151 and secrete in the supernatant. The sequence of ScFv gene conformed to that of heavy chain and kappa chain of human immunoglubulin. CONCLUSION: Human ScFv specific to hepatitis B virus core protein has been identified by means of the phage display technology, and its gene sequence has been determined.

Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; mortality ; Humans ; Infant ; Male

Objective To observe characteristics of the activity in resting brain of asymptomatic reflux esophagitis (RE) patients,and explore the role of default network in the pathogenesis of asymptomatic RE.Methods Functional magnetic resonance imaging (fMRI) was performed to scan the brains of 15 asymptomatic RE patients,15 symptomatic RE patients and 15 healthy volunteers under no-mission-stimulation state.With fractional amplitude of low frequency fluctuation (fALFF),characteristics of the strengthened and weakened regions of asymptomatic RE patients,symptomatic RE patients and healthy individuals were compared and analyzed.SPM 5 was applied for data preprocessing.Single-sample t test was performed to observe the noticeable active regions of each group under resting state,and then double-sample t test was used to compare the characteristics of active regions of the entire brain between asymptomatic RE patients and symptomatic RE patients or healthy individuals.Results Under resting state,the active brain regions of asymptomatic RE patients,symptomatic RE patients and healthy individuals were similar.The activity of only small part of brain regions in visceral sensory processing relevant networks changed.There was no significant difference in the active regions of default network under resting state between asymptomatic RE patients and healthy individuals.Compared with symptomatic RE patients,fALEF decreased at right central sulcus cover,right superior temporal gyrus,right insula,right transverse temporal gyrus,right postcentral gyrus,superior temporal gyrus and temporal pole (X=50,Y=-6,Z=12,t=2.7874,P＜0.01).Conclusion The difference in characteristics of activity in default network under resting state may play an important role in the pathogenesis of asymptomatic RE.