Objective To evaluate the immune response triggered by an in-house constructed hu-man metapneumovirus multi-epitope antigen ( MEA) in a mouse model .Methods Female SPF BALB/c mice at age 4-6 weeks were used in the study and divided into 7 groups.Mice in the five groups including MEA+oligodeoxynucleotides containing CpG motifs ( CpG ODN) intraperitoneal injection ( i.p.) treatment group, MEA+Alum i.p.treatment group, MEA+Alum+CpG ODN i.p.treatment group, MEA+CpG ODN intranasal (i.n.) treatment group and MEA+Alum+CpG ODN i.n.treatment group were immunized three times on days 0, 14 and 21, and those in the other experimental group were immunized intramuscularly with MEA+Quickantibody5W on days 0 and 21.A control group without treatment was set up accordingly .All mice were sacrificed two weeks after the last immunization .Antibodies including IgG , IgG1, IgG2a and IgA in serum samples were detected by ELISA .MTS assay was performed to analyze the proliferation of lympho-cytes.The cytotoxicity of cytotoxic T lymphocytes (CTL) was measured by LDH assay.Flow cytometry was used to detect T lymphocyte subsets .The cytokines secreted by T helper cells ( Th1 and Th2) were analyzed with Bio-Rad Liquid Chips.Results High titers of IgG, IgG1 and IgG2a antibodies were produced in MEA treated mice except for those in intranasal treatment groups .Serum samples from three groups including the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups were positive for IgA antibody .The highest titer of IgA antibody was detected in mice from the MEA+Alum+CpG ODN i.p.treatment group, which was 2.15×103.Compared with the control group, significantly enhanced proliferation of lymphocytes was observed in the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups (P<0.05).Enhanced cytotoxic activities of CTL were observed in mice with ip.and i.m.treatments as compared with those in control group (P<0.05).The levels of CD4+/CD8+T cells were slightly increased in mice from the MEA+CpG ODN i.p., MEA+Alum+CpG ODN i.p. and MEA+Quickantibody5W i.m.treatment groups as compared with those in control group (P<0.05).In-creased secretion of IL-2, IFN-γand Th2-type cytokines including IL-4, IL-5 and IL-10 were detected in mice from the MEA+CpG ODN i.p.treatment group.The MEA+Alum i.p.treated mice showed a slightly increased secretion of IFN-γand significantly increased secretions of IL-4, IL-5 and IL-10.Significantly in-creased secretions of IFN-γ, IL-4, IL-5 and IL-10 were detected in mice from the MEA+Alum+CpG ODN i.p.treatment group.Significantly increased secretions of IFN-γ, IL-5, IL-10 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were detected in mice from the MEA+Quickantibody5W i.m.treatment group.Conclusion MEA together with different adjuvants could stimulate high titers of specific antibodies , increase the proliferation of lymphocytes and enhance the cytotoxic activities of CTL .CpG ODN could bal-ance the Th1/Th2-mediated immune responses , and the balance could be enhanced when using CpG ODN in combination with Alum .A similar effect could be achieved by using the commercial adjuvant Quickanti -body5w.This study has paved the way for further investigation on the development of hMPV epitope vaccines and diagnostic reagents for hMPV as well as the epidemiological study of hMPV .

Objective To study proteome variation between uropathogenic E. coil (UPEC)132, UPEC J96 and non-uropathogenic E. coli K-12 MG1655. Methods Two-dimensional gel electrophoresis(2-DE) was applied to compare the differential expression proteins between UPEC 132, UPEC J96 and non-uro-pathogenic E.coli K-12 MG1655. The differential expression proteins were digested in gel by enzyme. The mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from peptide mass fingerprinting (PMF) were re-searched using the internet available database. Results The number of protein spots recognized from UPEC 132 was 466±11, significantly more than that of E. coli K-12 MG1655 (338±15) and UPEC J96 (382±12); there were 298 protein spots shared by the three E.coli strains, 56 protein spots shared by two UPEC strains, and 89 protein spots characterized by UPEC 132. Twenty-two differential expression or significantly increased expression protein spots, involved in virulence factors, metabolism and transportation, regulation of protein synthesis, biological oxidation and unknown functions, were successfully identified by MALDI-TOF-MS. Condusion The proteome from UPEC 132 and non-uropathogenic E. coli K-12 MG1655, or UPEC 132 and UPEC J96 was differentially expressed. It will provide important information on the pathogen-esis of UPEC 132.

To explore the prevalence of hepatitis B virus (HBV) in multiple myeloma (MM) patients, as well as to compare the clinical characteristics and outcome between HBV infected and non-HBV infected patients. Methods:The serology markers of HBV were detected in 363 MM patients and 11227 cases of healthy controls through chemiluminescence. HBV-DNA was measured via real-time quantitative chain reaction. Results:Sixteen out of 363 MM patients (4.4%) were HBsAg-positive, showing significant difference with healthy controls (2.4%). No statistically significant differences were observed in terms of sex, age, type of monoclonal (M) protein, International Staging System (ISS) stage, stem cell transplantation, and risk stratification between HBsAg-positive and HBsAg-negative patients. No significant effect of HBV infection was found on the OS of MM patients. HBV reactivation was observed in two HBsAg-positive MM patients who were treated with combination chemotherapy, including bortezomib and dexamethasone. The replication of HBV could be inhibited by anti-HBV drugs. Conclusion:A higher prevalence of HBV infection was revealed in MM patients. Close monitoring of HBV replication should be conducted in MM patients with HBV infection before and during the courses of chemotherapy.

Objective To explore the relationship between the volume and function of the heart and the pathogenesis of vasovagol syncope (VVS) through the detection of the left atrial volume index(LAVI).Methods The 68 cases in the observation group were diagnosed as VVS and hospitalized in the First Hospital of Jilin University from Jan.1 to Dec.31 in 2012.The 60 cases in the control group were children and adolescents receiving healthy physical examinations during the same period.All the patients were given the examination of heart color Doppler ultrasound,head up tilt test(HUT),body height,body mass,chest X-ray and accounted the LAVI and cardiothoracic ratio was accounted.Results The average age in the observation group and the control group was(12.19 ± 2.01) and(12.15 ± 2.00) years old,respectively.And there was no statistically significant difference in age between these two groups (t =0.10,P ＞0.05).There were 23 boys and 45 girls in the observation group,and 31 boys and 29 girls in the control group.There was statistically significant difference in the ratio of gender composition between these two groups (x2 =4.16,P ＜ 0.05).The LAVI values in these two groups were (21.23 ± 2.04) mL/m2 and (23.45 ± 3.01) mL/m2,respectively.There was statistically significant difference between two groups(t =4.29,P ＜ 0.05).The LAVI values in VVS mixed inhibition (VVS-MI),VVS vascular inhibition (VVS-VI) and VVS cardiac inhibition (VVS-CI) were (21.41 ± 2.98) mL/m2,(21.06 ± 2.59) mL/m2 and(21.23 ± 3.22) mL/m2,respectively.There were statistically significant differences between VVS-MI or VVS-VI and the control groups(t =3.27,3.36,all P ＜ 0.05),but there was no statistically significant difference between VVS-CI and control groups(t =1.61,P ＞ 0.05).The cardiothoracic ratio were 0.43 ± 0.07 and 0.46 ± 0.06 in the observation group and the control group,respectively,and there was statistically significant difference between these two groups(t =3.05,P ＜0.05).Conclusions The pathogenesis of VVS is related to the size and function of left heart.The children and adolescents with smaller LAVI and cardiothoracic ratio are more susceptible to VVS.

Objective To evaluate the treatment efficacy of leukocyte reduction in hyperleukocytic acute myeloid leukemia (HAML) patients with leukostasis grading score (LGS).Methods The data of 54 HAML patients were analyzed retrospectively.The relationship between LGS and leukocyte stasis symptoms or early mortality was observed, and the impact of leukapheresis on LGS was analyzed.Results Among 54 patients with HAML, there were 1 case of M1, 16 cases of M2, 10 cases of M4, 20 cases of M5 and 7 cases of unclassified AML.Based on clinical symptoms and LGS system, 3 cases were LGS 0, 15 cases LGS 1, 17 cases LGS 2, and 19 cases LGS 3.In patients with LGS ≤ 2, the rates of type Ⅰ respiratory failure, central nevers system (CNS) symptoms and early mortality caused by leukostasis were significantly lower than those in patients with LGS 3 (P ＜ 0.05).The LGS of HAML patients was reduced by leukocyte reduction therapy (P ＜ 0.000 1).The LGS of HAML patients treated by leukapheresis and low dose chemotherapy was improved significantly than that of patients treated without leukapheresis (P =0.008).Among 37 cases receiving induction chemotherapy, 20 cases reached complete remission (CR) after the first cycle of induction chemotherapy.CR rate of patients with LGS ≤ 2 was no significantly different compared with that of patients with LGS 3 (P =0.703).Conclusions LGS can be used to evaluate the degree and the improvement status of leukostasis after treatment in HAML patients.The early death often occurres in patients with high LGS.Leukapheresis combined with low-dose chemotherapy can effectively improve the LGS of HAML patients.

Objective To observe the differences of metabolite ratios among mild cognitive impairment (MCI),Alzheimer’s disease (AD) and normal cognitive state (NC)patients in the hippocampus.Methods According to the clinical features,patients were divided into three groups:MCI group (n=30),AD group (n=28)and NC group (n=30).All the patients were examined by 1 H MRS and compared the ratios of NAA/Cr,Ins/Cr,NAA/Ins,Cho/Cr of both the left and right side of the hippocampus.Results The NAA/Cr in MCI group and AD group were much lower than that in NC group (P <0.05).The Ins/Cr and NAA/Ins in MCI group and AD group showed significant differences compared with NC group (P <0.05).On Ins/Cr and NAA/Ins of the left side,there were significant differences among three groups (P <0.05).Conclusion 1 H MRS as a non-invasive diagnostic technique has higher sensitivity in the early diagnosis and differential diagnosis between MCI and AD patients.

Objective To compare the efficacy of Graves disease animal models induced by thyroid stimulating hormone receptor (TSHR) plasmid DNA (pcDNA3.1-TSHR) and by TSHR A subunit recombinant adenovirus(Ad-TSHR289),and to investigate the influence of duration for preparing animal model induced by Ad-TSHR289 on Graves hyperthyroidism and its related indices.Methods The plasmid group and the adenovirus group were set up respectively.The plasmid group:21 female BALB/c mice were randomly divided into model group (n =12) and control group (n =9).The model group were injected intradermally with pcDNA3.1-TSHR 50 μg,once every 3 weeks,totally 3 times.Then 4 weeks after the last immunization,the mice were euthanized to obtain blood for testing TSHR antibody (TRAb),total T4,and thyroid tissue for histological examination.The controls were injected with the same dose of pcDNA3.1 in the same way.The adenovirus group:52 female BALB/c mice were divided into 10-week model group (n =8),14-week model group (n =10) and 18-week model group (n =8),and the respective controls (n =8,n =10,n =8) were set up.All model groups were injected intramuscularly with Ad-TSHR289,three times at three weekly intervals.Then the mice were euthanized at 4,8 and 12 weeks to test TRAb,total T4 level and to observe the change of thyroid histology.The controls were treated with the same dose of Ad-lacz in the same way.Another 8 mice were scheduled to test the dynamic variation of TRAb before and after the 3 times immunization.Results In the plasmid model group,only two of 12 mice developed weak antibody responses against TSHR,and no elevated total T4 level and no hyperplasia changes of thyroid were observed.In the 10-week model group,all mice had high level TRAb [(807.65 ± 136.33)U/L,Six-eighths mice had hyperthyroidism exhibited hyperplasia changes.In the 14-week model group,the TRAb level [(650.12 ± 192.88) U/L]and the incidence of hyperthyroidism (3/10) were lower than those in 10-week group.Histologically,the degree of thyroid hyperplasia lightened to a small extent,but its positive rate did not decline.In the 18-week model group,only 2 of 8 mice displayed slightly elevated TRAb level,and no mice showed increased total T4 level.Additionally,thyroid tissues of 2 mice were mildly abnormal.Compared with the model groups at different time,the change of antibody levels of the mice for TRAb dynamic observation exhibited the similar trend.Conclusions Being good at repeatability and high incidence of hyperthyroidism,the animal model of Graves disease induced by Ad-TSHR289 is still an ideal research tool presently.The duration of model ean be maintained 18 weeks,and 10 weeks is the best period to snstain characteristic of Graves disease.

Objective To explore the expression and clinical significance of proliferation associated antigen Ki-67 in acute myeloid leukemia (AML). Methods A total of 45 AML patients (including 36 newly diagnosed AML patients and 9 recurrent AML patients) and 20 healthy volunteers (healthy group) were enrolled from October 2012 to January 2016 in Department of Hematology in Fuxing Hospital. The expression of Ki-67 in bone marrow blast cells were detected by flow cytometry (FCM). The relation between Ki-67 level and clinical characteristics, and the prognostic significance of Ki-67 were studied. Results The positive rate of Ki-67 in newly diagnosed AML, recurrent AML patients and healthy controls were (10.38±8.41)%, (20.99± 11.49) % and (40.77±11.97) %, respectively. The positive rate of Ki-67 in newly diagnosed AML patients or recurrent AML patients were significantly lower than that in healthy controls (all P<0.05). The positive rate of Ki-67 in newly diagnosed AML patients was significantly lower than that in recurrent AML patients (P=0.006). The level of Ki-67 in newly diagnosed AML patients did not significantly correlated with age, FAB subtype, white blood cell count, a history of myelodysplastic syndrome (MDS), level of lactate dehydrogenase (LDH), proportion of blats cells, NPM1 gene mutation, FLT3-internal tandem duplication (ITD) gene mutation, chromosome karyotype and response to induction therapy (all P>0.05). There was no significant difference of overall survival between high Ki-67 expression group and low Ki-67 expression group in newly diagnosed AML patients [(780±110) d vs. (788±118) d, P=0.927]. Conclusions The proliferation of blast cells in AML patients is lower than that in healthy controls. Detecting the level of Ki-67 may provide a reference for choosing the cell cycle specific chemotherapy drugs in clinical practice. Monitoring Ki-67 during AML process contributes to monitoring disease progression and predicting recurrence.

Clinical data of 12 cases of Trousseau syndrome with cerebral infarction as initial presentation admitted in the neurology department of the Fourth Affiliated Hospital of Hebei Medical University from December 2011 to December 2017 were retrospectively analyzed.Of the 12 patients,4 cases had risk factors for cerebral infarction and 8 ones had no risk factors.There were 2 patients with 1 lesion and 10 patients with two or more lesions in brain imaging.The infarction lesions of 9 patients were located in 2 or more arterial blood supply areas.Ten patients showed an elevated plasma D-dimer level,5 had elevated fibrinogen level,7 showed increased blood platelet count and 8 had increased homocysteine level.Ten cases were confirmed by pathology,2 cases by clinic and imaging diagnosis.The study suggests that multiple lesions with several cerebral arteries involved,high plasma D-dimer and fibrinogen levels may be the clinical characteristics of Trousseau syndrome with initial presentation as acute ischemic stroke and lacking of risk factors.The hypercoagulation state may be the important pathogenesis of this disorder.

Objective To construct and rescue recombinant influenza virus strains expressing hu-man metapneumovirus ( hMPV) epitopes. -ethods B cell, CTL and Th epitopes predicted by bioinformat-ics software were coupled together in different combinations. These different array genes were inserted into the NS1 gene of influenza virus strain A/PR/8/34 ( PR8 ) , respectively. Recombinant PR8 influenza virus vectors expressing different hMPV antigenic epitopes were rescued by reverse genetics using eight-plasmid system. Sequencing analysis was conducted to verify whether the rescued viruses carried the chimeric hMPV epitopes. Hemagglutination ( HA) titers, half tissue culture infection dose ( TCID50 ) and growth curves were detected. Results Interval sequences GPGPG and KK were introduced into hMPV epitope combinations to construct multi-epitope antigens (MEA). These MEA were inserted into the PR8 NS gene, respectively. Using 8 plasmid system, three recombinant influenza virus strains were rescued successfully. After cultured for three passages in Madin-Darby canine kidney ( MDCK) cells and one in eggs, these three recombinant strains could proliferate steadily. Whole genome sequencing verified that the three recombinant strains car-ried the chimeric MEA sequences, named as rFLU/hMPV/B, rFLU/hMPV/CTL-Th and rFLU/hMPV/B-Th. HA titers of the recombinant strains were 128, 128 and 256 using turkey erythrocyte, respectively. Their TCID50 were 107. 0/ml, 106. 8/ml and 107. 0/ml, respectively. Growth curve tests also verified that the recombinant strains could proliferate steadily in MDCK cells. Conclusions Three recombinant influenza vi-rus vector strains carrying the B cell, CTL and Th epitopes of hMPV were rescued successfully. This study lays the foundation for further evaluation of the immune effects of these recombinant viruses and their poten-tial application value in vaccine development.