A total of 13 mumps virus strains were isolated using Vero/Slam cell line from the patients' throat swabs and urines during mumps outbreaks and sporadics in liaoning Province from 2008 to 2011. Fragments of 316 nucleotides containing SH genes from 13 mumps virus isolates were amplified by RT-PCR, the PCR products were sequenced and analyzed. Based on the 316 nucleotides of SH gene, the phylogenetic trees were constructed with WHO mumps reference strains downloaded from GenBank and 13 mumps virus strains. It showed that the 12 mumps virus strains in 2008-2011 belonged to F genotype, nucleotide acids and amino acids similarities were 94.9%-100.0% and 83.3%-100.0%. Compared to the F reference strains, nucleotide acids and amino acids similarities were 92.4%-97.2% and 96.5% 84.2%, indicating that liaoning mumps strains had large genotype variation. Furthermore, compared to other genotype mumps strains, 6 nucleotides mutations (C(Nt65), C(Nt105), G(Nt137), C(Nt192), C(Nt239), G(NT262)) in SH gene were shared in all of liaoning F genotype mumps viruses. However these mutations were not found in other genotypes of mumps viruses. Conserved amino acids in SH protein of some liaoning mumps strains had changed (S to P at position 2 P to L at position 6 T to N at positon 23 L to P/R at position 48). The genotype specific triplet IML (at position 28-30) was changed to TMP in strain 2008-001-007 2011-015 s, nucleotide acids and amino acids similarities of strain 2008-001-07 were 87.5% and 79.8%, compared to F reference strains respectively, and 96.8% and 97.4% respectively compared to the G reference strains, in dicating strain 2008-001-07 belonged to G genotype,which was first found in mainland China.
Amino Acid Sequence ; China ; Genetic Variation ; Genotype ; Humans ; Molecular Sequence Data ; Mumps ; epidemiology ; virology ; Mumps virus ; chemistry ; classification ; genetics ; isolation & purification ; Mutation ; Phylogeny ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics ; metabolism

Objective:To study the morphology of middle cerebral artery ( MCA ) M1 segment .Me-thods:We selected the MRA data of 794 MCA (400 of the left side and 394 of the right side ) from Ja-nuary 1, 2011 to June 30, 2011 consecutively and analyzed the morphology of the MCA M 1 segment in axial, anteroposterior and lateral view , measured the length of the M1 segment, and analyzed the simila-rity of the left and right side M1 segment morphology .Results:In axial, anteroposterior and lateral view , the MCA M1 segment showed C-shape >L-shape >S-shape .In axial view , it was about 373 ( 47%) M1 segment performance for the C-shape, of which 340 (42.8%) M1 segments showed bowing to the dorsal side, only 33 (4.2%) M1 segments showed bowing to the ventral side .In anteroposterior view, it was about 322 (40.6%) M1 segments of the performance of the C-shape, of which 262(33.0%) M1 segments showed a bowing to the superior , 60 (7.6%) showed bowing to the inferior .The similarity of the left and right MCA M1 segments was 27.2% (114/419) in axial view and 42.7% (179/419) in anteroposterior view.It was more similar in anteroposterior view than in axial view .Along with the increase of age, in the axial view, L-shape converted to C-shape very obviously, but only mildly elevated in S-shape .In anteroposterior view , the L-shape converted to the C-shape or S-shape along with the increase of age.Conclusion:The different morphology of MCA M1 segment in axial and anteroposterior view may be involved in the development of intracranial atherosclerosis .

A flavivirus, Culex flavivirus, was first isolated from Chinese mosquitoes with high sequences similarities to those of flaviviruses found in America and Japan. In this study, a total of 48 pools of field-collected mosquitoes were sampled from Dandong of Liaoning Province, China during July to September of 2011. Six isolated viruses showing cytopathic effect (CPE) in C6/C36 cells were tested by reverse transcription polymerase chain reaction(RT-PCR)using Flavivirus genus--specific primers and Culex flavivirus-specific primers and the positive PCR-product was sequenced and compared with the sequences of 10 isolates from GenBank. Phylogenetic tree of NS5 and enevelop genes of flavivirus were constructed. The GenBank accession numbers of NS5 gene were JQ409188, JQ409186, JQ409187, JQ409191, JQ409189 and JQ409190. The GenBank accession numbers of envelope gene were JQ065883, JQ065882, JQ065881, JQ065879,JQ065877 and JQ065878.
Animals ; Base Sequence ; Cell Line ; China ; Culex ; classification ; virology ; Flavivirus ; classification ; genetics ; isolation & purification ; Insect Vectors ; virology ; Molecular Sequence Data ; Phylogeny ; Viral Proteins ; genetics

Objective To evaluate the homogeneity of the proficiency test samples to verify whether it meets the requirements of the comparison in international reference laboratory.Methods According to the Guidance on Evaluating the Homogeneity and Stability of Samples Used for Proficiency Testing (CNAS-GL03),14 biochemical indexes including ALT,AST,ALP,GGT,CK,LDH,TP,T-Bil,Urea,Cr,UA,Glu,TG and TC in the past three years (from 2014 to 2016)were tested by the Roche detection system Modular P800 Biochemical analyzer.The mean ((x)),standard deviation (s)and coefficient of variation (CV)of the samples were calculated.One-way analysis of variance (ANOVA)was performed and the guideline of Ss ≤0.3σ was used to evaluate the between-bottle differences.Results The results showed that the CVs of AST in RELA 2014A and B were higher than 2.0％.The CVs of CK were over 2％ in all tests except for RELA 2016B.The results of ANOVA for RELA samples demonstrated that the F value of CK was over the critical value 4.39,which was statistically significant (P ＜ 0.05).The F values of the ALT and T-Bil in 2015B and the Cr in 2014A were also over 4.39 (P ＜ 0.05) respectively,while the F values of other measurements were less than the critical value of F,indicating there was no statistical significance (P ＞ 0.05).The CK measurement data Ss ＞ 0.3σ in all the samples by the guideline of Ss ≤ 0.3σ,suggesting that there was a between-bottle difference in CK.The other indexes were Ss ≤ 0.3σ,showing no between-bottle difference in those items.Conclusion There were significant differences between the bottles of the CK item in the past three years,and the homogeneity of all the other items in the samples could meet the requirements of Proficiency Testing for the international reference laboratory.

In the present study, extracellular recording was used to examine the neuronal activity of the basolateral nucleus (BL) of the amygdala and the effects of systemic administration of the selective 5-HT(1A) receptor antagonist WAY-100635 on the neuronal activity in the normal rats and rats with 6-hydroxydopamine (6-OHDA)-produced lesions in the substantia nigra pars compacta (SNc). The results showed that the firing rates of BL projection neurons and interneurons were (0.39±0.04) Hz and (0.83±0.16) Hz in the normal rats, and (0.32±0.04) Hz and (0.53±0.12) Hz in 6-OHDA-lesioned rats. There was no significant difference in the firing rates of BL projection neurons and interneurons between the normal and 6-OHDA-lesioned rats. In the normal rats, all BL projection neurons fired in burst; 94% of BL interneurons fired in burst and 6% fired irregularly. In 6-OHDA-lesioned rats, 85% of BL projection neurons displayed a burst firing pattern and 15% fired irregularly; 86% of BL interneurons had a burst firing pattern and 14% fired irregularly. The distribution of firing patterns of projection neurons and interneurons in the BL in 6-OHDA-lesioned rats did not differ from that in the normal rats. Systemic administration of WAY-100635 at 0.1 mg/kg body weight did not change the mean firing rates of projection neurons and interneurons in the BL in both normal and 6-OHDA-lesioned rats. However, a higher dose of WAY-100635 at 0.5 mg/kg body weight significantly decreased the mean firing rate of BL projection neurons from (0.43±0.07) to (0.15±0.02) Hz in the normal rats (P<0.01), but significantly increased the activity of BL projection neurons in 6-OHDA-lesioned rats from (0.37±0.08) to (0.69±0.18) Hz (P<0.004). The mean firing rates of BL interneurons in the normal and 6-OHDA-lesioned rats did not change after administration of a higher dose of WAY-100635 at 0.5 mg/kg body weight. These results demonstrate that the activity of BL neurons after substantia nigra dopaminergic lesion in the SNc is regulated by activation of intrinsic and extrinsic inputs, and that 5-HT(1A) receptors significantly contribute to the regulation of the activity of BL projection neurons in both normal and 6-OHDA-lesioned rats. Furthermore, WAY-100635 induced an increase in the mean firing rate of projection neurons in the BL in 6-OHDA-lesioned rats, suggesting that 5-HT(1A) receptor is likely to play a role in generating affective symptoms in Parkinson's disease.
Action Potentials ; Amygdala ; drug effects ; Animals ; Neurons ; drug effects ; Oxidopamine ; adverse effects ; Piperazines ; pharmacology ; Pyridines ; pharmacology ; Rats ; Receptor, Serotonin, 5-HT1A ; Serotonin 5-HT1 Receptor Antagonists ; pharmacology ; Substantia Nigra ; pathology

OBJECTIVETo study the effect of dexamethasone on airway morphology and on the expression of angiopoietin-1 (Ang-1) and its tyrosine kinase receptor Tie-2 in the airway of asthmatic rats.

METHODSForty-five Sprague-Dawley rats were randomly divided into control, asthmatic, and dexamethasone-treated asthmatic groups. Asthma was induced by repeated sensitization and challenge with ovalbumin in the latter two groups. The dexamethasone intervention group received an intraperitonea injection of dexamethasone (2 mg/kg) before asthma challenge. Immunohistochemistry was used to measure the expression of Ang-1 and Tie-2 in the airway. Airway thickness was estimated by a computerized digital image analyzer.

RESULTSAirway thickness in the asthmatic group (33.9333+/-8.3791 micro m2/micro m) increased significantly compared with that in the control group (21.1333+/-2.7740 micro m2/micro m) (P<0.01). The dexamethasone intervention group also showed increased thickness of the airway (27.4000 +/- 4.6105 micro m2/micro m) compared with the control group (P<0.01), but the airway thickness in the dexamethasone intervention group was significantly reduced compared with that in the untreated asthmatic group (P<0.01). The expression of Ang-1 (103.9487+/-8.2914 vs 76.0320+/-3.7728; P<0.01) and Tie-2 (99.2307+/-8.1913 vs 75.3153+/-3.7321; P<0.01) in the airway increased significantly in the asthmatic group compared to controls. The expression of Ang-1 and Tie-2 in the airway of the dexamethasone intervention group (90.6180+/-5.2339 and 86.6633+/-3.7321, respectively) was statistically higher than that in the control group (P<0.01) but statistically lower than that in the untreated asthmatic group (P<0.01). Ang-1 and Tie-2 expression in the airway was positively correlated with the thickness of airway (r(Ang)-1=0.719r(Tie)-2=0.746P<0.01). There was also a positive correlation between Ang-1 and Tie-2 expression (r=0.742P<0.01).

CONCLUSIONSThe expression of Ang-1 and Tie-2 in the airway increased in asthmatic rats and was positively correlated with the thickness of the airway. Ang-1 and Tie-2 may participate in the process of airway remodeling in asthma. Dexamethasone can decrease the expression of Ang-1 and Tie-2 in the airway and relieve the changes of airway morphology.

Angiopoietin-1 ; analysis ; physiology ; Animals ; Asthma ; metabolism ; pathology ; Female ; Lung ; chemistry ; pathology ; Rats ; Rats, Sprague-Dawley ; Receptor, TIE-2 ; analysis ; physiology

OBJECTIVETo investigate the interaction between hydrogen sulfide (H2S)/cystathionine gamma-lyase (CSE) system and nitric oxide (NO)/nitric oxide synthase (NOS) system on cardiac protection in metabolic syndrome (MS) rats.

METHODSForty one male Sprague-Dawley rats were randomly divided into 6 groups: control group, MS group, H2S donor group, CSE inhibitor group, NOS inhibitor group, and NO donor group. The MS rat model was established by a high-fat diet of 16 weeks. Rats in control and MS groups were subjected to normal saline and the other four groups were respectively subjected to sodium hydrosulfide (NaHS, 56 μmol/kg), D,L-propargylglycine (PPG, 37.5 mg/kg), Nψ-nitro-L-arginine methyl ester (L-NAME, 18 mg/kg), L-Arginine (500 mg/kg) every day. Four weeks later, the obesity indices, blood sugar of oral glucose tolerance test in each time point (0,30,60, and 120 minutes) and blood lipids (cholesterol, triglyceride, high density lipoprotein, low density lipoprotein) were measured. The computer-based electrophysiological recorder system was used to measure the changes of the left ventricular systolic pressure (LVSP), the left ventricular end diastolic pressure (LVEDP), the maximal rate of pressure increase in the contraction phase (+dP/dtmax), and the maximal rate of pressure decrease in the diastole phase (-dP/dtmax). H2S and NO concentration in plasma and myocardium, as well as CSE, constitutive NOS (cNOS), and inducible NOS (iNOS) activities in myocardium were measured with colorimetric method. Reverse transcription-polymerase chain reaction was used to assess the gene expression of CSE and endothelial NOS (eNOS) mRNAs.

RESULTSCompared with control group, the obesity indices, blood sugar at each time point, and blood lipids significantly increased in MS group (P<0.05). H2S and NO concentration in plasma and myocardium, CSE and cNOS activities in myocardium, the expressions of CSE mRNA and eNOS mRNA, and the myocardial function significantly decreased in MS group (P<0.05). Compared with MS group, NO concentration in plasma and myocardium, cNOS and iNOS activities in myocardium, and the expression of eNOS mRNA significantly increased in CSE inhibitor group (P<0.05). However, activities of cNOS and iNOS in myocardium and the expression of eNOS mRNA were significantly decreased in H2S donor group (P<0.01), while the myocardial function significantly increased (P<0.05). H2S concentration in plasma and myocardium, and the expression of CSE mRNA significantly increased in NOS inhibitor group (P<0.05). However, in NO donor group, the CSE activity in myocardium and the expression of CSE mRNA significantly decreased (P<0.05). And the myocardial function was improved significantly (P<0.05).

CONCLUSIONSBoth the H2S/CSE and NO/NOS systems appear to have a mutual down-regulation effect on myocardium in MS rats. Meanwhile, exogenous H2S and NO supplement is cardioprotective in rat model of MS.

Animals ; Cystathionine gamma-Lyase ; metabolism ; physiology ; Disease Models, Animal ; Heart ; physiopathology ; Hydrogen Sulfide ; metabolism ; Male ; Metabolic Syndrome ; metabolism ; physiopathology ; Myocardium ; metabolism ; Nitric Oxide ; metabolism ; physiology ; Nitric Oxide Synthase ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the dynamic changes that occur in T cell subsets, particularly involving the surface expression of programmed death 1 (PD-1), in response to pegylated (Peg)-interferon (IFN) a-2a therapy in patients with chronic hepatitis C virus (HCV) infection.

METHODSTwenty-five patients with HCV genotype 1b chronic infection and 10 healthy controls were enrolled in the study. All the HCV patients received combination antiviral therapy of Peg-IFNa-2a (180 mug/week) plus ribavirin. At treatment weeks 0 (baseline), 4, 12, 24 and 48, the level of PD-1 protein expression on the surface of total peripheral CD8+ and CD4+ T cells was determined by flow cytometry and the level of PD-1 mRNA expression in peripheral blood mononuclear cells (PBMCs) was determined by reverse transcription-polymerase chain reaction. Independent student's t-test were used to compare mean values between the two groups, repeat measure variance analysis was used to compare mean values among multiple groups, and Pearson's correlation coefficient was used to assess correlation significance.

RESULTSOver the course of antiviral therapy, the proportions of CD4+ T cells and CD8+ T cells, as well as the CD4+/CD8+ ratio, increased (F = 81.23, 39.28, and 7.01 respectively; all P less than 0.01). In contrast, the PD-1 protein expression frequency on CD4+ T cells and CD8+ T cells significantly declined (F = 100.11 and 158.40 respectively; all P less than 0.01). The PD-1-mRNA expression level in PBMCs was: 1.40+/-0.26 at baseline, 1.30+/-0.27 at week-4, 1.14+/-0.18 at week-12, 1.06+/-0.26 at week-24, and 0.83+/-0.25 at week-48 (F = 20.09; P less than 0.01). A positive correlation existed between the PD-1 protein expression frequencies on CD4+ T cells and CD8+ T cells and the HCV RNA load detected at baseline (r = 0.82 and 0.75 respectively; all P less than 0.01).

CONCLUSIONThe ability of Peg-IFN-a-2a-based antiviral therapy to suppress HCV replication may involve reduction of PD-1 protein expression on the surface of CD8+ T cells and CD4+ T cells.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; CD4-CD8 Ratio ; Case-Control Studies ; Female ; Hepatitis C, Chronic ; drug therapy ; immunology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Programmed Cell Death 1 Receptor ; metabolism ; Recombinant Proteins ; therapeutic use ; Ribavirin ; therapeutic use ; T-Lymphocyte Subsets ; metabolism ; Young Adult

OBJECTIVEThe ventral part of the medial prefrontal cortex (mPFC) plays an important role in initiation and control of voluntary movement, mood and cognition. However, after the degeneration of the nigrostriatal pathway, the neuronal activity of the ventral mPFC and the role of serotonin(1A) (5-hydroxytryptamine, 5-HT(1A)) receptors in the firing of the neurons are still unknown. The present study is to investigate the change of neuronal activity in the ventral mPFC and the effect of systemic administration of the selective 5-HT(1A) receptor antagonist WAY-100635 on the activity of the neurons in normal and 6-hydroxydopamine (6-OHDA)-lesioned rats.

METHODSSingle unit responses were recorded extracellularly with glass microelectrodes from ventral mPFC neurons in normal rats and 6-OHDA unilaterally lesioned rats in vivo.

RESULTS6-OHDA lesion of the substantia nigra pars compacta (SNc) significantly increased the firing rate with no change in the firing pattern of neurons of the ventral mPFC in rats. Systemic administration of WAY-100635 (0.1 mg/kg, i.v.) did not change the mean firing rate and firing pattern of ventral mPFC neurons in normal rats. In contrast, WAY-100635 significantly decreased the mean firing rate of the neurons in rats with 6-OHDA lesion of the SNc.

CONCLUSIONThese data suggest that the degeneration of the nigrostriatal pathway results in an increase of neuronal activity of ventral mPFC and dysfunction of 5-HT(1A) receptor.

Action Potentials ; Animals ; Disease Models, Animal ; Male ; Neostriatum ; physiology ; Neural Pathways ; drug effects ; physiology ; physiopathology ; Neurons ; drug effects ; physiology ; Parkinson Disease ; physiopathology ; Piperazines ; pharmacology ; Prefrontal Cortex ; cytology ; drug effects ; physiology ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptor, Serotonin, 5-HT1A ; metabolism ; Serotonin 5-HT1 Receptor Antagonists ; Serotonin Antagonists ; pharmacology ; Substantia Nigra ; physiology

OBJECTIVETo investigate the pre- and post-chemotherapy expression levels of Th1 and Th2 type cytokines in peripheral blood CD4(+) T lymphocytes, the changes of Th1/Th2 ratio and their clinical significance in patients with gastric cancer.

METHODSThe levels of specific cytokines in 60 gastric cancer patients were detected by flow cytometry before and after chemotherapy with FOLFOX4.

RESULTSThe level of IFN-gamma from peripheral blood CD4(+) T lymphocytes after chemotherapy in the whole group of gastric cancer patients was 11.4% +/- 5.0%, significantly higher than that (9.5% +/- 3.4%) before chemotherapy (P < 0.05). The level of IL-10 after chemotherapy was 3.6% +/- 1.2%, significantly lower than that (4.2% +/- 1.8%) before chemotherapy (P < 0.05). The ratio of Th1/Th2 (IFN-gamma/IL-4) after chemotherapy was 3.4 +/- 1.0 versus 3.4 +/- 1.6 before chemotherapy, without significant difference (P > 0.05). Interestingly, the levels of IFN-gamma and TNF-alpha of peripheral blood CD4(+) T lymphocytes in 15 gastric cancer patients who achieved partial response (PR) after chemotherapy were 14.8% +/- 8.0% and 5.9% +/- 2.0%, respectively, both were higher than that (6.9% +/- 2.5% and 4.2% +/- 1.3%) before chemotherapy (both P < 0.05). Furthermore, the ratio of Th1/Th2 (IFN-gamma/IL-4) after chemotherapy was 4.0 +/- 1.5, significantly higher than 2.5 +/- 1.2 before chemotherapy (P < 0.01).

CONCLUSIONEffective chemotherapy may reduce the tumor burden and relieve the shift of Th1/Th2 ratio. Improvement in immune function may be a very important therapeutic measure due to poor response of chemotherapy in gastric cancer patients.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Fluorouracil ; therapeutic use ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-4 ; blood ; Leucovorin ; therapeutic use ; Male ; Middle Aged ; Organoplatinum Compounds ; therapeutic use ; Remission Induction ; Stomach Neoplasms ; blood ; drug therapy ; pathology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Tumor Burden ; drug effects ; Tumor Necrosis Factor-alpha ; blood