Hepatitis B is a serious infectious disease worldwide, and hepatitis B virus (HBV) is the direct cause of this disease. In recent years, as an essential part of its evolutionary process, HBV mutation has been extensively studied domestically and globally. However, the study on the conserved sequences in HBV sequences is still in its infancy. In this study, we applied multiple EM for motif elicitation (MEME) algorithm to discover HBV motif and proposed a new metric, conservative index (CI), to carry out phylogenetic analysis based on HBV sequences. Then, the constructed phylogenetic tree was subjected to reliability assessment. The results demonstrated that the new metric CI combined with the MEME algorithm can effectively help to discover motifs in HBV sequences and construct a phylogenetic tree based on them and to analyze the evolutionary relationship between HBV sequences; in addition, the possible ancestral sequences of samples may be obtained by conservative analysis. The proposed method is valuable for the exploratory study on large HBV sequence data sets.
Computational Biology ; Conserved Sequence ; Evolution, Molecular ; Hepatitis B virus ; genetics ; Humans ; Nucleotide Motifs ; Phylogeny ; Reproducibility of Results

Feeding Methods ; Humans ; Infant ; Male ; Mastication ; Syphilis ; etiology

OBJECTIVETo discuss the results and significance of the detection of the CFTR gene mutation in azoospermia patients with congenital unilateral absence of the vas deferens (CUAVD).

METHODSWe collected peripheral blood samples from 6 azoospermia patients with CUAVD for detection of the CFTR gene mutations and single nucleotide polymorphisms. We analyzed the genome sequences of the CFTR gene in comparison with the website of the UCSC Genome Browser on Human Dec. 2013 Assembly.

RESULTSMissense mutation of c. 592G > C in exon 6 was found in 1 of the 6 azoospermia patients with CUAVD and splicing mutation of c. 1210-12T[5] was observed in the noncoding region before exon 10 in 2 of the patients, both with the V470 haplotype in exon 11.

CONCLUSIONMutations of the CFTR gene can be detected in azoospermia patients with CUAVD and the detection of the CFTR gene mutation is necessary for these patients.

Azoospermia ; genetics ; Cystic Fibrosis Transmembrane Conductance Regulator ; genetics ; Exons ; Humans ; Male ; Male Urogenital Diseases ; genetics ; Mutation, Missense ; genetics ; Vas Deferens ; abnormalities

Hepatitis B is one of the most serious global threats to human health. Phylogenetic analysis of hepatitis B virus (HBV) can reveal the evolutionary relationship between HBV sequences and thus provide a basis for the prediction and treatment of hepatitis B and other aspects. In this study, we performed sequence analyses on the HBV sequences of five clinical HBV samples and the HBV sequences retrieved from the GenBank, EMBL, and DDBJ to construct a phylogenetic tree and analyze sequence structures. The experimental results revealed that the C gene of one cloned sequence had a recombinant structure of HBV B/ C subtype. Moreover, the phylogenetic results proved the existence of a newly found subtype HBV/B6 in Xishuangbanna of Yunnan Province, China. The experimental conclusion represents certain value for phylogenetic studies of HBV in Yunnan ethnic minority groups.
DNA, Recombinant ; genetics ; Genes, Viral ; genetics ; Genotyping Techniques ; Hepatitis B virus ; classification ; genetics ; Humans ; Phylogeny

OBJECTIVETo investigate the protective effects of daidzein (DD) on ventricular remodeling in rats with myocardial hypertrophy induced by pressure overload and its mechanism.

METHODMyocardial hypertrophy model of rats induced by pressure overload was prepared by constricting abdominal aorta. The operated rats were randomly divided into sham operated control group, aorta-constricted model group and three DD groups (30, 60, 120 mg kg(-1)). Four weeks later, the heart-weight (HW), left ventricular weight (LVW), the ratio of HW/BW and LVW/BW (LVI) and the cardio-myocyte diameters (MD) after dyeing by HE colar were measured. The hydroxyroline, nitric oxide (NO) and the activity of nitric oxide synthetase (NOS) and Na+ -K+ -ATPase, Ca2+ -ATPase in left ventricle were quantified with spectrophotometry and the angiotension II (Ang II) in left ventricle and serum was messured with radioimmunoassay.

RESULTAfter treatment of the left ventricular with DD, vs aorta-contricted model group, NO content, cNOS and Na+ -K+ -ATPase, Ca2+ -ATPase activity were significantly increased, the content of AngII in left ventricle and serum and iNOS activity and the ratio of HW/BW, LVI, MD were significantly reduced.

CONCLUSIONDD has protective effects on ventricular remodeling in rats with myocardial hypertrophy induced by pressure overload and its mechanism may be related to raising NO content and reducing the level of Ang II.

Angiotensin II ; blood ; metabolism ; Animals ; Hypertrophy, Left Ventricular ; metabolism ; pathology ; Isoflavones ; pharmacology ; Male ; Myocardium ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Phytoestrogens ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Ventricular Remodeling ; drug effects

BackgroundCorneal neovascularization (CNV) is a common eye disease.The researches on the pathogenesis and treatment of CNV are focus in Ophthalmology.ObjectiveThis study was to investigate the effects of culture supernatant from human amniotic epithelial cells (AECs) on CNV in vitro and its mechanism.MethodsHuman AECs were obtained from a placenta and cultured in serum-free medium for 48 hours,and the supernatant was collected.The levels of interleukin-1 receptor antagonist (IL-1 Ra) and pigment epithelium-derived factor (PEDF) in the human AECs culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA).Rabbit corneal epithelial cells (CECs) were obtained and cultured in different concentrations of human AECs culture supernatant for 48 hours,serum-free medium was used as the control group.The expression of vascular endothelial growth factor (VEGF) mRNA and basic fibroblast growth factor (bFGF) mRNA in rabbit CECs were measured by quantitative real-time PCR (QRT-PCR).Human umbilical cord vein endothelial cells (UVECs) were cultured in the three mediums above,and the proliferation of human UVECs (absorbance value,A value) was tested by the cell counting kit 8 ( CCK8 ).Migration assay was performed by the wound healing method for the human UVECs.The membrane ultra-structure of human UVECs was examined under the atomic force microscope (AFM).ResultsCultured and passaged human AECs showed a positive response for keratin.The expression of VEGF mRNA (1.00±0.22) and bFGF mRNA (1.00±0.36) in rabbit CECs was suppressed by the human AECs culture supernatant,with a significant reduction in comparison with the serum-free DMEM group (2.98±0.46,2.55±0.48 )(P=0.001,0.002).The A value was significantly lowered in the human AECs culture group for 72 hours compared with the serum-free DMEM group ( 1.941 ± 0.036 versus 2.144 ± 0.059 ) ( P =0.000 ),and the bFGF-induced migration rate of human UVECs was strongly inhibited by the culture supernatant of human AECs in comparison with serum-free DMEM.The plasma membrane of human UVECs cultured with the human AECs culture supernatant was full of bumps,and decreased intercellular connection and cellular pseudopodia were found on the AFM image.The concentration of IL-1Ra was (153.56±0.36)ng/L and that of PEDF was (70.41 ±0.68 )μ,g/L in the human AECs culture supernatant.Nothing was deteched in serum-free DMEM group.Conclusions Human AECs culture supernatant suppressed the expression of VEGF and bFGF in CECs as well as the migration and proliferation of vascular endothelial cells.The effect may be associated with IL-1Ra and PEDF secreted by human AECs.These results suggest that human AECs may be a potential therapy for the inhibition of CNV.

Objective To evaluate the role of expression of phosphodiesterase 4B (PDE4B) in the spinal cord in inflammatory responses in a rat model of neuropathic pain and the relationship with extracellular signal-regulated kinase (ERK).Methods Sixty healthy male Sprague-Dawley rats,weighing 180-220 g,in which intrathecal catheters were implanted at L5,6 interspace,were used.The location of catheters was confirmed 6 days later.The rats were randomly divided into 5 groups (n =12 each):sham operation group (group Sham),normal saline (NS) group,vehicle group (group Ⅴ),mismatch siRNA group (group siR-M),and PDE4B-siRNA group (group siR-B).Neuropathic pain was induced by ligation of L5 spinal nerve (SNL).In Sham group,the L5 spinal nerve was only exposed,but not ligated.Immediately after ligation and on 1,3,5,and 7 days after ligation,10 μl NS,10 μl NS,LipofectaminTM RNAiMAX,PDE4B-siRNA (2 μg/10 μl) encapsulated in mismatch siRNA and PDE4B-siRNA (2 μg/10 μl) encapsulated in LipofectaminTM RNAiMAX were injected intrathecally in Sham,NS,V,siR-M,and siR-B groups,respectively.The mechanical pain threshold was measured at 1 day before and 2,4,6 and 8 days after operation.After behavioral testing on 8th day after operation,the rats were sacrificed and the lumbar segment of the spinal cord was removed for determination of PDE4B protein,ERK and phosphor-ERK (p-ERK)expression,and TNF-α,IL-1β and IL-6 levels.Results Compared with Sham group,the mechanical pain threshold was significantly decreased at 2,4,6 and 8 days after operation in NS,V,siR-M and siR-B groups (P ＜0.05),and no significant change was found in the mechanical pain threshold at 2,4,6 and 8 days after operation (P ＞ 0.05) and the expression of p-ERK and PDE4B protein,and levels of TNF-α,IL-1β and IL-6 were increased at 2,4,6 and 8 days after operation in V and siR-M groups (P ＜ 0.05).Compared with NS group,the mechanical pain threshold was significantly increased,and the expression of p-ERK and PDE4B protein and levels of TNF-α,IL-1β and IL-6 were decreased at 2,4,6 and 8 days after operation (P ＜ 0.05),and no significant change was found in the parameters mentioned above in V and siR-M groups (P ＞ 0.05).Conclusion Up-regulation of the expression of PDE4B protein in the spinal cord is involved in the development of neuropathic pain in rats,which may be related to promoted phosphorylation of ERK in the spinal cord and enhanced inflammatory responses.

Objective To evaluate the role of phosphodiesterase 4B (PDE4B) in lipopolysaccharide (LPS)-induced release of inflammatory factors in rat microglias.Methods The pri mary cultured microglial cells were randomly divided into 5 groups (n =24 each) using a random number table:control group (group C),LPS group,vehicle group,mismatch small interfering RNA (siRNA) group and PDE4B-siRNA group.The cells were incubated for 48 h in C and LPS groups.The cells were transfected with lipofectaminTM RNAiMAX 1 μl for 48 h in vehicle group.In mismatch siRNA and PDE4B-siRNA groups,mismatch siRNA 2 μl and PDE4B-siRNA 2 μl were added to 100μl serum-free culture medium (final concentration of siRNA 20 nmol/L),respectively,lipofectaminTM RNAiMAX 1 μl was added simultaneously and the cells were then transfected for 48 h.The expression of PDE4B protein and mRNA was determined by Western blot and real-time PCR,respectively.The cells were cultured for 24 h in serum-free culture medium containing LPS 100 ng/ml in LPS,vehicle,mismatch siRNA and PDF4B-siRNA groups.Then the release of TNF-α and IL-1β was measured by ELISA and the expression of extracellular signalregulated protein kinase (ERK) and phosphorylated ERK (p-ERK) was detected by Western blot.Results Compared with C group,the expression of PDE4B protein and mRNA was significantly down-regulated in group PDE4BsiRNA (P ＜ 0.05),no significant changes were found in LPS,vehicle and mismatch siRNA groups (P ＞ 0.05),and the release of TNF-α and IU1β was increased and the expression of p-ERK was up-regulated in the other four groups (P ＜ 0.05).Compared with LPS group,the release of TNF-α and IL-1β was decreased and the expression of p-ERK was down-regulated in PDE4B-siRNA group,and no significant changes were found in vehicle and mismatch siRNA groups (P ＞ 0.05).There was no significant difference in ERK expression between the five groups (P ＞ 0.05).Conclusion PDF4B can promote LPS-induced release of inflammatory factors in rat microglias and activation of ERK is involved in the mechanism.

OBJECTIVETo explore the correlations between diagnostic information and therapeutic efficacy in rheumatoid arthritis (RA) with decision tree model analysis.

METHODSThree hundred and ninety seven patients came from 9 clinical centers were randomly divided into the Western medicine (WM) group (n=194) treated with non-steroidal anti-inflammatory drugs and slow-acting antirheumatic drug and the Chinese medicine (CM) group (n=203) with basic therapy and syndrome-differentiation dependant TCM treatment. TCM and WM diagnostic information were collected. The ACR 20 was used for efficacy evaluation and the information of patients before treatment was analyzed by SAS 8.2 statistical package. Through single-factor exploratory analysis, odds ratio of efficacy and variable was calculated taken P < 0.2 as the including criteria for data mining analysis with decision tree model. All data were classified into the training set (75%) and verifying set (25%) with efficacy as the variable for layering to make further verification of the data-mining analysis.

RESULTSTwenty variables were included in the CM group and 26 in the WM group in the data-mining model. In the former, 9 variables were positively correlated to the efficacy, including degree of arthralgia, tenderness and morning stiffness, number of swollen joint, and joint with tenderness, levels of IgM, rheumatoid factor (RF), C-reactive protein (CRP), and total assessment from doctor; and disease duration and degree of nocturnal polyuria were negatively correlated to that. While in the latter, 8 were positively correlated to the efficacy, including erythrocyte sedimentation rate (ESR), sour and weak waist and knees, white fur in tongue, joint ache and stiffness, swollen joint, and total assessment from doctor and patient, and red tongue with yellow fur and leucocyte count negatively correlated to it. Data mining with decision tree analysis revealed that different combinations of morning stiffness, slight red tongue, joint tenderness and nocturnal polyuria in the CM group, and those of white fur in tongue, CRP level, leucocyte count and morning stiffness in the WM group showed different efficacy, which were also verified in the randomly chosen verifying set.

CONCLUSIONTo analyze the correlations between diagnostic information and therapeutic efficacy with decision tree analysis is conformed to the theory of TCM in applying treatment according to syndrome differentiation individually, thus it would contribute to elevate the accuracy of therapy.

Adolescent ; Adult ; Aged ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Arthritis, Rheumatoid ; diagnosis ; drug therapy ; Decision Trees ; Diterpenes ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Epoxy Compounds ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phenanthrenes ; therapeutic use ; Phytotherapy ; Treatment Outcome

Vaccination is the primary strategy for the prevention and control of pandemic influenza. Because influenza virus is highly variable across strains, universal influenza vaccines need to be developed to address this problem. This review describes the research progress in conserved epitopes of influenza virus, the advances in the research and development of universal influenza vaccines based on the relatively conserved sequences of NP, M2e, HA2, and headless HA, the mechanisms of cross-protection, and the methods to improve cross-protection.
Animals ; Cross Reactions ; Humans ; Orthomyxoviridae ; immunology ; Species Specificity ; Viral Proteins ; immunology ; Viral Vaccines ; genetics ; immunology