Grass carp reovirus (GCRV), a disaster agent to aquatic animals, belongs to Genus Aquareovirus of family Reoviridea. Sequence analysis revealed GCRV genome segment 8 (s8) was 1296 bp nucleotides in length encoding an inner capsid protein VP6 of about 43kDa. To obtain in vitro non-fusion expression of a GCRV VP6 protein containing a molecular of fluorescence reporter, the recombinant baculovirus, which contained the GCRVs8 and eGFP (enhanced green fluorescence protein)genes, was constructed by using the Bac-to-Bac insect expression system. In this study, the whole GCRVs8 and eGFP genes, amplified by PCR, were constructed into a pFastBacDual vector under polyhedron (PH) and p10 promoters, respectively. The constructed dual recombinant plasmid (pFbDGCRVs8/eGFP) was transformed into DH10Bac cells to obtain recombinant Bacmid (AcGCRVs8/eGFP) by transposition. Finally, the recombinant bacluovirus (vAcGCRVs8/eGFP) was obtained from transfected Sf9 insect cells. The green fluorescence that was expressed by transfected Sf9 cells was initially observed 3 days post transfection, and gradually enhanced and extended around 5days culture in P1(Passage1) stock. The stable high level expression of recombinant protein was observed in P2 and subsequent passage budding virus (BV) stock. Additionally, PCR amplification from P1 and amplified P2 BV stock further confirmed the validity of the dual-recombinant baculovirus. Our results provide a foundation for expression and assembly of the GCRV structural protein in vitro.

AIM:To explore the differences between unilateral recess-resection (R & R) and bilateral lateral rectus recession (BLR-rec) in the treatment of basic intermittent exotropia.METHODS: A retrospective analysis of treatment of basic intermittent exotropia in 89 patients,in which 49 cases underwent unilateral recess-resection,40 cases underwent bilateral lateral rectus recession of external rectus retroperitoneal surgery January 2013 to January 2015 in our hospital.The stereopsis and strabismus were observed in 1d,1,6mo,1 and 2a after operation.RESULTS: There was no significant difference in the success rate and oblique degree between the two groups after 1d,1,6mo,1 and 2a (all P>0.05),but the success rate of the operation was reducing as time passed.After 2d of the operation,the drift of the R & R group was 12.10±5.74PD and the drift of the BLR-rec group was 7.78±4.21PD,the difference was statistically significant (P=0.021).The R & R group was more likely to cause lateral slanting than BLR-rec group.Two groups of patients with nearly stereopsis were both significantly improved,there was no significant difference between the two groups in the two groups (x2=4.530,P=0.210).CONCLUSION: The long-term stability of BLR-rec is superior to R & R.

3 years old than those in ≤3 years old group.The incidence rate of the adverse reaction after SPT was 1.6%(6/384),there were 4 temporary fieber,1 asthma and 1 anaphylactic shock.Conclusions The common allergens are inhalant allergens of dermatophagoides pteronyssinus and dermatophagoides farina in Chongming island.The SPT is more suitable for over 3 years old children with typical anaphylaxis of respiratory symptom and maybe have the potential danger.

Objective To assess left ventricular (LV) global systolic function in patients with essential hypertension with normal geometric LV by 3-dimensional ultrasound speckle tracking imaging(3D-STI). Methods Fifty patients with essential hypertension were enrolled in this study, 29 normal subjects matched with age and sex were selected as control groups. LV global longitudinal peak systolic strain (GLS), radial peak systolic strain (GRS), circumferential peak systolic strain (GCS), LV global 3-dimentional radial peak systolic strain (3DGRS) were measured in all subjects by 3D-STI from the apical full-volume image and compared between groups. LV ejection fraction (LVEF) was acquired from 3D-STI.Results Compared with controls, LV GLS, GRS, GCS and 3DGRS were significantly reduced in patients with hypertension ( P ＜ 0.05 for all). A Pearson correlate revealed that LV GCS, GLS and GRS corresponded with LVEF( r1 =0.930, P1 ＜0.001; r2 = 0.705, P2 ＜0.001; r3 =0.474, P3 =0.001,respectively) in patients with hypertension, and LV GCS, GLS correlated with LVEF( r1 = 0. 838, P1 ＜0. 001; r2 = 0. 697, P2 ＜ 0. 001, respectively) in normal subjects. Conclusions LV global 3D strain decreases in patients with hypertension in the early period,3D-STI could evaluate the early change of heart function in patients with hypertension. LV circumferential movement plays a major role in the LV 3D movement and impacts on LVEF.

AIM To establish an HPLC method for the simultaneous content determination of huperzine A and huperzine B from seventeen species Huperzioideae subfamily (including 2 ineditus new species).METHODS The analysises of tartaricacid extracts of huperzine A and huperzine B were performed on a room temperature column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of methanol-spiritus mindereri flowing at 1.0 mL/min in a isocratic elution manner,and the detection wavelength was set at 310 nm.The contents were determined by external standards.RESULTS Huperzine A and huperzine B showed good linear relationships within 0.002 0-0.30 μg,0.001 8-0.27 μg (r=0.999 9),whose average recoveries were 103.86％,101.3％ with the RSDs of 1.85％,1.30％,respectively.Huperzine A and huperzine B were found in seventeen species,and there were significant differences in contents from species to species.The content of huperzine A in Phlegmariurus hamiltonii was the highest,which reached up to 0.22％,with higher levels of it in Phlegmariurus cryptomerianus,Phlegmariurus petiolatus,and Phlegmariurus phlegmaria;The content of huperzine B was the highest in P.phlegmaria.CONCLUSION This accurate,stable and reproducible method can be used for the quality control of huperzine A and B from Huperzioideae subfamily.

Animals ; Aorta ; pathology ; Apolipoproteins E ; deficiency ; Atherosclerosis ; metabolism ; pathology ; Female ; Glycosides ; isolation & purification ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo understand if the Neuraminidase (N1) of Influenza A virus at the surface of yeast-displaying system, eukaryotic expression system and the infected cells could be used for anti-NA Abs screening, their activities and bindings to five candidate Abs were assayed.

METHODSThe surface NA expression was obtained by transfecting by recombinant NA constructors with specific tag-labels or live virus infection. The functional activity was measured by the fluorescent assay. Their bindings to the Abs were detected by flow cytometry.

RESULTSThe surface NAs presenting on the yeast-displaying system and eukaryotic expression system exhibited functional NA activities as the NA at the surface of virus-infected cells which showed affinities to Ab1, 4, and 5. The same bindings to Abl and 5 were found in the surface NA expressed by eukaryotic expression system while minor binding was observed in the yeast displayed-NA.

CONCLUSIONThe epitopes of yeast-displayed NA may be different from the NAs present at eukaryotic expression system and the infected cells which more likely suitable for the screening of anti-NA Abs.

Antibodies ; immunology ; Antigens, Surface ; genetics ; immunology ; Cell Line ; HEK293 Cells ; Humans ; Influenza A virus ; genetics ; immunology ; Neuraminidase ; genetics ; immunology ; Protein Binding ; Recombinant Proteins ; genetics ; immunology

Objective To investigate correlation between aspirin resistance(AR) and inflammatory factors. Methods One hundred and ten patients with coronary heart disease took aspirin 0.1 mg/d for 14 days.It was detected platelet aggregation function induced with adenosine disphosphate (ADP) and arachidonic acid (AA), and investigated correlation between AR and inflammatory factors. Interleukin-1? (IL-1?),interleukin-6 (IL-6) and high sensitive C-reaction protein (hs-CRP) levels. Results IL-6 level of patients with AR was significantly higher than that of aspirin sensitive (AS) patients. The other two index were not different between the two groups. Conclusion IL-6 levels could be used as predictor.

AIM:To investigate the expression of angiotensin-converting enzyme 2 (ACE2) in nephritic epithelium of primates. METHODS:The expression of ACE2 in Vero E6 cells was detected by the techniques of RT-PCR and immunocytochemistry (ICC) techniques. The distribution of ACE2 protein in kidney tissues of two Rhesus monkeys and two normal human cases was observed by immunohistochemistry (IHC) techniques. RESULTS:Vero E6 cells were found to express both ACE2 mRNA and protein. ACE2 protein was mainly located in epithelium of proximal tubules of Rhesus monkey and human kidney. CONCLUSION:The expression of ACE2 in epithelium of primate kidney may provide the condition for SARS-CoV entry,which may be one of the reasons for inducing renal damage in SARS patients.

Objective To explore the association of interleukin (IL)-1 genotypes with Alzheimer′s disease (AD). Methods Using polymerase chain reaction and restriction fragment length polymorphism, the IL-1A (-889) and IL-1B (+3953) genotypes in 84 cases of AD and 139 controls were detected and analyzed. Results The frequencies of IL-1A(- 889) C/C, C/T and T/T genotypes were 72.6% and 84.2%, 23.8% and 14.4%, 3.6% and 1.4% in AD cases and controls respectively. The genotypes frequencies of IL-1A (-889) C/C, C/T and T/T in AD cases were similar to that of controls (χ2=4.53, P＞0.05), but the frequencies of IL-1A (-889) T allele were significantly higher in AD cases than in controls (15.5% vs. 8.6%, χ2=4.93, P＜0.05). The frequencies of IL-1B (+3953) C/T genotypes and T allele were also significantly higher in AD cases than in controls (16.7% vs. 6.5%, 8.3% vs. 3.2%, χ2=5.88,5.56, both P＜0.05). Conclusions IL-1 genotypes are associated with AD. IL-1 genotypes may play an important role in the development of AD.