To improve the expression of heterologous genes using baculovirus expression system, we constructed a novel shuttle vector based on the Bm-Bacmid. In the Bm-Bacmid, partial sequences of Chitinase and Cystein Protease were replaced with a tandem cassette of Cm and egfp through homologous recombination. Bombyx mori bidensovirus (BmBDV) ns1 under the control of polyhedrin promoter was inserted into the modified Bm-bacmid by transposition. For comparison, BmBDV ns1 under the control of polyhedrin promoter was also cloned in the wild type Bm-bacmid. The resulting Bm-bacmids were transfected into the cultured BmN cells to prepare recombinant virus to infect silkworms for expression of BmBDV ns1. Total proteins of hemocyte from infected silkworms were subjected to Western blotting and ELISA analysis. The yield of BmBDV NS1 1 with the modified vector was three times as much as that with the unmodified vector. The method to improve the yield of BmBDV NS1 in silkworms will facilitate the function and three-dimensional structure study of BmBDV NS1.
Animals ; Baculoviridae ; Bombyx ; virology ; Cell Line ; Chitinases ; Cysteine Proteases ; Genetic Vectors ; Insect Viruses ; Promoter Regions, Genetic ; Transfection

Objective To characterize the molecular characteristics of hemagglutinin (HA) and neuraminidase (NA) of influenza B viruses isolated in Jiangsu province,2011.Methods Thirteen strains of influenza B virus in different areas and epidemic period in Jiangsu province,2011 were selected for whole-genome sequencing,and analysis of molecule epidemic characteristics for HA and NA was carried out by bioinformatics method.Results Of the 13 randomly selected influeuza B strains,10 strains were assorted to Victoria lineage strains with NA genes from Yamagata lineage,and 3 strains belong to Yamagata lineage.Compared nucleotide and amino acid sequences of HA and NA genes with their vaccine strains respectiuely,196/197 glycosylation site appeared on HA1 gene in Yamagata/Victoria isolates virus.Conclusion Both B/Victoria and B/Yamagata lineage viruses co-circulated in Jiangsu province,and reassortant virus of Victoria lineage were predominant virus.

OBJECTIVE: To analyze the effects of Astragalous Injection on oxidative stress and micro-inflammatory status in patients undergoing maintenance hemodialysis (MHD). METHODS: Sixty MHD patients were included and randomized into treatment group and control group, with another 10 healthy volunteers as normal control. The patients in the treatment group were treated with Astragalous Injection and the patients in the control group were treated with normal saline for 12 weeks. A spectrophotometric method was used for the measurement of plasma concentrations of oxidative parameters including advanced glycation end products (AGEs), advanced oxidation protein product (AOPP), malondialdehyde (MDA) and vitamin E (Vit E). The content of C-reactive protein (CRP) was evaluated by enzyme-linked immunosorbent assay. RESULTS: Compared with the normal control group, the plasma levels of AGEs, AOPP, MDA and CRP were significantly increased, while plasma level of Vit E was significantly decreased in MHD patients ( P<0.01). After Astragalous Injection treatment, the plasma levels of AGEs, AOPP, MDA and CRP were decreased as compared with the control group ( P<0.01), while there was no significant difference in plasma Vit E level between the treatment group and control group. CONCLUSION: There exist oxidative stress and micro-inflammation in MHD patients. Astragalous Injection can ameliorate the accumulation of oxidative products and micro-inflammatory status, but it has no significant effect on plasma Vit E level.

The present investigation was undertaken in order to select the surface-sterilization technique most efficient for eliminating epiphytes, to document endophytes of healthy tissues from Glaycyrrhiza inflat Bat. in Xinjiang. Surface sterilization with 5% commercial solution of sodium hypochlorite for 5 minutes was reaffirmed as adequate for removing epiphytes on licorice roots. From the 151 segments incubated, 149 bacterial isolates and 2 fungal isolates were obtained. From all the isolates, Bacterial isolates were identified by VITEK-AMS. Part of Bacteria were identified in 13 different genus. Fungal species were characterized as Penicillium sp. and Fusarium sp.with microscope.

Currently,three predominant subtypes of influenza virus are prevalent in pig populations worldwide:H1N1,H3N2,and H1N2.European avian-like H1N1 viruses,which were initially detected in European pig populations in 1979,have been circulating in pigs in eastern China since 2007.In this study,six influenza A viruses were isolated from 60 swine lung samples collected from January to April 2011 in eastern China.Based on whole genome sequencing,molecular characteristics of two isolates were determined.Phylogenetic analysis showed the eight genes of the two isolates were closely related to those of the avian-like H1N1 viruses circulating in pig populations,especially similar to those found in China.Four potential glycosylation sites were observed at positions 13,26,198,277 in the HA1 proteins of the two isolates.Due to the presence of a stop codon at codon 12,the isolates contained truncated PB1-F2 proteins.In this study,the isolates contained 591Q,627E and 701N in the polymerase subunit PB2,which had been shown to be determinants of virulence and host adaptation.The isolates also had a D rather than E at position 92 of the NS1,a marker of mammalian adaptation.Both isolates contained the GPKV motif at the PDZ ligand domain of the 3' end of the NS1,a characteristic marker of the European avian-like swine viruses since about 1999,which is distinct from those of avian,human and classical swine viruses.The M2 proteins of the isolates have the mutation (S31N),a characteristic marker of the European avian-like swine viruses since about 1987,which may confer resistance to amantadine and rimantadine antivirals.Our findings further emphasize the importance of surveillance on the genetic diversity of influenza A viruses in pigs,and raise more concerns about the occurrence of cross-species transmission events.

ObjectiveTo explore the effects of epidermal proteins and lamellar bodies on skin barrier in glucocorticoid-dependent dermatitis.MethodsTotally,60 patients with glucocorticoid-dependent dermatitis and 40 normal human controls were eligible for this study.A noninvasive method using TewameterTM was applied to determine transepidermal water loss (TEWL) value in these subjects.Tissue specimens were obtained from the lesions of 13 patients with glucocorticoid-dependent dermatitis and normal skin of 10 human controls.Subsequently,haematoxylin and eosin(HE) staining was performed to observe the histopathological changes,immunohistochemistry to detect the protein expressions of K6,K10,K14,K15,loricrin,filaggrin,involucrin in epidermis,and electron microscopy(EM) to estimate the density of lamellar bodies in tissue specimens.ResultsCompared with the normal controls,the patients displayed an elevated TEWL value (P ＜ 0.05),which suggested an impaired epidermal barrier.Histopathology of lesions revealed nonspecific inflammatorychanges withmarkeddifferencesbetweendifferentclinicaltypesofglucocorticoid-dependentdermatitis.Immunohistochemistry revealed an attenuated expression of K10,K14,loricrin,filaggrin,involucrin and abnormal expression of K15 in lesional epidermis compared with the normal epidermis (all P ＜ 0.05),hinting a suppression of epidermal differentiation and proliferation as well as an impairment of cornified envelope structure.The number and density of lamellar bodies were also reduced in lesional epidermis compared with the control epidermis.ConclusionsCompared with normal skin,the structure of skin barrier is impaired in lesions of glucocorticoid-dependent dermatitis,to restore skin barrier is essential for the treatment of this entity.

The aim of this study was to investigate the clinical, pathological and biological features of acute megakaryoblastic leukemia in childhood. The morphology of cells was observed by means of bone marrow smear; the immunophenotype was detected by flow cytometry and immunohistochemistry assay. The results indicated that the fever, hemorrhage, hepatosplenomegaly and lymphadenopathy in this case were the primary presentations accompanying by leukocytosis, anemia and thrombocytopenia. An adequate marrow aspirate could not be obtained. At the time of diagnosis, the bone marrow had more than 30% megakaryoblasts in nucleated cells. Flow cytometric analysis revealed the dual expression of CD41 and CD61 by tumor cells in bone marrow. The histopathological examination of bone marrow demonstrated infiltration of large-sized CD42b(+) cells. According to all above mentioned results, this case was diagnosed as acute megakaryoblastic leukemia. In conclusion, childhood acute megakaryoblastic leukemia is a rare and easily misdiagnosed disease with poor prognosis. Flow cytometry analysis and immunohistochemistry assay of bone marrow can help in detecting this leukemia subtype and evaluating its prognosis.
Bone Marrow Cells ; immunology ; pathology ; Female ; Flow Cytometry ; Humans ; Infant ; Leukemia, Megakaryoblastic, Acute ; diagnosis ; immunology ; pathology ; Platelet Glycoprotein GPIb-IX Complex ; immunology

Objective To provide structure and function applied anatomical of nasal sub-units for plastic and reconstruction. Methods Twelve cases fresh nose from adult head specimen,to dissect the skin,muscle fascia system, cartilage and bone,to observe and digiital measure nerve, blood vessel and ligament between cartilage. Results The thinnest skin in the point of bone and cartilage junction,the thickest parts in the nasion and the supratip breakpoint.Nose contours include of the skin,cartilage,bones and vascular muscle fascia system;Nasal subunit can be divided into nasion area,nasal dorsum area, nasal tip area,nasal ala area and nasal columella area;Nasal valve was a important anatomic part of bremh.The nasal lateral osteotomy of maxilla can change 2 mm height and width of the nasal bone;Lower lateral cartilage and upper nasal cartilage connection can be separated 6~8 mm. Curvature changes in the crura intermedium of alar cartilage Can raise nasal tip 2 mm. Extent of septal cartilage was 15 mm×20 mm,thickness was 1 mm.The mainly blood supply come from facial artery and ophthalmic artery. Vein accompanying with the same name.Lymphatic flow in the area of superficial muscle fascia. Concision Nasal bone lateral osteotomy can raise up bridge of the nose.To lengthen and highten nasal tip based on the complete strip technique of lower lateral cartilage and upper lateral,and to change angel of medial crus and middle crus of lower lateral cartilage by suture rotation.Nasal skin is rich in artery and vein,so much as four networks.Because the syndrome of nasal contraction deformation occurred more and more recently we must to avoid the vascular network injury when operation, and the anatomical level should be give more noted.

Objective To study the biological characteristics of arsenic resistance cell model chronic arsenic exposure human bone marrow mesenchymal stem cells (CAsE-hFBMSCs) and discuss the consequence of chronic arsenite exposure to human mesenchymal stem cells (hFBMSCs). Methods hFBMSCs cultivated under general conditions,hFBMSC cell survival rate was detected in 48 hours with arsenite toxicity test under different doses arsenic [0(control),0.25,0.50,1.00,2.00,4.00,8.00,20.00,40.00,80.00,120.00 μmol/L]of the fist 2-generation(P2). According to the test results,1.00 μmol/L sodium arsenite was chosen to stimulate hFBMSCs for 14 weeks as experimental group,simultaneous 0 μmol/L sodium arsenite as the control group. And then,the phenotype was detected by fluorescence-activated cell sorting,and the cell cycle by flow cytometry. Finally,the cell malignant transformation was detected by soft-agar assay. Results Arsenite low than 10 μmol/L promoted cell proliferation,but inhibited cell proliferation when exceeding 10 μmol/L. Half of the lethal dose (LC_(50)) in experimental and control groups were (89.42±0.64),(52.48±0.71)μmol/L. The difference between two groups was statistically significant(t = 123.89,P < 0.05). The phenotype of CAsE-hFBMSCs was CD29,CD90,CD166 positive and CD34,CD45 negative. The phenotype of CAsE-hFBMSCs was the same as the control. Comparing to control group[(8.44±0.45)%,(9.14μ0.14)%,(82.42±0.60)%],G2/M phage[(17.72±5.47)%]and S phage [(25.34±3.36)%]cell increased,G0/G1 phage[(56.96±8.83)%]cell decreased in P2 CAsE-hFBMSCs. The cell cycle became nearly the same as the control group after adaption. CAsE-hFBMSCs did not show clone formation in soft agar clone formation assay. Conclusion Long last and low level exposure to arsenite does not influence the biologic features of hFBMSCs.

Child ; Follow-Up Studies ; Humans ; Neoplasm Recurrence, Local ; Neuroblastoma ; Recurrence ; Treatment Outcome