To obtain high-yield avilamycin-producing strains,low energy N~+ ion implantation technology and screening of streptomycin-re- sistant mutants are used in the study on breeding mutation.The results show that,“saddle”region,which range is from 3?10~(15) to 5?10~(15) ions/cm~2,has got better induced mutation action.It also means that the strain's resistant mutation and yield mutation closely correlate to each other,and the method of streptomycin resistant screening is feasible.We have isolated a high-yield strain SVT-45 which the productivi- ty is 195% higher than the original strain's in the rotation-flask experiments.These results showed that the ion implantation was an effective method for microbe mutagensis.

OBJECTIVETo investigate the effects of RCCS on cell seeding onto 3-D scaffold and cell-scaffold composite culture in vitro.

METHODSRabbit skin fibroblasts of passage 2 were seeded at 2 x 10(6) cell per cm(3) onto/into polyglycolic acid (PGA) foams by static seeding (dropping a cell suspension onto foams) or dynamic seeding (rotating PGA foams and a cell suspension in RCCS). Attachment of cells in foams was observed by cell-counting after trypsin digestion. The effects of culture condition were next studied by culturing cell-PGA complexes in RCCS versus static culture condition. Distribution and proliferation of cells in foams were investigated with MTT, stereomicroscope and scan electron microscope.

RESULTSNumbers of cells adhering to polymers increased gradually during an initial period of 24 hours. Eight, 12 and 24 hours after seeding, the rates of adhering cells were significantly higher in the dynamic seeding group than in the static seeding group (46.70% + 2.16% vs. 31.50% +/- 3.54%; 56.36% +/- 3.18% vs. 34.28% +/- 3.16%; 66.32% +/- 4.60% vs. 37.38% +/- 4.66%; P < 0.01). The dynamic culture method as compared to the static method resulted in new tissues with a higher cellularity and more uniform cell distribution during a 3 period of weeks.

CONCLUSIONSRCCS has advantages of promoting cell attachment, uniform migration and proliferation in polymer scaffolds and can be used for construction of 3-D cell-polymer tissues in vitro.

Animals ; Cell Adhesion ; Cell Culture Techniques ; methods ; Cell Division ; Cell Movement ; Fibroblasts ; cytology ; Polyglycolic Acid ; pharmacology ; Rabbits ; Time Factors

OBJECTIVETo investigate the ability of human dendritic cells (DCs) inducing specific T lymphocyte response and inhibit the expression of HBeAg and HBsAg in 2.2.15 cell culture supernatant.

METHODSDCs were prepared from peripheral blood mononuclear cells induced with granulocyte macrophage colony-stimulating factor(GM-CSF) and interleukin 4. DCs was impulsed with pure HBsAg before DCs maturation and cocultured with self-blood T lymphocyte, while DCs without pure HBsAg stimulated group, T lymphocyte group and only T lymphocyte group were prepared as control group. The culture supernatant of 2.2.15 cell with stimulated T lymphocytes was collected on day 1, day 3, day 5 and day 7, respectively. The expressed levels of HBeAg and HBsAg were detected by ELISA method.

RESULTSDCs after antigen stimulation had a strong ability to present antigen and induce immune activation, DCs after loading with antigen in normal control and chronic hepatitis patients group had stronger stimulative ability for T lymphocytes proliferation than that of DCs without loading with antigen and only T lymphocyte group(P less than 0.01). The stimulating ability of DCs had a positive correlation to the dosage of loaded antigen; CTLs produced as a result of DCs stimulation had a specific inhibitive effect on the expression of HBeAg in 2.2.15 cell supernatant,but not on the expression of HbsAg.

CONCLUSIONHuman dendritic cells stimulated with HBsAg in vitro can efficiently present antigen and stimulate the production of specific CTLs to HBV, which can efficiently inhibit the expression of HBeAg in 2.2.15 cell supernatant- DC vaccine may become an antiviral therapy strategy for chronic hepatitis B virus infected patients in future.

Adolescent ; Adult ; Antigen Presentation ; Cells, Cultured ; Child ; Dendritic Cells ; drug effects ; immunology ; Female ; Hepatitis B Surface Antigens ; pharmacology ; Hepatitis B, Chronic ; immunology ; Humans ; Lymphocyte Activation ; Male ; Middle Aged

Aged ; Biopsy, Needle ; methods ; Coal Mining ; Humans ; Lung ; pathology ; Lung Neoplasms ; complications ; diagnosis ; Middle Aged ; Pneumoconiosis ; complications

Animals ; Apoptosis ; Concanavalin A ; toxicity ; Liver ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; metabolism ; Nitric Oxide ; biosynthesis ; Tumor Necrosis Factor-alpha ; biosynthesis

OBJECTIVETo study the protective effects of salidroside on injury induced by hypoxia/hypoglycemia in cultured SH-SY5Y cell.

METHODApoptosis and intracellular free calcium concentration ([Ca2+]i) were measured by flow cytometry, morphological changes and neuronal necrosis were observed with fluorescence microscope, and the lactic dehydrogenate (LDH) release was measured by LDH kits.

RESULTHypoxia/hypoglycemia cultures for 2 hours induced neuronal apoptosis and necrosis. They were 18.59% (P < 0.01) and 4.94% (P < 0.01) respectively. There were morphological changes including chromatin condensation, nuclear fragmentation and formed apoptotic bodies after exposed to hypoxia/hypoglycemia for 2, 4, 6, 12 hours. After 2 hours of hypoxia/hypoglycemia, neuronal [Ca2+]i and the release of LDH were significantly increased. They were 8.46 nmol/L (P < 0.01) and 16.59% (P < 0.01) respectively. The effects were enhanced with the extending time of hypoxia/hypoglycemia. Salidroside might have significantly decreased the percentage of neuronal apoptosis and necrosis, reduced neuronal [Ca2+]i and the release of LDH. The effects of salidroside were strengthened with the increasing of Salidroside dosage.

CONCLUSIONSalidroside has effect of anti-neuronal apoptosis. This effect might be related to its function of decreasing intracellular free calcium concentration.

Apoptosis ; drug effects ; Calcium ; metabolism ; Cell Hypoxia ; drug effects ; Glucosides ; isolation & purification ; pharmacology ; Humans ; Hypoglycemia ; metabolism ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Phenols ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rhodiola ; chemistry

OBJECTIVETo identify a novel HLA-A allele in a Chinese Han individual.

METHODSOne mismatch was observed in HLA-A locus in HLA typing for CMDP donors using bi-allelic SBT kit. A confirmatory test for novel HLA allele was performed with mono-allelic SBT kit.

RESULTSThe DNA sequence was confirmed to be a novel HLA-A allele. There was 1 nucleotide differed from the closest matching HLA-A*11:01:01 at position 393(G→A), which resulting a change from GGG to GGA at codon 107, led to a silent mutation, conserving the amino acid Gly.

CONCLUSIONA novel HLA-A allele was confirmed and officially named HLA-A*11:01:37 (Genbank accession number, JN209962) by the WHO Nomenclature Committee for Factors of the HLA System in January 2012.

Alleles ; Base Sequence ; Blood Donors ; HLA-A11 Antigen ; genetics ; Humans ; Sequence Analysis, DNA

Objective To analyze and summarize the clinical manifestations,diagnosis and treatment of pure spinal epidural cavernous hemangiomas.Methods We presented a case series of 6 patients with pure spinal epidural cavernous hemangiomas in our hospital from April 2012 to November 2016 with previous articles discussing their clinical presentation,radiological characteristics,surgical technique,pathological features,and functional outcome.Results All patients were diagnosed as pure spinal epidural cavernous hemangiomas by pathology.The cavernous hemangiomas were totally cut and patients recovered well after operations.All patients gradually improved neurologically and achieved a good outcome with no recurrence during the follow-up for 13 to 68 months.Conclusion Pure spinal epidural cavernous hemangioma is rare.It is easily confused with other spinal diseases.MRI is an important imaging examination method.Pathology is the method of diagnosis.Surgical resection is the main method of treatment,and the prognosis is good.

Objective To study the screening of trace amount mutation of BRAF V 600E gene for avoiding the appearance of ineffective treatment in cancer patients .Methods The internal competitive amplification fragments were used to improve the inhibition of wild-type blocking (WTB) probe on wile-type BRAF V600E gene to increase the detection efficiency of BRAF V600E genotype of trace amount mutation occurrence .Re-sults When the template DNA concentration was 50 -200 ng/μL ,the constructed trace amount gene muta-tion real time fluorescence quantitative detection method could completely block the amplification of the wild-type BRAF V600E gene .The sensitivity of this assay reached as high as 0 .1％ ,which was in line with the sen-sitivity requirement for the gene trace amount mutation detection technique .In the colorectal biopsy tissues from 50 cases of suspected colorectal cancer ,8 cases (16 .0％ ) of BRAF V600E gene trace amount mutation were detected by using this constructed method ,which had higher detection rate .Conclusion The constructed gene trace amount mutation detection method can make the rapid ,simple and low cost quantitative analysis for BRAF V600E gene trace amount mutation in clinical samples .

A filtering algorithm is proposed to deal with the medical ultrasonic image series in video format, which uses the relativity in spatial domain, gray value domain and temporal domain simultaneously. For each frame image, the relativity in spatial domain and gray value domain is utilized to construct the adaptive neighborhood first. Then the spatial weighted and gray value weighted filtering is performed in this neighborhood. Finally, the temporal relativity between the adjacent frames is used to perform the temporal weighted filtering. All the weighted filtering in the three domains uses Gaussian kernel, thus the filtering sensitivity resulting from the threshold selection is reduced, and the stability of the algorithm is enhanced. As it can be seen from the experimental results, the three-domains filtering algorithm can suppress the noise effectively, and the edge details can be reserved well. So it is useful for the feature extraction, recognition and analysis.
Algorithms ; Image Interpretation, Computer-Assisted ; methods ; Ultrasonography ; methods