Objective:To obtain the high-efficiency expression of the biological active rabies virus nucleoprotein in the prokaryotic expression system.Methods:This experiment uses codon optimization technology to re-encode the nucleoprotein gene of rabies virus CTN-1 strain, artificially synthesize the full-length gene and clone it into pET-43.1a prokaryotic expression vector, induced expression in BL21 (DE3) strain of Escherichia coli( E. coli), and used Western blot to detect its reactogenicity. Results:The results showed that after induction, SDS-PAGE electrophoresis analysis showed that an obvious expression band appeared at a molecular weight of 50×10 3, which was consistent with the expected protein band size. Among them, the E. coli concentration A600 is about 0.5, and the expression yield is the highest (about 32.3%) when induced at 37℃ for 5 h. Nucleoprotein expression product is mainly inclusion body when it is expressed in large quantities. After purification by Ni 2+ chelating chromatography, the purity of the target protein can reach over 95%. The purified product was identified by Western blot and positively reacted with the sera of mice immunized with rabies vaccine, indicating that the prokaryotic expression of the CTN-1 strain nucleoprotein has biological activity. Conclusions:This experiment successfully established a high-efficiency expression method for the nucleoprotein of the CTN-1 strain in the prokaryotic expression system, and obtained high-purity target protein, which provides a basis for further clinical diagnosis and preparation of new vaccines.

Adenine ; analogs & derivatives ; Fanconi Syndrome ; Humans ; Organophosphonates

Carcinoma ; genetics ; metabolism ; pathology ; surgery ; China ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; metabolism ; pathology ; surgery ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-ets ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serine Endopeptidases ; genetics ; metabolism

Objective To investigate the effects of microRNA-155 (miR-155) inhibitor on JAK/STAT1 (Janus kinase/signal transducer and activator transcription 1) signaling pathways in the injured lung tissue induced by lipopolysaccharide (LPS).Methods One hundred and twenty BALB/c mice were randomly divided into control group (n =40),LPS group (n =40),and inhibitor + LPS group (n =40).LPS group and inhibitor + LPS group were made by injection of LPS 20 mg/kg intra-peritonealy,whereas equivalent volume of normal saline was given instead in the control group.The 80 mg/kg of miR-155 inhibitor was injected into caudal vein 24 h before LPS injection in inhibitor + LPS group.Mice were sacrificed at 6 h,12 h,24 h,and 48h separately after LPS injection,and lung tissue were collected.The levels of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) of lung tissue were measured using the enzyme-linked immunosorbent assay (ELISA).Using histopathological examination,the injury of lung tissue was evaluated.The expressions of miR-155,STAT1 mRNA,SOCS1 mRNA in lung tissue were assayed by fluorescent quantitative reverse transcription polymerase chain reaction (qRT-PCR).Results The miR-155 expression induced by LPS increased at 6 h,12 h,24 h and decreased at 48 h.The miR-155 expressions in LPS group were (8.52 ± 1.12) at 6 h,(11.04 ±0.99) at 12 h,(15.84 ±0.80) at 24 h and (4.03 ± 2.55) at 48 h.In the inhibitor + LPS group,the expressions of miR-155 were lower compared with LPS group,showing significant differences at 12 h (t =6.08,P ＜ 0.01),and at 24 h (t =23.64,P ＜ 0.01).STAT1 mRNA and SOCS1 mRNA both reached peak levels at 6 h after LPS injection.The levels of STAT1 mRNA in LPS group were higher than those in inhibitor + LPS group,showing significant differencesat6h (t=4.41,P＜0.01),12h(t=2.69,P＜0.05),and24h (t=3.62,P＜0.01).The levels of SOCS1 mRNA in inhibitor + LPS group were higher than those in LPS group,showing significant differences at 6 h (t =4.55,P ＜0.01),12 h (t =4.12,P ＜0.01),24 h (t =2.38,P ＜ 0.05).TNF-α reached its peak value at 6 hours and IL-10 reached its peak value at 48 hours.Both TNF-α and IL-10 were higher in LPS group than those in inhibitor + LPS group showing significant differences at 6 h,12 h,24 h (P ＜0.01).The pathologic examination indicated the lung injury in inhibitor + LPS group was milder than that in LPS group.Conclusion The miR-155 increased in lung tissue of endotoxemic mice.miR-155 inhibitor may suppress JAK/STAT1 signaling pathway and protect the lung tissue.

In order to investigate whether cytokine-induced killer (CIK) cells can induce apoptosis of bcr-abl(+) K562 cells, apoptosis of K562 cells and CEM cells induced by CIK cells, etoposide or camptothecin was detected with flow cytometry DNA assay. RT-PCR showed that K562 cells expressed the bcr-abl fusion gene, K 562 cells, K562 cells/etoposide or K562 cells/camptothecin groups showed no sub-G(1) peak. K562 cells/CIK cells group showed sub-G(1) peak (38.1%). CEM cells showed no sub-G(1) peak. CEM cells/camptothecin or CEM cells/etoposide groups showed sub-G(1) peak (23.5% or 32.3% respectively). CEM cells/CIK cells group showed sub-G(1) peak (45.4%). Etoposide or camptothecin did not induce apoptosis of K562 cells. CIK cells induce apoptosis of K562 cells. Bcr-abl fusion gene prevented apoptosis induced by etoposide or camptothecin, but did not prevent apoptosis induced by CIK cells. This property may support the observed adoptive immunologic effect of allogeneic bone marrow transplantation and donor lymphocyte transfusions of CML case relapsing after allogeneic bone marrow transplantation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; immunology ; Camptothecin ; pharmacology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Etoposide ; pharmacology ; Flow Cytometry ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; K562 Cells ; drug effects ; immunology ; metabolism ; Killer Cells, Lymphokine-Activated ; cytology ; immunology

Objective To develop a magnetic resonance imaging (MRI) system for pelagic definitive treatment platforms, large support ships and ocean vessels.Methods The system was designed based on normal electromagnetic MRI technology, which was composed of six parts of gradient system, magnet system, coil system, radiofrequency system, spectrum control system and imaging system.The magnet system was developed with C-shaped open magnet,five systems were designed such as a super-stability excitation system, a super-sensitivity signal detection system, a magnetic filed dissipation precision control system, a shipborne sensitive equipment vibration control system and a software system, and a set of technical detection standards was proposed.Results The developed MRI system realized mobile imaging,gained advantages in imaging velocity,resolution,magnetic field dissipation control,ship bump resistance and environmental suitability,and thus could be used in marine mobile conditions. Conclusion The developed MRI system meets the requirements of pelagic medical treatment, and contributes to improving pelagic medical support ability of Chinese navy.

Adolescent ; Adult ; Endoscopy ; Ethmoid Sinus ; surgery ; Eyelids ; surgery ; Female ; Humans ; Male ; Middle Aged ; Orbital Fractures ; surgery ; Otorhinolaryngologic Surgical Procedures ; methods ; Young Adult

OBJECTIVETo review the clinical features of the thoracolumbar fracture with ankylosing spondylitis (AS) in order to avoid delayed or missed diagnosis.

METHODSFive patients of thoracolumbar fracture with AS treated from April 2005 to June 2007 in our department were studied retrospectively, male 4 cases, female 1 case, the age from 26- to 72-years-old with an average of 44.8 years. Analysis including: case history, number of the ankylosed vertebras, characteristic of fracture, active state rheumatism.

RESULTSThe patients had the history of AS for average 22.6 years. The mean number of the ankylosed vertebras was 18.2. Of the 5 cases, 1 case encountered traffic accident, 1case was sprained, and 3 cases without trauma were diagnosed as stress fracture. Two cases were trans-vertebra fracture: the fracture line was through T6, T7, or L1 vertebral body respectively; 3 cases were through the disc space: 2 cases were through L1,2 disc space, 1 case was through L2,3. No compression fracture and neurological injury were found. The acute inflammatory index such as ESR and CRP in 4 cases didn't correlate with the degree of pain. The non-steroidal anti-inflammatory drugs (NSAIDs) hadn't significant effectiveness in relieving pain. The patients were diagnosed as 'relapse' of AS in other hospital, and had been misdiagnosed for average 1.51 months.

CONCLUSION1) the fracture is prevalent at the middle or late period of AS when extensive ankylosis has been existed at the thoracolumbar region; 2) the fracture is common at the lower thoracal spine and the upper lumbar spine, and the majority is the stress fracture; 3) the fracture line may be through the vertebral body, but more often through the disc space; 4) it is like an exacerbation of AS and therefore to be missed diagnosis; 5) when the back pain exacerbated suddenly in the middle or late period of AS, the degree of pain not correlating with acute inflammatory index, and the NSAIDs ineffective, the thoracolumbar fracture should be considered.

Aged ; Female ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fractures ; diagnosis ; drug therapy ; etiology ; Spondylitis, Ankylosing ; complications ; Thoracic Vertebrae ; injuries

Background and purpose:In recent years, epigenetics research has become a new direction of cancer research. A large number of results have shown that the abnormal changes of epigenetic modifications have close connection with cancer. Genome-wide epigenetic modifications have become new markers for cancer. This study aimed to investigate the methylation of the promoter ofDBC1 gene in cervical cancer tissues of Uyghur women in Xinjiang, to explore the correlation between the gene methylation and the infection of HPV, and to evaluate whether it can be used as a tool with high sensitivity and specificity for cervical cancer screening.Methods:This study detected the infection of HPV16, 18 in 43 normal cervical tissues, 35 cervical intraepithelial neoplasia tissues and 54 cervical cancer tissues using the polymerase chain reaction (PCR) method. The methylation of the promoter ofDBC1 gene in above-mentioned tissues was detected by the methylation-specific PCR method. The expression ofDBC1 at mRNA level was measured by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) in 10 methylation-negative normal cervical tissues and 10 methylation-positive cervical cancer tissues.Results:In normal cervical tissues, CIN tissues and cervical cancer tissues, the infection ratios of HPV16 were 18.6%, 34.3% and 68.5%, respectively; the infection ratios of HPV18 were 2.3%, 8.6% and 16.7%, respectively; and the methylation ratios ofDBC1 gene were 23.3%, 40.0%, 87.0%, respectively. In 79 high-grade squamous intraepithelial lesions (CINⅡ and Ⅲ) and cervical cancer tissues, 50 of 79 were infected with HPV16/18, while 29 of 79 were negative. The methylation ratio ofDBC1 gene was 88.0% in HPV16/18 infection positive group while the methylation ratio was 55.2% in negative group (P<0.05). The expression ofDBC1 gene at mRNA level in 10 methy- lation-positive cervical cancer tissues was significantly lower than that in the 10 methylation-negative normal cervical tissues (P<0.05).Conclusion:The methylation ofDBC1 gene may become a molecular marker to detect cervical cancer of Uyghur women in Xinjiang.DBC1 gene methylation combined with HPV16/18 infection test can be used to aid diagnosis of cervical cancer.

Objective To develop a kit of time?resolved fluoroimmunoassay(TRFIA)for detection of Schistosoma japonicum protein SjP38,and evaluate its effectiveness. Methods The anti 9G7 SjP38 monoclonal antibody was used as the capture anti?body coated with 96?hole plate,and the Eu3+labeled 1A6 monoclonal antibody was used as the detection antibody to establish the TRFIA SjP38 kit. In addition,the accuracy,sensitivity,precision,stability and coincidence rate to pathogenic diagnosis of the kit were evaluated. Results This established kit possessed high accuracy,wide linear range from 2 to 1 250 ng/ml,high sensitivity with the minimum detectable concentration of 0.14 ng/ml,and good precision(the coefficient variation of the intra?and inter?assay were 3.6%to 4.6%and 5.1%to 6.7%,respectively). The stability tests showed that the reagents could be stable for six months at 4℃,7 d at 37℃. The positive and negative corresponding rates to the pathogen detection method were 95%and 100%respectively. Conclusion All the performance and detection indicators of the kit have reached the requirements of clinical test,but its clinical application still needs further validation.