Objective To develop a new type of gene vaccine against plague. Methods caf1 gene of caf Operon Encoding F1 Antigen of Y. pestis was inserted into pHSS6-mTn-3xHA/lacZ library plasmids, the recombinant plasmids were linearized with Not I and transformed into yeast cell by lithium acetate (LiAc) method to induce homologous recombination. Positive recombinants were then selected with uracil-lack medium. Results These recombinants were confirmed by colony PCR to have the target gene fragments. Conclusion The present study provided a platform for constructing a novel expression system for expressing Y. pestis F1 Antigen via homologous recombination in Saccharomyces cerevisiae, which may contribute to the genetic prevention of plague through digestive-tract-route (DTR).

Based on the gene sequence of AIV matrix protein 2(M2) and two cytotoxic T-lymphocyte epitopes derived from nucleoprotein,the prokaryotic expression vector pET-3M2e-NP1-NP2 was constructed.The target gene was expressed in the solvable form in BL21(DE3) when induced with 1.0 mmol/L IPTG and Western blot analysis showed that the expression product had good immunogencity.Purified fusion protein was mixed with various amount of adjuvant,including Freund's,Vash oil and chitosan,and then immunized 20-day-old chicken by intramuscular injection and boosted 3weeks later.Blood samples were collected weekly following the primary vaccination.The anti-M2e antibody was detected with ELISA method with the synthesized peptide as coated antigen;the neutralizing ability of anti-serum was evaluated on MDCK cell line and chick embryo,the CD4+ and CD8+ T lymphocyte amounts in peripheral blood of immunized chicken was measured by flow cytometry.Results showed that the fusion protein can induce immunological reaction,and the antibody can bind with the viral M2 protein that expressed on the surface of MDCK cells.Flow cytometry result showed that CD4+ and CD8+ T lymphocyte in peripheral blood increased obviously following immunization(P

Textual researches of the origin of medical records indicate that the medical workers in the Qin and Han periods had definite purpose for collection of medical records and show important position of Records of the Historian in developing history of medical records. Analysis on concrete medical records indicates that medical records of acupuncture and moxibustion in Zhenji in writing also formed a certain style and reflected doctors' some trains of thought in clinical diagnosis and treatment of acupuncture and moxibustion.
Acupuncture Therapy ; history ; History, Medieval ; Humans ; Medical Records ; Medicine, Chinese Traditional ; history ; Moxibustion ; history

Adolescent ; Cerebellum ; diagnostic imaging ; pathology ; Cerebral Angiography ; Female ; Humans ; Intracranial Arteriovenous Malformations ; complications ; diagnostic imaging ; pathology ; Moyamoya Disease ; complications ; diagnostic imaging ; pathology

OBJECTIVETo construct a phosphatidylinositol 4-kinase beta (PI4K-beta) mutant with the 325th to 373rd amino acid codons deleted, and try to develop a simple method for constructing middle fragment deletion mutant.

METHODSIn line with the mechanism of gene splicing by overlap extension(SOE), an additional PCR was used to get the PI4K-beta mutant in which the 325th to 373rd amino acid codons were deleted. Then the mutated gene was cloned into pCMV-Tag4A mammalian expression vector.

RESULTSA mutant with the 325th to 373rd amino acid codons deleted was constructed successfully.

CONCLUSIONThe improved SOE is a very effective and reliable method to construct middle fragment deletion mutant. It is worthy to be popularized.

1-Phosphatidylinositol 4-Kinase ; genetics ; Base Sequence ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; genetics ; Polymerase Chain Reaction ; methods ; Protein Engineering ; methods ; Recombinant Proteins ; genetics ; Sequence Deletion

OBJECTIVETo describe factors causing the delay of diagnosis among non-resident tuberculosis (TB) patients and to study its implications from demographic, socio-economic aspects in order to provide information to policy makers for TB control programs in Shanghai.

METHODSA historical cohort study through questionnaire interview was conducted in 146 newly diagnosed TB patients. The questionnaire covered the general information of subjects and information on patients' health care seeking experiences from the first symptom to the TB diagnosis in health facilities. Index adopted to reflect the access to TB diagnosis would include the days due to delayed diagnosis consisting the days from both patient's and doctor's responsibilities.

RESULTSThe median days due to patients' delay was 19 (7-33.5) days, which was 3.8 times longer than 5 (2.5-10) days caused by doctor's. The median of total diagnostic delay was 31 (11-59) days. Female TB patients, patients with lower than annual 5000 Yuan income had an increased risk of a longer diagnostic delay with an OR of 3.226 and 11.958 relatively. Smear positive patients had a shorter delay (OR = 0.280, P < 0.05).

CONCLUSIONDelayed diagnosis was mostly caused by the patients, suggesting that TB control strategy for non-residents should aim at improving the access to TB diagnosis among patients with lower income and female non-residents.

China ; Data Collection ; Delayed Diagnosis ; Emigrants and Immigrants ; Female ; Humans ; Income ; Male ; Socioeconomic Factors ; Tuberculosis ; diagnosis ; economics

OBJECTIVETo observe the incidence, causes and deviation angle of axial offset in patients with fracture ununited treated by Ilizarov bone transport technology.

METHODSFrom January 2007 to December 2012, 10 patients with fracture ununited were treated by Ilizarov bone transport including 8 males and 2 females with an average age of (30.3 ± 10.6) years old ranging from 18 to 49 years old. The segment of bone defect involved upper tibial in 2 cases, medial tibia in 2 cases, lower tibial in 5 cases, upper femoral in 1 case. For Paley type of bone defect, 6 cases were type B1, 4 cases were B3. The incidence and deviation angle of axial offset after Ilizarov bone transport technology were observed and evaluated on bone result by Paley assessment.

RESULTSAll patients were followed up from 19 to 32 months with an average of (22.0 ± 5.6) months. Three cases were natural healed at fracture ends, the other 7 cases were healed after bone graft. The time of external fixator was 16 to 28 months. At the last follow-up, there were 3 cases occurred coronal angulation of angle 5° to 11° with an average of (8.7 ± 3.2). Sagittal angulation was in 4 cases, angle 6° to 9° with an average of (8.5 ± 2.1)°. There were 4 cases occurred axial offset. In the last follow-up, according to Paley evaluation criteria, osseous results were excellent in 7 cases, good in 3 cases; functional results were excellent in 6 cases, good in 4 cases.

CONCLUSIONAxial deviation after the Ilizarov bone transport treatment is relatively common, which will result in delayed healing of bone and poor limb alignment. In order to improve the bone healing, corresponding measurements should be taken to avoid or reduce the incidence of axial deviation during and after the operation.

Adolescent ; Adult ; Female ; Fracture Healing ; Fractures, Ununited ; surgery ; Humans ; Ilizarov Technique ; adverse effects ; Male ; Middle Aged

Objective To analyze the clinical characteristics and follow-up data of catheter ablation of recurrent atrial tachycardias (ATs) after Mini-Maze surgery,and to explore prognostic factors for recurrence.Methods 59 patients in Guangdong General Hospital with ATs post Mini-Maze and concomitant open-heart surgery from April.2010 to June.2015 were included.According to high density precise mapping,activation mapping,voltage mapping and entrainment mapping,they underwent electrophysiological study and ablation which was guided by three-dimensional mapping system.All patients were followed up regularly.We explored the prognostic factors for recurrence by the Cox regression analysis.Results There were 88 types of ATs being mappedwith mean (1.49 ± 0.75) types of ATs identified per case.Most ATs were macro-reentry ATs(67/88,76.1％)and focal ATs (20/88,22.7％),respectively.56 patients (94.9％) achieved immediate ablation success.In a mean follow-up of (30.8 ± 17.7) months,recurrences were observed in 12 patients after the first time catheter ablation.Recurrent time was 3.5 (1.3,12.0) months and the overall ablation success rate was 74.6％ (44/59).6 patients received second ablation and the achievement of freedom from arrhythmias reached 79.7％ (47/59).Multivariate analysis showed that the LA diameter was the independent predictor for recurrence (HR 1.108,95％ CI 1.002 to 1.226,P =0.045).Conclusion Catheter ablation of ATs post Mini-Maze with concomitant surgery is save and feasible.LA diameter is the independent predictor for recurrence.

Objective To establish the molecular characteristics of Mycobacterium tuberculosis and on factors influencing the recent transmission of drug resistant isolates in Shandong. Methods Mycobacterium tuberculosis isolated from active pulmonary tuberculosis patients of 13 counties were genotyped by mycobacterial interspersed repetitive units (MIRU) methods. Results 12 loci of MIRU were detected in 558 isolates and a total of 143 MIRU patterns were confirmed. 66 isolates had distinct patterns,and 481 (86.2%) strains were in clusters. Shandong cluster included 177 strains with 74.6% of the isolates belonged to Beijing family. The recent transmission index of multi-drug resistance strains was in lower level,comparing to the susceptible strains. Conclusion Our results showed that the Shandong cluster isolates had capacities of facilitating person-to-person transmission and high level of drug resistance.

OBJECTIVE: To determine the expressions of Notch1, Jagged1 and vascular endothelial growth factor (VEGF) in human non-small cell lung cancer (NSCLC) and to explore its clinical pathological significance. METHODS: Immunohistochemical SP method was used to detect the expressions of Notch1, Jagged1 and VEGF in 65 patients with NSCLC and 15 normal epithelial tissues of the lung, and the relationship between them and clinic-pathological parameters were analyzed. RESULTS: The positive rates of Notch1, Jagged1 and VEGF in NSCLC were 81.5%, 83.1% and 93.8%, respectively, higher than those in normal epithelial tissues of the lung (P<0.05). The positive expression levels of Notch1 and VEGF were closely associated with the tumor stage and the lymph node metastasis (P<0.05). The positive expression levels of Jagged1 was positively correlated with the pathological type and lymph node metastasis. There was a positive correlation between Notch1, Jagged1 and VEGF. CONCLUSION Notch1, Jagged1 and VEGF protein may play an important role in the pathway of carcinogenesis and progression of NSCLC. The up-regulation of Notch1, Jagged1 and VEGF protein expression probably predict NSCLC carrying relatively strong permeation and metastasis.
Adult ; Aged ; Calcium-Binding Proteins ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Jagged-1 Protein ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; Neoplasm Staging ; Receptor, Notch1 ; metabolism ; Serrate-Jagged Proteins ; Vascular Endothelial Growth Factor A ; metabolism