NP, P and L gene of Newcastle disease virus of goose origin were amplified and cloned into pGEM-T easy vector and then subcloned into pCI-neo expression vector respectively, the positive clones were identified by enzyme cutting, PCR and sequencing. GFP reporter gene was inserted into the downstream of recombinant expression plasmid of P gene, which of stop codon was deleted. The experiment of transfection of P and GFP recombinant plasmid on COS-1 cells and CEF showed that GFP gene expressed, and this demonstrated that P gene was also expressed. This research may be helpful for further study of reverse genetics and functional genome of NDV of goose origin.

miR-200c has been shown to regulate the epithelial-mesenchymal transition (EMT) by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software (miRanda) was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.

OBJECTIVETo study the effects of S-3307 spraying time and density on photosynthetic characteristic of Ligusticum chuanxiong.

METHODThe photosynthetic characteristic of L. chuanxiong under different S-3307 spraying time and density was studied by plot cultivation experiment.

RESULTThe content of chlorophyll a and chlorophyll b in leaf increased when the spraying density was 20, 40, 80 mg x L(-1), while the net photosynthetic rate was the maximum. When the spraying density was 160 mg x L(-1), the content of chlorophyll a and chlorophyll as well as net photosynthetic rate were not increased.

CONCLUSIONS-3307 spraying can raise the photosynthetic capacity of L. chuanxiong and promote the form of assimilation products.

Chlorophyll ; metabolism ; Ligusticum ; drug effects ; metabolism ; Photosynthesis ; drug effects ; Plant Growth Regulators ; pharmacology

BACKGROUNDTo clarify the difference and significance of T-lymphocyte subsets in differential diagnosis between severe acute respiratory syndrome SARS) and common atypical pneumonia.

METHODSTotally 100 patients hospitalized in Beijing Ditan Hospital since March to June 2003 with clinical diagnosis of SARS were involved in this study. These patients courses of disease were over 3 weeks. These patients were divided into two groups, SARS group and common atypical pneumonia group (non-SARS group). The counts of CD3+, CD4+ and CD8+ T-lymphocyte of two groups were systematically recorded and analyzed.

RESULTSSixty-five of the patients were confirmed to have common type of SARS, including 26 males and 39 females, 50 cases received methylprednisolone treatment. Thirty-five cases had common atypical pneumonia (non-SARS), 21 were males while 14 were females, 20 cases received methylprednisolone treatment. All the cases of two groups were cured in the end. The SARS patients T-lymphocyte counts decreased first and then increased. Before 15 days of disease course, mean CD3+, CD4+, CD8+ T-lymphocyte counts of SARS patients were decreased apparently (694+/-568/microl, 441+/-356/microl, 309+/-462/microl). After 15th day of disease course, the counts gradually returned to normal CD3+, CD4+, CD8+ T-lymphocyte counts of non-SARS patients were normal. Compared with patients of the same group who were not treated with glucocorticoids, T-lymphocyte counts of non-SARS patients treated with glucocorticoids had no obvious difference. But glucocorticoids had some effect on SARS patients recovery of cellular immune function, i.e., it delayed the recovery by about 6 days.

CONCLUSIONWith or without treatment with glucocorticoids,the lowered CD3+, CD4+, CD8+ T-lymphocyte counts in the early stage are of very important significance in differential diagnosis between severe acute respiratory syndrome and common atypical pneumonia.

Adult ; Diagnosis, Differential ; Female ; Humans ; Lymphocyte Count ; Male ; Middle Aged ; Pneumonia ; diagnosis ; immunology ; Severe Acute Respiratory Syndrome ; diagnosis ; immunology ; T-Lymphocyte Subsets ; immunology

In order to find the compound basis of Phlomis younghusbandii Mukerjee that related to pharmacodynamic action, various chromatographic techniques were used to separate and purify the constituents of this plant, and physicochemical and spectral data were used to identify the structures of obtained compounds. A new furanolabdane diterpene glycoside, named as phlomisoside F, was isolated and identified, which was 15,16-epoxy-8(9),13(16), 14-labdatrien-7-ketone-19-oic acid-beta-D-glucopyranosyl ester.
Diterpenes ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Phlomis ; chemistry ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry

The cardiovascular circulation feedback control treatment instrument (CFCTI) is an automatic feedback control treatment system, which has the function of monitoring, alarming, trouble self-diagnosis and testing on the line in the closed loop. The instrument is designed based on the successful clinical experiences and the data are inputted into the computer in real-time through a pressure sensor and A/D card. User interface window is set up for the doctor's choosing different medicine. The orders are outputted to control the dose of medicine through the transfusion system. The response to medicine is updated continually. CFCTI can avoid the man-made errors and the long interval of sampling. Its reliability and accuracy in rescuing the critical patients are much higher than the traditional methods.
Automation ; instrumentation ; Cardiovascular System ; Feedback ; Medication Systems

Catheter Ablation ; Child ; Heart Septal Defects, Atrial ; surgery ; Humans ; Imaging, Three-Dimensional ; Male ; Tachycardia, Supraventricular ; surgery ; Tomography, X-Ray Computed

To combine the fibrinolytic with anticoagulant activities for therapy of thrombotic deseases, a fusion protein made of tissue-type plasminogen activator (t-PA) and hirudin was constructed and expressed in chia pastoris. To improve thrombolytic properties of t-PA and reduce bleeding side effect of hirudin, FXa-recognition sequence was introduced between t-PA and hirudin molecules.The anticoagulant activity of hirudin can be target-released through cleavage of FXa at thrombus site. t-PA gene and hirudin gene with FXa-recognition sequence at its 5'-terminal were obtained by RT-PCR and PCR respectively. The fusion protein gene was cloned into plasmid pIC9K and electroporated into the genome of Pichia pastoris GS115. The expression of fusion protein was induced by methanol in shaking flask and secreted into the culture medium. Two forms of the fusion protein, single-chain and double-chain linked by a disulfide bond (due to the cleveage of t-PA at Arg275-Ile276), were obtained. The intact fusion protein retained the fibrinolytic activity but lacked any anticoagulant activity. After cleavage by FXa, the fusion protein liberated intact free hirudin to exert its anticoagulant activity. So, the fusion protein is a bifunctional molecule having good prospect to develop into a new targeted therapeutic agent with reduced bleeding side effect for thrombotic diseases.
Animals ; Cloning, Molecular ; Electroporation ; Hirudins ; biosynthesis ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Plasminogen Activator ; biosynthesis ; genetics

Oral epidemic diseases of exposure personnel in long-term low-dose radiation yet have rarely been studied. Referred to WHO oral health survey method and symptom grading standard, data of 341 exposure persons in long-term low-dose radiation including α particle, β particle, and γ rays, etc., were collected from one camp in China in 2011 with cluster sampling and analyzed? with Foxpro 6.0 and SPSS 16.0 software. The exposure persons worked in low-dose radiation for a long time aged between 23 and 56, whose average age were 27.1 years old.In addition, their lengths of service were from 2 to 34 years (average 7.9 years) and average exposure time was 8 hours a day each year for more than three months. Average annual radiation dose equivalent was from 1.8 to 16.5 mSv (average 7.3 mSv). Total radiation dose equivalent was from 3.8 to 425.0 mSv (average 97.3 mSv).
Adult ; Health Personnel ; Humans ; Middle Aged ; Occupational Exposure ; adverse effects ; Radiation Dosage ; Radiation Injuries ; epidemiology ; Stomatitis ; epidemiology

OBJECTIVETo study the role of MEKK2 on the production of IL-2 in Jurkat cells stimulated by PHA/anti-CD28 antibody.

METHODSThe MEKK2 and JNK kinase activities were measured in both dominant negative MEKK2 Jurkat (dnMEKK2 Jurkat) cells and parental Jurkat cells. The AP(1) and IL-2 promotor activities were measured by luciferase activity assay. The IL-2 mRNA and protein were detected by RT-PCR and Western blot.

RESULTSAfter stimulation by PHA/anti-CD28, JNK was activated in parental Jurkat cells but not in dnMEKK2 Jurkat cells. The luciferase report gene activities of AP1 and IL-2 promotors were increased by 4- and 5-folds in parental cells whereas only by 1 fold in dnMEKK2 Jurkat cells. The level of IL-2 mRNA and IL-2 protein were increased in parental Jurkat cells but not in dnMEKK2 Jurkat cells.

CONCLUSIONMEKK2 plays an important role on the production of IL-2 in Jurkat cell stimulated with PHA/anti-CD28 antibody. It is a potential drug target for the treatment of GVHD and autoimmune disease.

Humans ; Interleukin-2 ; biosynthesis ; genetics ; Jurkat Cells ; MAP Kinase Kinase 4 ; metabolism ; MAP Kinase Kinase Kinase 2 ; metabolism ; physiology