To construct a TK-/gG- mutant of pseudorabies virus, the gG-detected transfer vector pUSKKBB and genomic DNA of pseudorabies virus TK-/gG-/LacZ+ were co-transfected into IBRS-2 cells. Transfection progeny were plated onto PK-15 cells and incubated for 2 days under methylcellulose. Then the overlay was removed and replaced by 1% low melting point agarose in DMEM supplemented with 150 microg/mL X-gal. After 2 days, white plaques were screened for and purified 4 times. By PCR amplification of gG-deleted gene and LacZ gene, a recombinant virus with TK-/gG- phenotype was confirmed. Sequence of the PCR product revealed that there were 1,176 bp detection in gG gene of the PRV TK-/gG- mutant. Amplifying the gG-deleted gene of different generations of the TK-/gG- mutant showed that the mutant was stable within PK-15 cells. TCID50 assay indicated that the recombinant virus grows well on PK-15 cells. The mice immunized with the TK-/gG- virus showed no sign of abnormality. As a control, all mice inoculated with PRV strain died from the infection. All mice that received TK-/gG- survived after a lethal PRV challenge. However none of the mice injected with phosphate-buffer saline (PBS) survived from the challenge. The above results demonstrated that the recombinant virus could be a candidate marker vaccine strain for eradicating pseudorabies in pig herds.
Animals ; Herpesvirus 1, Suid ; genetics ; pathogenicity ; Mice ; Mice, Inbred BALB C ; Mutation ; Pseudorabies Vaccines ; immunology ; Swine ; Thymidine Kinase ; genetics ; Vaccines, Synthetic ; Viral Envelope Proteins ; genetics ; immunology

A coxsackievirus B strain was successfully isolated by cells culture from cardiac muscle tissues of a dead Sichuan golden monkey with myocarditis from a zoo of Changchun in China. The isolate was consistent with CVB by morphology, physicochemistry test, animal regression test and RT-PCR. Analysis of VP1 partial gene sequence and detection of mice specific serum IgG showed that the strain isolated was a coxsackievirus B3. It was the first CVB case report in Sichuan golden monkey and the strain isolated was named CVB/SGM-05.
Animals ; Cercopithecus aethiops ; Enterovirus B, Human ; isolation & purification ; Haplorhini ; virology ; Heart ; virology ; Mice ; Reverse Transcriptase Polymerase Chain Reaction ; Vero Cells ; Viral Structural Proteins ; genetics

OBJECTIVETo describe the case fatality rate of SARS in Beijing.

METHODSData of SARS cases notified from Beijing Center for Disease Control and Prevention (BCDC) and supplemented by other channels were collected. The data were analyzed by rate calculation.

RESULTSThe case fatality rate of SARS in Beijing was 7.66%, and had an ascending trend while the age of cases was getting older, and a descending trend while the epidemic development. The case fatality rate in Beijing was lower than that in other main epidemic countries or regions.

CONCLUSIONSThe risk of death increases with the increment of age of SARS patients. Beijing is successful in controlling and treating SARS.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Occupations ; Severe Acute Respiratory Syndrome ; mortality

OBJECTIVETo summarize lessons learned from an outbreak of severe acute respiratory syndrome (SARS) in China during the spring of 2004.

METHODSData of SARS cases were officially reported by Beijing Municipal Center for Disease Control and Prevention (BCDC) and Anhui Provincial Center for Disease Control and Prevention (APCDC) and results of epidemiological investigations were collected and analyzed.

RESULTSThree generations of 11 cases of SARS were identified during the outbreak. Initial two cases were most likely to be infected in Diarrhea Virus Laboratory of National Institute of Virology, China Centers for Disease Control and Prevention and main mode of transmission was direct contact with SARS patients. Delay in detecting initial case resulted in spread of the illness at hospitals and communities with two generations of secondary cases.

CONCLUSIONSSARS outbreak in 2004 has yielded following lessons for public health globally. (1) Lab bio-safety programs should be made and should be strictly abided by. Studies in highly pathogenic viruses such as SARS coronavirus should be utmost cautious. (2) Management systems of occupational exposure to virus and disease surveillance need to be strengthened to take all risk factors into account so as to detect potential patients with infectious disease as early as possible.

China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Male ; Occupational Exposure ; prevention & control ; Occupational Health ; Retrospective Studies ; Severe Acute Respiratory Syndrome ; epidemiology ; prevention & control ; transmission

Actinobacillus pleuropneumoniae ; drug effects ; genetics ; Animals ; Bacterial Proteins ; genetics ; Blotting, Western ; Drug Resistance, Bacterial ; genetics ; Green Fluorescent Proteins ; genetics ; Hemolysin Proteins ; genetics ; Hemolysis ; Mice ; Mutation ; Polymerase Chain Reaction

OBJECTIVETo assess the feasibility and accuracy of CT coronary angiography (CTCA) combined with adenosine stress myocardial perfusion scintigraphy (MPS) for diagnosis of flow-limiting coronary stenosis.

METHODSA total of 105 patients with suspected or established coronary artery disease (CAD) underwent CTCA and MPS within 4 weeks before invasive coronary angiography. The accuracy of CTCA/MPS in the diagnosis of flow-limiting coronary stenosis was evaluated in comparison with the results of quantitative coronary angiography and MPS.

RESULTSThe sensitivity, specificity, positive predictive value and negative predictive value of CTCA/MPS as a combined approach for detection of flow-limiting coronary stenosis were all 100%. In 16% (9/55) of the patients, revascularization procedures were performed and no flow-limiting stenosis was found.

CONCLUSIONCombination of CTCA and MPS has an excellent accuracy for detecting flow-limiting coronary stenosis as compared with quantitative coronary angiography/MPI, and can be a useful gatekeeper for revascularization procedures.

Adenosine ; Aged ; Coronary Angiography ; methods ; Coronary Stenosis ; diagnosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Myocardial Perfusion Imaging ; methods ; Tomography, Emission-Computed, Single-Photon ; methods ; Tomography, X-Ray Computed

Objective To investigate the potential possibilities of specific microRNA in plasma as novel biomarkers for the early diagnosis in juvenile idiopathic arthritis (JIA).Methods This research was be segmented into 4 stages.1.Screened candidate microRNAs:5 candidate plasma microRNAs were detected by biochips of microRNAs,corresponding 5 JIA patients with onset of oligoarthritis,5 JIA patients with onset of polyarthritis and 3 age-matched and sex-matched healthy controls individual.2.The feasibility of the microRNAs as novel biomarkers was validated by Realtime quantitative polymerase chain reaction (RT-PCR) in plasma on 80 JIA patients (43 oligoarthritis,37 polyarthritis),29 juvenile ankylosing spondylitis(JAS) patients and 30 healthy control individuals.3.The change of microRNAs was observed by RT-PCR in plasma from another 9 JIA patients before and 3 months after anti-rheumatic drug treatment.4.The correlation between the levels of candidate plasma microRNAs and clinical parameters of JIA patients was analyzed.Results Plasma concentrations of miR-16,miR-146a and miR-223 in JIA patients,including oligoarthritis and polyarthritis,were significantly higher than those in healthy subjects (all P ＜ 0.05) and JAS patients (all P ＜ 0.001),and plasma concentration of miR-132 in JIA were significantly lower than those in healthy subjects and JAS patients (all P ＜ 0.001).Although the plasma concentration of miR-16 in polyarthritis JIA patients was considerably higher than that in oligoarthritis JIA patients (all P ＜ 0.01),the plasma concentrations of miR-146a,miR-223 and miR-132 were no difference between the 2 subtypes of JIA(all P ＞0.05).More importantly,it was found that the expression of miR-16 was considerably reduced in the post-treatment plasma samples when compared to the pre-treatment samples(P =0.061).In addition,the levels of miR-16 in JIA plasma inversely corrected with tender joint.Conclusions Plasma levels of miR-16 were well-discriminated between JIA patients and healthy subjects or JAS patients.It is suggested that plasma miR-16 might serve as a novel and potential biomarker for screening JIA.Furthermore,it might serve as a novel biomarker for joint inflammation but not specifically for differentiating the subtypes of JIA.

Chronic hypoxic lung diseases are major causes of disability and mortality worldwide, which are typically aggravated by hypoxic pulmonary hypertension.The pathogenesis of hypoxic pulmonary hypertension is complex, and its mechanism has not been fully elucidated.The previous studies have shown abnormally elevated levels of free Ca + in the cytoplasm of pulmonary artery smooth muscle cells to be the predominant drivers of pulmonary hypertension , causing continuous contraction and remodeling of the pulmonary vessels.This article briefly summarizes the mechanism of hypoxia-induced imbalance in calcium homeostasis in pulmonary artery smooth muscle cells, together with its related drug research, based on the existing literature.Hypoxia induces an imbalance in calcium homeostasis in pulmonary artery smooth muscle cells by regulating hypoxia-inducible factor-1, K+ , store-operated calcium channel, receptor-operated calcium channel, the Ca +-sensing myosin contractile mechanism by binding to calmodulin, leading to pulmonary vasoconstriction.Ca + can also activate PKC/ MAPKs and PI3K/Akt/mTOR pathways, leading to pulmonary vascular remodeling.

Pulmonary hypertension(PH)is a chronic,progressive,high-mortality disease characterized by a continuous increase in pulmonary vascular pressure. All types of PH have the same characteristics,i.e.,the excessive proliferation,anti-apoptosis and inflammation of pulmonary artery endothelial cells and smooth muscle cells,which leads to progressive thickening of pulmonary small vessels,resulting in pulmonary vascular remodeling and increased pulmonary vascular resistance,ultimately leading to right ventricular hypertrophy,heart failure,and death. The drugs used to treat PH mainly include L-type calcium channel blockers,phosphodiesterase 5 inhibitors,guanosine cyclase activators,endothelin receptor antagonists,and synthetic prostacyclin and its analogues. These drugs reduce pulmonary artery pressure by relaxing pulmonary blood vessels but do not cure the patient,and their prognosis remains poor. Therefore,the development of drugs that can effectively improve or even reverse pulmonary vascular remodeling is the key to treating PH. In recent years,studies on pulmonary vascular remodeling mainly included(1)the synthesis of new small-molecule compounds;(2)the transformation of mature drugs,such as the use of drug combinations and dosage form transformation,etc.;(3)the pharmacodynamic evaluation of traditional Chinese medicines and derived compounds based on the theory of "lung distension";(4)research into monomers of traditional Chinese medicine; and(5)research into new targets.

Pharmacokinetic parameters can be significantly altered for acute kidney injury (AKI), extracorporeal membrane oxygenation (ECMO) and continuous veno-venous hemofiltration therapy (CVVH). Here we reported a case of individualized vancomycin dosing for a patient diagnosed as severe acute pancreatitis treated with concurrent ECMO and CVVH. A 65 kg 32-year-old woman was admitted to hospital presented with severe acute pancreatitis (SAP), respiratory failure, metabotropic acidosis and hyperkalemia. She was admitted to intensive care unit (ICU) on hospital day 1 and was initiated on CVVH. She progressed to multiple organ dysfunction syndrome (MODS) and acute respiratory distress syndrome (ARDS) on ICU day 2, and veno-venous ECMO was instituted. Several catheters were inserted into the body to support ECMO, CVVH and pulse indicator continuous cardiac output (PiCCO), so vancomycin was prescribed empirically on ICU day 3 for prevention of catheter-related infection. Given the residual renal function and continuous hemofiltration intensity on day 3, vancomycin bolus of 1 000 mg was prescribed, followed by a maintenance dose of 500 mg every 8 hours. On ICU day 4, a vancomycin trough serum concentration of 14.1 mg/L was obtained before the fourth dose, which was within the target range of 10-20 mg/L. By ICU day 7, vancomycin dosage was elevated to 1.0 g every 12 hours because of aggravated infection and improved kidney function. On ICU day 14, a vancomycin trough serum concentration of 17 mg/L was obtained. Her white blood cell (WBC) and neutrophil percentage (Neut%) dropped to the normal level by ICU day 19. This vancomycin regimen was successful in providing a target attainment of trough serum concentration ranging from 10-20 mg/L quickly and in controlling infection-related symptoms and signs properly. With the help of this case report we want to call attention to the clinically significant alteration in vancomycin pharmacokinetics among critically ill patients. Individualized vancomycin dosing regimens and therapeutic drug monitoring are necessary for critically ill patients receiving CVVH and ECMO to ensure that the target serum vancomycin levels are reached to adequately treat the infection and avoid nephrotoxicity.
Adult ; Anti-Bacterial Agents/administration & dosage* ; Critical Illness ; Extracorporeal Membrane Oxygenation ; Female ; Hemofiltration ; Humans ; Pancreatitis/drug therapy* ; Vancomycin/administration & dosage*