To explore the feasibility of stable expression of Hantavirus H8205 strain G1 segment and human IL-2 fusion gene in Vero cells, and to examine the immune protection effects on mice vaccinated with this recombinant eukaryotic expression vector containing Hantavirus G1 gene and IL-2 gene. With the help of lipofectamine, the Vero cells were transfected with pcDNA3.1/HisB-IL-2-G1 and the positive cells were selected by G418. IFAT and SDS-PAGE electrophoresis were used to determine the stable transfection and expression of recombinant protein. Each mouse was inoculated with plasmids intramuscularly (i.m.) three times, 2 boosts were given at 2-week intervals, serum anti-hantavirus antibodies were detected by ELISA and neutralizing antibodies (NAb) were detected by Plaque Reduction Neutralization Test. The fusion protein expressed in Vero cells was 78 kD, corresponding to the estimated molecular size. The neutralizing antibody titers of mice with pcDNA3.1/HisB-IL-2-G1 were 1:20-1:80. IL-2/G1 fusion gene could be transferred in Vero cells and stably express the fusion protein. Specific humeral immune responses in mice can be induced with the recombinant eukaryotic expression vector containing the fusion gene, which lays the foundation for further development of therapeutic HTNV vaccine.

AIM: To investigate the effect of antisense oligonucleotides(ASON) of c-sis on cellular cycle and proliferation of pulmonary artery vascular smooth muscle cells(VSMC).METHODS: Tissue mass culture was done to get VSMC of pulmonary artery. Different concentrations of antisense oligonucleotides of c-sis were added into the cultures to observe the VSMC proliferation curve using MTT test. The changes of VSMC cellular cycle were also observed by flow cytometry. RESULTS: ASON with mid-to high concentrations restrained the proliferation of VSMC apparently with the peak of cell growth being attenuated or eliminated. Affected by mid-concentration ASON, PDGF-BB showed significant accelerating effect on the proliferation of VSMC. The ratio of G 0/G 1 in cellular cycle was increased significantly in VSMC culture with ASON in comparison with control. The G 0/G 1 ratio also showed significant differences among different concentration of ASON groups( P

Adult ; Angiomatosis ; immunology ; pathology ; surgery ; Antigens, CD ; metabolism ; Diagnosis, Differential ; Female ; Hemangioma ; immunology ; pathology ; Humans ; Hyperplasia ; Spleen ; immunology ; pathology ; Splenectomy ; Splenic Diseases ; immunology ; pathology ; surgery ; Splenic Neoplasms ; immunology ; pathology

China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Disease Outbreaks ; Humans ; Norovirus

Objective To investigate the role of negative-regulatory factors of toll-like receptors (TLRs)signal pathways in immunological pathogenesis of Kawasaki disease(KD).Methods Thirty-two chil- dren with Kawasaki disease and 16 age-matched healthy children were studied.Reverse-transcription PCR (RT-PCR)and real-time PCR were used to evaluate the mRNA expression levels of toll-like receptor 4(TLR4), MD-2,MyD88,IRAK-4,TRAF6,T1/ST2,IRAK-M,Triad 3A,and proinflammatory factors such as IL-1?, IL-6,IL-8 and TNF-?,in peripheral blood monocytes/macrophages(MC).The expression of TLR4 protein in MC was analyzed by flow cytometry.Results①Compared with the control group,the mRNA levels of TLR4, MD-2,MyD88,IRAK-4 and TRAF6 in KD group were up-regulated significantly(P＜0.01),and the expression level of TLR4 protein was also found to be up-regulated in KD group during acute phase.It was detected that expression levels of TLR4 protein in KD with coronary artery lesion(KD-CAL~+)was significantly higher than that of KD without coronary artery lesion(KD-CAL-)[flow cytometry:(6.5?1.7)% vs(11.9_+2.4)%,P＜0.01].②The expression level of negative-regulatory factors such as IRAK-M and Triad3A were significantly up-regulat- ed in acute phase of Kawasaki disease,while the mRNA levels of IRAK-M and Triad3A in KD-CAL~+ group was found to be significantly lower than those of KD-CAL~- group(P＜0.01).No difference of T1/ST2 mRNA expres sion level was detected among all groups(P＞0.05).③The expressions of proinflammatory eytokines such as IL-1?, IL-6,IL-8 and TNF-?in monoeytes/macrophages during acute phase of Kawasaki disease were higher than those of the control group(P＜0.01),and expression of proinflammatory cytokines in KD-CAL~+ group was significantly higher than that of KD-CAL~- group.Conclusion Relative insufficient expression of negative-regulatory factors, such as IRAK-M and Triad3A,maybe correlate with immunological pathogenesis of Kawasaki disease.

A strain with 2 g/L yield of poly-?-lysine was firstly screened from soil on an agar plate containing methylene blue and then further selected by a flask experiment. The strain was preliminarily identified as Streptomyces albulus. The experiment about medium optimization showed that glucose was the best carbon sources and yeast extract and (NH_4)_2SO_4 were the best compound nitrogen sources. The yield of ?-PL reached 3.9 g/L with optimal medium.

Objective To analyze the etiologic factors and clinical manifestation of intraventricular hemorrhage (IVH) in preterm infants.Methods One hundred and seventy-two preterm infants from June 2005 to August 2006 were accrued to investigate their gestational age,birth weight,birth history,and clinical symptoms.Cranial chromatic ultrasound was used to scan the preterm infants and diagnose IVH.Results 1.The incidence of IVH was associated with gestational age (?2=6.40 P=0.011);2.The incidence of IVH was also associated with birth weight(?2=26.49 P=0);3.IVH usually occurred within 72 h with mild clinical manifestations and was diagnosed within 5 days after birth;4.IVH occurred more frequently and more severe in infants with severe asphyxia than those with mild asphyxia.Conclusions Early gestational age,low birth weight, and severe asphyxia are risk factors for IVH.The clinical symptoms of IVH are usually mild in most patients.Cranial chromatic ultrasound is a reliable,sensitive and convenient method of detection for IVH in preterm infants.

OBJECTIVETo study the active constituents of Ranunculus ternatus.

METHODThe constituents were isolated with silica gel and Sephadex LH -20 gel column chromatography and purified by HPLC. Their structures were elucidated by spectroscopy.

RESULTSixteen compounds were obtained and identified as Stigmasta4, 6, 8 (14), 22-tetraen-3-one (1), 5-hydroxymethyl furaldehyde (2), gamma-keto-delta-valerolactone (3), pantolactone (4), 5-hydroxymethyl-dihydro-furan-2-one (5), methyl 5-hydroxy-4-oxopentanoate (6), methyl hydrogen succinate (7), succinic acid monoethyl ester (8), 3, 4-dihydroxybenzoic acid methyl ester (9), p-hydroxycinnamic acid (10), 4-oxo-pentanoic acid (11), succinic acid (12), nonanedioic acid (13), 4-hydroxybenzoic acid (14), 4-hydroxybenzaldehyde (15), stigmasterol-3-O-beta-D-glucopyranoside (16).

CONCLUSIONCompound 1, 4 -15 are obtained from this plant for the first time.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Ranunculus ; chemistry

OBJECTIVEImmunosuppressant tacrolimus (FK506) has shown neuroprotective effects on hypoxic-ischemic brain damage (HIBD) in the adult animal model. This study investigated whether FK506 has a protection against HIBD in neonatal rats by examining growthjassociated protein-43 (GAP-43) expression in the hippocampus.

METHODSNinety-six seven-day-old Sprague-Dawley rats were randomly divided into three groups: sham-operation, HIBD and FK506 intervention group. HIBD was induced in the later two groups. The FK506 intervention group was intraperitoneally injected with FK506 immediately after HIBD, at a dosage of 1 mg/kg daily, for three days. The HIBD group was injected with normal saline. Immunohistochemical technical was applied to examine GAP-43 expression in the hippocampus 24 and 72 hrs and 7 and 14 days after HIBD.

RESULTSCompared with the HIBD group, hematoxylin-eosin staining showed attenuated neuronal necrosis in the FK506 intervention group. In the HIBD group, the expression of GAP-43 increased significantly 72 hrs, and 7 and 14 days after HIBD compared with that in the sham-operation group. The GAP-43 expression in the FK506 intervention group was significantly higher than that in the HIBD group 72 hrs and 7 days after HIBD.

CONCLUSIONSFK506 might have neuroprotective effects against HIBD in neonatal rats.

Animals ; Animals, Newborn ; GAP-43 Protein ; analysis ; Hippocampus ; chemistry ; drug effects ; Hypoxia-Ischemia, Brain ; drug therapy ; metabolism ; pathology ; Immunosuppressive Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Tacrolimus ; pharmacology

Sini decoction (SND), a classical traditional Chinese medicine emergency formula recorded in Shanghan Lun (Treatise on Febrile Diseases), which is composed of Aconiti Lateralis Preparata Radix, Glycyrrhizae Preparata Radix and Zingiberis Rhizoma. Modern clinical and pharmacological researches have shown that SND can protect the myocardium effectively during myocardial ischemia reperfusion injury (MI-RI). A myocardial ischemia reperfusion injury model of H9c2 cardiomyocytes in vitro has been established. Four groups, control group, MI-RI Model group, SND group and SND without Glycyrrhizae Radix group, were arranged. The livability, the level of LDH and CK activity in H9c2 cardiomyocytes in different groups were tested. By combining with principal components analysis (PCA), partial least squares discriminant analysis (PLS-DA), orthogonal partial least squares projection of latent structures-discriminant analysis (OPLS-DA), 17 biomarkers in extracellular fluid were identified and 15 of them were related to the pathway of biological processes. The results showed that the attenuate and synergistic mechanism between Aconiti Lateralis Praeparata Radix and Glycyrrhizae Radix for toxicity reduction was related with the glycolysis, lipid metabolism, citrate cycle and nitrogen metabolism of amino acids metabolism. The study proved the effect on H9c2 cardiomyocyte treated by MI-RI injury both SND group and SND without Glycyrrhizae Radix group, and compared with the SND without Glycyrrhizae Radix, the protective effect of myocardial ischemia reperfusion injury model of H9c2 cardiomyocyte from SND was stronger.
Aconitum ; chemistry ; Animals ; Drug Synergism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Glycyrrhiza ; chemistry ; Humans ; Male ; Metabolomics ; Myocardial Reperfusion Injury ; drug therapy ; genetics ; metabolism ; Myocytes, Cardiac ; drug effects ; Rats, Sprague-Dawley