Adolescent ; Adult ; Aged ; Carcinoma, Papillary ; genetics ; metabolism ; pathology ; Child ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; PTEN Phosphohydrolase ; genetics ; metabolism ; Promoter Regions, Genetic ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; Young Adult

In present study,bovine Lactoferricin was first secretly expressed in Pichia pastors yeast expression system.The synthesized LfcinB gene fragment was cloned into expression vector pPIC9K,and then obtained recombinant plasmid,designated as pPIC9K-LfcinB,was linearized and transformed into Pichia pastors strains SMD1168 by electroporation.The transformants were screened with Geneticin and multiply-copy colonies were harvested,in which LfcinB gene was verified to inserted into yeast chromosome stably.The positive recombinant Pichia strains were induced with methanol to express LfcinB in culture supernatant.It's expressive products has high activity of killing bacteria.We concluded that LfcinB gene was cloned and integrated into yeast chromosomes,and obtained expression peptide was tested to have high antibacterial activity.

Objective To evaluate the efficacy and safety of a new drug,human urinary kallidinogenase,against acute brain infarction.Method A 15-center,randomized,double-blinded and 3:1 placebo-controlled study was carried out.Acute brain infarction within 48 hours of onset in the territory of the middle cerebral artery were indicated as subjects;kallidinogenase or placebo which was dissolved in 50 ml saline,was slowly injected intraveousely within 30 minutes daily for 3 weeks.The European Stroke Scale and Barthel Index were used to evaluate the neurological deficit and the activities of daily living(ADL),followed by a follow-up at the end of the third month.Results 446 patients were enrolled,who completed ITT analysis,including 330 in kallidinogenase group and 116 in placebo group,meanwhile 421 proceeded with PP analysis(311 and 110 respectively).There were no significant differences of the baseline data between the 2 groups.At the end of treatment,the ESS scores increased by 55.1%?33.0% and 44.7%?32.8% respectively in kallidinogenase group(KG)and placebo group(PG,P=0.0022),the difference being significant.PP analysis had similar results.As for ADL,follow-up 90 days after the treatment showed 374 cases followed,280 in KG and 94 in PG;1 died in PG,while none in KG.In KG,the cases whose BI≥50 were significantly more than those in PG(P=0.0228).Adverse events possibly or definitely attributable to the drug were observed in 27 cases(7.74%),mostly were mild,such as palpitation,flush,dizziness, nausea etc,without special management needed.Only 2 died which was confirmed not correlated to kallidinogenase,and another 2 cases of sudden blood pressure drop were observed.The blood pressure drop, quickly restoring soon after the withdrawal of kallidinogenase and use of hemopiesic drugs,was considered to be caused by the combination use of anti-hypertensive drug ACEI and quick infusion speed.Conclusion Kallidinogenase is efficacious for acute brain infarction in improving the neurological deficits,which is safe in clinical use.

A HPLC method has been developed in the current investigation for simultaneous determination of three chemical markers of by akangelicin, imperatorin and isoimperatorin in Radix Angelicae Dahuricae. Separation was performed at 30 degrees C. on achromatographic column of Platisil ODS C18 (4.6 mm x 250 mm, 5 μm). The mobile phase was acetonitrile-water. The flow rate was 1.0 mL x min(-1) and the detection wavelength was 254 nm. The results showed that the three chemical markers could be well resolved and that in the selected linear range, all calibration curves of the three chemical markers showed good linearity (R2 ≥ 0.999 8). The recoveries of byakangelicin, imperatorin and isoimperatorin were 100.83%, 100.10% and 103.52%, respectively, and RSD were 1.7%, 0.77% and 0.41% (n = 6), respectively. The data suggested that the developed HPLC method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Radix Angelicae Dahuricae. Applications of this method showed that the three chemical markers had higher contents in the Bozhou Anhui and Changge Henan than others.
Angelica ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Furocoumarins ; analysis ; Plant Roots ; chemistry ; Quality Control

OBJECTIVETo study the effect of sodium ferulate (SF), an active component of Radix Angelica, on lung damage induced by ozone (03).

METHODSMice model of lung injury was induced by ozone inhalation and treated with SF. The level of lipid peroxide and microviscosity in alveolar epithelial cell membrane of the mice was determined, and the structural change of lung cells was observed by microscopy.

RESULTSOzone could increase the level of malondialdehyde (MDA) and the microviscosity in alveolar epithelial cell membrane, and induce inflammatory changes in morphologic structure. These abnormal changes were improved after SF administration, which was manifested as alleviation of heightened microviscosity, increase of membrane fluidity, as well as the basically normalized pulmonary cellular structure under microscope.

CONCLUSIONSF has a preventive effect against oxidized pulmonary injury induced by ozone, the action of which could be through scavenging oxygen free radicals, reducing lipid peroxide production, increasing membranous fluidity and mitigating inflammatory changes in cell structure. sodium ferulate, ozone, malondialdehyde, membranous fluidity, morphology

Animals ; Cell Membrane ; drug effects ; Coumaric Acids ; pharmacology ; Female ; Free Radical Scavengers ; pharmacology ; Lung ; drug effects ; metabolism ; pathology ; Male ; Malondialdehyde ; analysis ; Membrane Fluidity ; drug effects ; Mice ; Mice, Inbred ICR ; Ozone ; toxicity

Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Fibroma ; metabolism ; pathology ; surgery ; Humans ; Keratins ; metabolism ; Middle Aged ; Vimentin ; metabolism

Objective To observe the effect of T-2 toxin on growth and development of rat epiphyseal plate of left knee and metaphyseal bone of femur and tibia in normal and low nutritional status, to find out possible pathogenic factors of Kashin-Beck disease and provide experimental basis for early intervention. Methods Ninety 3-week-old Wistar rats, weighing 60 - 70 g, were randomly divided into three groups: control group(general feed), T-2 toxin + general feed group, T-2 toxin + low nutrition feed group, thirty rats in each group with equally sex ratio. T-2 toxin (1.0 mg/kg) was administered orally 5 times a week via a gavage needle for 4 weeks. The change of hair, activity and body weight was observed. After 1, 2, 4 weeks, the epiphyseal plate of left knee and metaphyseal bone of femur and tibia (including distal femur and proximal tibia) were collected. Specimens were processed with HE and Masson staining. The morphology of chondrocytes and matrix collagen content in epiphyseal plate was observed. Trabecular bone volume fraction in tibial metaphyseal bone was analyzed by Image-Pro Plus 6.0 software. Results In the control group, rats were in good movement and hair with light, but in T-2 toxin + general feed group and T-2 toxin + low nutrition feed group, rats were found with reduced activities and hair with dark color. Body weights(g) of the control group, the T-2 toxin + general feed group and the T-2 toxin + low nutrition feed group were 81.0 ± 6.2, 79.0 ±5.1, 77.0 ± 7.5, respectively, by the end of first week; 101.8 ± 6.7, 97.0 ± 6.8, 93.0 ± 5.3, respectively, by the end of second week; 151.1 ± 15.7, 126.5 ± 11.9, 106.5 ± 11.5, respectively, by the end of fourth week. There was significant difference in groups by second week and the fourth week (F = 9.72, 41.65, all P ＜ 0.05 ). There was significant difference among multi-groups by the fourth week(all P ＜ 0.01 ). Under light microscope, at the second weeks, coagulative necrosis of chondrocytes was found in hypertrophic zone in the two groups with T-2 toxin; at the fourth weeks, cell necrosis increased. Masson staining showed collagen staining in the two groups with T-2 toxin significantly turned to clear pale coloration, indicating that the collagen matrix was significantly reduced. Image analysis showed there was significant difference in groups at the second and fourth week(F= 9.72, 41.65, all P＜ 0.05)in tibial metaphyseal trabecular bone volume fraction. There was significant difference between T-2 toxin + low nutrition feed group[(0.55 ± 0.12)％, (0.21 ± 0.0)％] and control group[(0.67 ± 0.09)％, (0.51 ± 0.14)％] by the second and fourth week(all P ＜ 0.01 ). Conclusions Under normal nutritional status, T-2 toxin can induce hypertrophic epiphyseal cartilage necrosis, collagen content decreased in epiphyseal plate, metaphyseal trabecular bone formation disorders; in the low nutritional status, T-2 toxin can lead to rat epiphyseal necrosis and significant metaphyseal bone disorder, but whether the performance is related to Kaschin-Beck disease needs to be studied further.

OBJECTIVE: To investigate the regulation of hepatitis C virus (HCV) core protein on the activity of signal transducers and activators of transcription 3 (stat3). METHODS: A cell line expressing stable HCV core protein-QSG7701-core was constructed by transfecting the pcDNA3. 1-core (expressing HCV core protein) into the human immortalized hepatocyte line QSG7701. The phosphorylation and DNA binding activity of stat3 were detected by immunocytochemistry, Western blot, and electrophoretic mobility shift assay (EMSA). RESULTS: The expression level of phosphorylated stat3 in QSG7701-core cells was significantly lower than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells, but there were no significant differences in the expression levels of total stat3 among the 3 groups. The positive signal of phosphorylated stat3 in nucleus of QSG7701-core cells was obviously weaker than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells. EMSA showed that DNA binding activity of stat3 in QSG7701-core cells significantly decreased. CONCLUSION The expressionof HCV core protein in human hepatocyte line may suppress the phosphorylation and DNA binding activity of stat3, which may be one of the causes for resistance against interferon.
Cell Line ; DNA-Binding Proteins ; metabolism ; Hepatocytes ; cytology ; metabolism ; Humans ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Transfection ; Viral Core Proteins ; biosynthesis ; genetics

OBJECTIVE: To explore the changes of apoptosis of cells in the lung tissue of rats with silica instillation and to its significance in silicosis, and to clarify the role of caspase-3 in the apoptosis progress. METHODS: Forty-eight rats were randomly divided into saline control groups and silica instillation groups, and the silicosis model was established in rats. Flow cytometry was used for detecting the rate of apoptosis at various stages. Immunohistochemistry for the expression of cleaved caspase-3. RESULTS: The model of rat silicosis was established successfully. The apoptosis rate in the experimental group was significantly higher than that in the control group, and was increased with time. Caspase-3 was mainly expressed in alveolar epithelium cells, pulmonary macrophages and infiltrated inflammation cells. The expression of caspase-3 in the experimental group was stronger than that in the control group, but its expression intensity was not related to the cell apoptosis (r = 0.215, P > 0.05). CONCLUSION The apoptosis of the lung cells plays an important role during rat silicosis genesis. Caspase-3 plays an important role in regulating cell apoptosis during rat silicosis genesis.
Animals ; Apoptosis ; physiology ; Caspase 3 ; Caspases ; metabolism ; Lung ; enzymology ; pathology ; Male ; Pulmonary Fibrosis ; etiology ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Silicosis ; enzymology ; pathology

A fragment containing amino acid residues 561～578 of HSV-2 glycoprotein G(gG2) was obtained by PCR assembling technique,and doubly cloned into vector pET-KDO.The recombinant plasmid was transformed to BL21(DE3)plysS.Fusion protein,of molecular weight about 39kDa was highly expressed by induction of IPTG.Western blot result showed the fusion protein had good antigenicity.After putification and digestion,the purity reached 95%.The digested purified protein was analysed by ELISA and showed good sensitivity and specificity.The recombinant protein should be useful for type-specific serodiagnosis of HSV-2.