Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of vascular endothelial growth factor (VEGF) in human hepatic cancer cell line Hep G2.Methods Cultured Hep G2 cells were treated by AngⅡ with various concentrations and were collected at different time points.Then total RNA was extracted.The expression of VEGF mRNA in cultured Hep G2 was determined by relative quantitative reverse transcription polymerase chain reaction(RT-PCR),and the proliferation was detected by methyl thiazolyl tetrazolium (MTT).Results AngⅡ stimulated the proliferation of HepG2 cells,and enhanced the expression of VEGF mRNA (P

Chloroplast-based expression system is promising for the hyper-expression of plant-derived recombinant therapeutic proteins and vaccines. To verify the feasibility of obtaining high-level expression of the SARS subunit vaccine and to provide a suitable plant-derived vaccine production platform against the severe acute respiratory syndrome coronavirus (SARS-CoV), a 193-amino acid fragment of SARS CoV spike protein receptor-binding domain (RBD), fused with the peptide vector cholera toxin B subunit (CTB), was expressed in tobacco chloroplasts. Codon-optimized CTB-RBD sequence was integrated into the chloroplast genome and homoplasmy was obtained, as confirmed by PCR and Southern blot analysis. Western blot showed expression of the recombinant fusion protein mostly in soluble monomeric form. Quantification of the recombinant fusion protein CTB-RBD was conducted by ELISA analysis from the transplastomic leaves at different developmental stages, attachment positions and time points in a day and the different expression levels of the CTB-RBD were observed with the highest expression of 10.2% total soluble protein obtained from mature transplastomic leaves. Taken together, our results demonstrate the feasibility of highly expressing SARS subunit vaccine RBD, indicating its potential in subsequent development of a plant-derived recombinant subunit vaccine and reagents production for antibody detection in SARS serological tests.
Chloroplasts ; metabolism ; Cholera Toxin ; Protein Interaction Domains and Motifs ; Recombinant Fusion Proteins ; biosynthesis ; SARS Virus ; Spike Glycoprotein, Coronavirus ; biosynthesis ; Tobacco ; metabolism ; Vaccines, Subunit ; biosynthesis

Objective To observe the effects of high mechanical index ultrasound contrast on hepatic metastasis of colorectal cancer. Methods Hepatic metastasis models were established by injecting Lovo cells into the spleen of SD rats.This study was divided into control group, microbubble plus ultrasound radiation group simple contrast agent group and simple ultrasound radiation group.Ultrasound contrast agent SonoVue(1 ml/kg) was injected via tail vein, high mechanical index ultrasound contrast (frequency 1.5 MHz,MI 1.7) was performed intermittently for 2 min.After 10 d,the animals were killed and the size and number of hepatic metastasis were compared.Histological pathology and its ultrastructure were observed.Results There were no significant differences between contral group,simple ultrasound radiation group and simple contrast agent group in metastatic rate ( P＞0.05).There were no obvious differences between contral group, simple ultrasound radiation group and simple contrast agent group in cells'ultrastructure by transmission electron microscopy (TEM).The number and size of hepatic metastasis decreased obviously in microbabble plus ultrasound radiation group ( P＜0.01 ), the mitochondrion was swelling and the gaps of endothelial cells were loose. Conclusions High mechanical index ultrasound contrast can inhibit the hepatic metastasis of colorectal cancer.

Objective To assess the value of ultrasound examination in transvaginal diagnosis of normal fetal heart with 11-14 weeks of gestation.Methods Totally 158 cases of normal fetal heart with high risk pregnancy and nuchal translucency thickness were examined by two ultrasoud approaches:transvaginal(TVS) and tranabdomina(TAS) in 11-14 weeks of gestation.Results The group of TVS was obviously clearer than TAS by displaying the normal fetal cardiac structural in 12 week of gestation and 13-14 weeks of gestation with significant difference between the two groups (P<0.05),respectively.No significant difference between the two groups in 11 week of gestation was observed.Conclusion The transvaginal echocardiogram is of clinical value in the high risk gravida during the late first and the early second trimester.

Objective To Study the effects of high mechanical index ultrasound radiation microbubble on colon cancer cell skeletal.Methods Lovo cell were cultured in vitro,and this study was divided into control group,simple contrast agent group,microbubble plus ultrasound radiation group and simple ultrasound radiation group.Ultrasound contrast agent SonoVue and ultransound (frequency 1.5 MHz and MI 1.7) were used.Results In the control group,the microtubes were densest in dyeing silk,and extended to the edge of a cell.In the ultrasound and microbubble group,the microtubes were weaker and thin,the.network was mainly in the structure of the long axis.The cells arranged in a slight pigmentation to the density cell,to reach out many fine short hair,there were obviously a sense of silk and directional.In the microbubble and ultrasound group,Lovo cells streamed of the central significantly reduced,fluorescent lighting,with short hair.There was no difference in the ultrasound group and the microbubble group with microfilament and microtubule.Conclusions High mechanical index ultrasound radiation can changed into vacant,a trace of the assembly and distributed to the tumor cells are attacking,the transfer of restraining.

Advances in mechanism and application of the heating effect in breeding of microorganism are reviewed in this paper. Heat produces mutagenesis effect and screening effect. Heating mutagenesis effect is occurred through the substitution of G-C base pair induced by heat, and heating screening effect produces higher forward mutation rate induced by other mutagens.

OBJECTIVETo evaluate the efficacy and safety of Xinmailong Injection (XI) in treatment of chronic heart failure (CHF) patients with qi-yang deficiency and blood stasis resistance syndrome (QY-DBSRS).

METHODSTotally 238 CHF patients with QYDBSRS were assigned to the treatment group (118 cases) and the control group (120 cases) by randomized, double-blind, placebo parallel controlled method. Patients in the treatment group received routine therapy and XI (100 mg/2 mL, by dripping at 5 mg/kg, twice per day for 5 consecutive days), while those in the control group received routine therapy and XI mimetic agent (100 mg/2 mL, by dripping at 5 mg/kg, twice per day for 5 consecutive days). The heart function classification of New York Heart Association (NYHA), 6-min walking distance, left ventricular ejection fraction (LVEF), scores for Chinese medical symptoms were observed before and after treatment, and safety assessed.

RESULTSTotally 235 patients actually entered full analysis set (FAS), including 120 cases in the control group and 115 cases in the treatment group. The total effective rate of heart function, 6-min walking distance and increased post-pre-treatment distance in the experimental group were superior to those of the control group with statistical difference (all P < 0.05). Compared with the control group, increased value of post-pre-treatment LVEF, the total effective rate of Chinese medical syndrome efficacy, scores for Chinese medical symptoms and decreased post-pre-treatment value of Chinese medical syndrome scores were obviously improved (all P < 0.05). There was no significant difference in the incidence of adverse events between the two groups (P > 0.05).

CONCLUSIONSXI could improve the heart function of CHF patients, improve Chinese medical symptoms, elevate exercise tolerance, and improve LVEF. It had no obvious toxic and side effects.

Chronic Disease ; Double-Blind Method ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Heart Failure ; drug therapy ; Humans ; Injections ; Medicine, Chinese Traditional ; Phytotherapy ; Qi ; Syndrome ; Yang Deficiency

OBJECTIVETo increase the recovery rate of ethyl acetate after extracting tripterygium wifordii extractum and to decrease product cost.

METHODAfter extracting tripterygium wifordii extractum with ethyl acetate, 3 times saturated salt water was added in it so as to recovery ethyl acetate distilled under normal atmospheric pressure. Ethyl acetate containing salt water was purified through Na2SO4 column.

RESULTEthyl acetate purified could be used repeatedly and the recovery rate was up to 85%.

CONCLUSIONThis method is completely adapted for mass production.

Acetates ; Drugs, Chinese Herbal ; isolation & purification ; Plants, Medicinal ; chemistry ; Sodium Chloride ; Technology, Pharmaceutical ; economics ; methods ; Tripterygium ; chemistry

In part of the patients with blood disease or malignant tumors, especially those with leukemia and multiple myeloma, the disease state and unsuitable treatment often resulted in the inconsistence between positive and negative ABO blood group, displaying attenuation of the antigen or antibody of ABO blood group. This study was purposed to analyze the course of inconsistence between positive and negative ABO blood group and to perform the correct typing of erythrocytes and genes. The serology, absorption and elution test were used to examine the 12 tumor patient of the inconsistence between positive and negative typing. The 6th, 7th exon and 5-7th introns were amplified by PCR for questionable samples, and the gene sequencing of exon was performed. The results showed that 9 specimens were determined as 6 of A group, 2 of O group, 1 of B group, 3 cases were identified as O46, B108, and A102 group, respectively, by the serology, absorption and elution typing. The genotype of 2 cases among them was not identified because of the erroneous PCR amplified result or the contradicted sequencing results, failing to determine the ABO genotype. It is concluded that the serological method for blood grouping, genotyping, absorption and elution method can be used for the blood samples unable to typing because of the inconsistence between positive and negative typing of ABO group, therefore, guaranteeing the safety and effectiveness.
ABO Blood-Group System ; genetics ; immunology ; Blood Grouping and Crossmatching ; methods ; Genotype ; Genotyping Techniques ; methods ; Humans ; Neoplasms ; genetics ; immunology

OBJECTIVETo adapt the candidate strains of hemorrhagic fever with renal syndrome (HFRS) purified vaccine to Vero cells and to study their antigenicity and immunogenicity.

METHODSThe viral strains H8207 (Hantaan virus, HTN) and Y86013 (Seoul virus, SEO) were continuously propagated in Vero cell by the terminal dilution method and studied the characteristics of virus multiplication, viral titers and the amounts of virus antigen after serial passages. Three batches of crude monovalent inactivated vaccine were developed using the different passages of these 2 viral strains.

RESULTSThe strains H8207 and Y86013 adapted to Vero cells and stably grew on the cells with high titers. Rabbits immunized with the crude vaccines of H8207 and Y86013 showed 100% sero-conversion and the neutralizing antibody titers of the rabbit immune sera reached 1?10 at 4 weeks after 2 times of immunization.

CONCLUSIONSThe results suggest that these 2 candidate strains had adapted to Vero cells, possessed high titers and good immunogenicity and be feasible to prepare the HFRS purified vaccine in Vero cells.

Animals ; Antibodies, Viral ; blood ; Cercopithecus aethiops ; Hantaan virus ; growth & development ; immunology ; Hemorrhagic Fever with Renal Syndrome ; prevention & control ; Mice ; Neutralization Tests ; Rabbits ; Seoul virus ; growth & development ; immunology ; Vaccination ; Vaccines, Inactivated ; immunology ; Vero Cells ; Viral Vaccines ; biosynthesis ; immunology