The intracellular hepatitis B surface antigen (HBsAg) content per cell was increased by 7.2-fold in the culture with 1.5% dimethyl sulfoxide (DMSO) compared with that in the control without DMSO, while the extracellular HBsAg production and specific productivity were only improved by 70% and 3.2-fold, respectively. Electron microscope has been employed to reveal large dilated structures within recombinant CHO cells in the presence DMSO. The dilated structures have a distribution within whole cytoplasm, and some dilated areas were engulfed in the nucleus. These large, dilated structures were not observed in the control. Immunogold labeling was used to discover the accumulated HBsAg was localized within these dilated areas, and some HBsAg-specific labels were detected in the nucleus membrane, owing to the encroachment of the dilated areas upon nucleus. The result could help to reveal the mechanism of intracellular HBsAg accumulation in the presence of DMSO.

Quantitative NMR (qNMR) is a technology based on the principle of NMR. This technology does not need the references of the determined components, which supplies a solution for the problem of reference scarcity in the quantitative analysis of traditional Chinese medicines. Moreover, this technology has the advantages of easy operation, non-destructiveness for the determined sample, high accuracy and repeatability, in comparison with HPLC, LC-MS and GC-MS. NMR technology has achieved quantum leap in sensitivity and accuracy with the development of NMR hardware. In addition, the choice of appropriate experimental parameters of the pre-treatment and measurement procedure as well as the post-acquisition processing is also important for obtaining high-quality and reproducible NMR spectra. This review summarizes the principle of qHNMR, the various experimental parameters affecting the accuracy and the precision of qHNMR, such as signal to noise ratio, relaxation delay, pulse width, acquisition time, window function, phase correction and baseline correction, and their corresponding optimized methods. Moreover, the application of qHNMR in the fields of quantitation of single or multi-components of traditional Chinese medicines, the purity detection of references, and the quality analysis of foods has been discussed. In addition, the existing questions and the future application prospects of qNMR in natural product areas are also presented.
Biological Products ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; methods ; Medicine, Chinese Traditional ; Reproducibility of Results

Argyrogramma agnata has been selected as a substitutive host insect for producing Dendrolimus punctatus Cytoplasmic Polyhedrosis Virus (DpCPV). In our experiment, it is very susceptible to DpCPV. The DpCPV produced in A. agnata is designated Aa-DpCPV. The cytoplasmic polyhedra body (CPB), the virion size and the shape of Aa-DpCPV are same as that of its original DpCPV (DpCPV-W1984). The RNA bands of Aa-DpCPV and DpCPV-W1984 all have 10 RNA segments respectively in 3% PAGE, which molecular weights ranged in size from 2.98×106 to 0.66×106 Dalton. Aa-DpCPV has the same strong toxicity as that of DpCPV-W1984 (from D. punctatus) to D. punctatus (Walker) larva. So it can be applied to the pine caterpillar control. The DpCPV yield in A.agnata is 2.5×108CPB/larva.

Adenocarcinoma, Follicular ; pathology ; physiopathology ; Female ; Humans ; Kidney Diseases ; pathology ; physiopathology ; Kidney Neoplasms ; physiopathology ; Thyroid Neoplasms ; pathology ; physiopathology ; Young Adult

Objective To detect the activity of transglutaminase1(TGM1)and gene mutation in a family with lamellar ichthyosis.Methods Immunohistochemistry technique was used to detect the activity of transglutaminase1.Complete encoding sequences of TGM1gene were analyzed in this family by using PCR-DNA sequencing.Results No activity of transglutaminase1was detected in the proband's skin.A nonsense mutation of C604T located in exon4of TGM1gene was identified by PCR-DNA sequencing,which caused a premature termination of Q202X and a defective polypeptide truncated by615amino acids in C-terminus.A heterozygous C604T mutation was carried by both of the proband' s parents.Conclusions The proband of lamellar ichthyosis in this family shows loss of transglutaminase1activity,which is resulted from a truncated transglutaminase1coded by the homozygous mutant TGM1gene.

Tongluo Xingnao effervescent tablet (TLXNET) is a patented prescription, which comes from modified Xionggui decoction and can improve cognitive function. However, its effect on the urine metabolites and anti-dementia mechanism in the dementia model rats induced by hippocampal injection with Aβ25-35 remains unclear. The experiment focused on the changes in trajectory and inter-relationship among the urinary metabolite of rats in the blank group, Aβ25-35 hippocampal injection dementia model group and the TLXNET intervention group, in order to determine theirs characteristic metabolic markers and explain the anti-dementia effect of TLX-NET base on the change of metabolic trajectory of these bio-markers. According to the experimental results, 5, 6-indolequinone, 4-hydroxyphenyl pyruvic acid (4-HPPA), cortisol and 3-thiosulfate lactic were preliminarily identified as the characteristic metabolic markers. They mainly participate in dopamine system, glucocorticoids and energy metabolic pathways. TLXNET can apparently downregulate the disturbances of metabolic trajectory of the four bio-markers. The experiment indicates that the dementia model induced by injecting Aβ25-3 into hippocampus has its characteristic endogenous metabolic markers in urine, and ELXNET can ameliorate dementia by down-regulating the disturbances of metabolic trajectory.
Amyloid beta-Peptides ; metabolism ; toxicity ; Animals ; Biomarkers ; urine ; Dementia ; drug therapy ; urine ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hippocampus ; drug effects ; metabolism ; Humans ; Male ; Metabolomics ; Peptide Fragments ; metabolism ; toxicity ; Rats ; Rats, Sprague-Dawley ; Tablets ; administration & dosage ; Urine ; chemistry

Twenty healthy subjects in each of 3 groups were fed with monounsaturated fatty-acid diet,polyunsaturated fatty-acid diet, or saturated fatty-acid diet separately for 3 days. It suggested that monounsaturated fatty-acids may ameliorate the oxidative stress and improve insulin sensitivity.

Objective To evaluate short-term curative effect of endovascular abdominal aortic aneurysm repair. Methods Twelve cases of infra-renal abdominal aortic aneurysms were checked. Results The av-erage bleeding in the operation was 245 millimeter, the avenage hospitalization time was 8.6 days, and the average abrosia time was 1.5 days. White blood cell, hemoglobin, thrombocyte, hepatic function, and renal function were in the normal limits. Prothrombin time and activated partial thromboplastin time were post-poned after operation, and recovered normally within one week. Complications of operations were as follows:1 case of pulmonary infection, 2 cases of abdominal distention, and 1 case of intraoperafive endoleak. The former two kinds of complications alleviated after conventional treatment, and the latter disappeared naturally after 3 months. Conclusion Endovascular abdominal aortic aneurysm repair is safe, mini-invasive, and has little disturbance for body internal environment.

Targeting induced local lesions in genomes (TILLING) is a reverse genetics method for functional genomics research.It is possible to screen for point mutations in the populations of EMS mutagenesis with highthroughput and lowcost. EcoTILLING a method based on TILLING ,was developed for detecting multiple types of polymorphisms in germplasm collections,such as single nucleotide polymorphism,small deletion and insertion etc.Rice is a very important food crop and a model plant for genome research also. There are complete genome sequence and a lot of other bioinformatics resources about it.So the markerassisted breeding is becoming more and more important in rice breeding. Some issues based on TILLING about identifying germplasm based on gene sequence,EMS mutagenesis breeding,developing functional marker in rice breeding in future were discussed.

OBJECTIVETo construct a chimeric SEA-hPLAP-1 cDNA with gene splicing by overlap extension.

METHODSThe SEA gene and a DNA fragment encoding the signal for GPI-anchor attachment of hPLAP -1 were amplified by PCR. The two amplified gene sequence was annealed to form a chimeric GPI- anchored SEA molecule with gene splicing by overlap extension. The resulting chimera was cloned in pGEM-T vector and verified by sequencing analysis.

RESULTA chimeric SEA-hPLAP-1 cDNA was successfully constructed with gene splicing by overlap extension.

CONCLUSIONGene splicing by overlap extension is a successful specific PCR technique for gene recombination.

Alkaline Phosphatase ; Base Sequence ; Enterotoxins ; genetics ; GPI-Linked Proteins ; Isoenzymes ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA Splicing ; Recombinant Fusion Proteins ; genetics