The yeast fusant ZFF-28, which is high in biomass production and rich in selenium, was constructed after mutagenesis and protoplasts fusion between yeast strains. The total selenium content of ZFF-28 is 1.8 and 1.0 times higher than that of the parental strains Saccharomyces cerevisiae ZY-67 and Saccharomyces kluyveri SZY-198 respectively. Using single factor tests and a L16(4(3) x 2(1)) orthogonal design, the cultivation conditions was optimized as: 50mL culture in 250mL shake flasks in molasses containing 6% sugar and 60microg/mL Se at 28 degree C for 25h at 220 r/min, with the initial pH adjusted to 6.0 - 6.5. Under the optimized conditions, the biomass (dry weight) reached 8.2g/L and the Se content of the cells reached 2050microg/g, with organic and inorganic Se contents being 91% and 9% respectively.
Biomass ; Hydrogen-Ion Concentration ; Saccharomyces ; genetics ; growth & development ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism ; Selenium ; metabolism ; Selenium Compounds ; metabolism

Adult ; Aged ; Aged, 80 and over ; Female ; Head and Neck Neoplasms ; surgery ; Humans ; Intubation, Intratracheal ; methods ; Laryngoscopes ; Male ; Middle Aged

Objective To evaluate the potential relationship between isoforms of BRCA1 associated RING domain 1 ( BARD1 ) and the pathophysiologic markers of sporadic breast carcinoma of female Han ethnic group. Methods The expression of BARD1 isoforms in 39 breast carcinomatous tissue, 12paracancerous-normal breast tissue and 7 controlled normal breast tissue was detected by reverse transcription polymerase chain reaction (RT-PCR) and then cloned and sequenced.The difference of isoforms expression and their clinical significance were analyzed. Results There were four transcriptive products of BARD1 found in all these candidates,named full lenth,isoform γ,isoform δ and isoform e.The positive rate of isoform γ and δ was higher in carcinomatous tissues than in paracancerous-normal tissues and normal breast tissue in healthy women ( P ＜ 0.05 ). Carcinomatous tissue expressed more kinds of isoforms.There was significant difference between carcinomatous tissue and paranormal/nomal tissue ( P =0.0075 ).There was significant correlation between isoform ε positive and poor prognosis factors such as poorly differentiation,HER2 positive,poor pathologic type and larger breast cancer lumps(P ＜ 0.05 ). Conclusions There are significant differences in the expression of BARD1 isoforms among different kinds breast tissues in the female Han ethnic group.Positive isoform ε may predict poor prognosis of breast carcinoma in the female Han ethnic group.

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment

OBJECTIVETo investigate the chemical constituents of Epimedium brevicornum.

METHODThe chemical constituents were isolated by using silica gel column chromatography and preparative TLC. The structures were identified on the basis of physical-chemical constants and spectral data.

RESULTFive compounds were isolated and identified as hyperoside, icariin, epimedin B, epimedin C, inositol.

CONCLUSIONCompound I and III - V were isolated from the plant for the first time.

Epimedium ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analogs & derivatives ; chemistry ; isolation & purification

An extremely thermoacidophilic isolate K4-1 was obtained from an acidic hot spring in Teng- chong Rehai, Yunnan province. Morphology, growth characteristics, utilization of carbon compounds, en- ergy sources and 16S rRNA gene sequence of K4-1 were studied. Cells of K4-1 are irregular cocci with monotrichous flagella. The strain grew aerobically in either a lithotrophic or a heterotrophic mode. Growth on elemental sulfur occurred through oxidation of sulfur. It grew optimally at 75?C and pH 3.5. On the basis of 16S rRNA gene sequence similarity, strain K4-1 was shown to belong to genus Sulfolobus, being related to the type strains of genus Sulfolobus (86.6%~94.3% similarity), and being most closely related to strain Sulfolobus tengchongensis RT8-4 (98.9% similarity). The GenBank accession number of strain K4-1 16S rRNA gene sequence is EU729124.

OBJECTIVETo analyze the mutations in Cx30 gene in a Chinese family with hidrotic ectodermal dysplasia (HED) and to make prenatal diagnosis on the embryo which has been pregnant for 5 months.

METHODSA family including 2 affected and 4 unaffected individuals was collected, and their informed consents were obtained. The affected woman had a five-month pregnancy. An 884 bp fragment containing the whole GJB6 coding sequence was amplified by PCR and the products were bi-direction sequenced directly. The mutation was further confirmed with restriction endoenzyme digesting. On the base of successful gene diagnosis, the following detection procedure on the pregnant baby was performed. First the whole coding region of Cx30 was amplified using primers Cx30-F and Cx30-R and the PCR products were digested by Hae II. Then the PCR products were cloned into pUCm-T vector. Blue-white blot screening method and PCR-restriction endoenzyme digesting technique were used to identify the correct clones. The mutant allele clone was sequenced to confirmed the mutation.

RESULTSA heterozygous missense mutation 263C --> T in the Cx30 gene was detected in the affected little girl and her affected mother, which led to an amino acid substitution (A88V) in the second transmembrane domain of GJB6. The mutation was confirmed by Hae II digestion. A88V mutant allele cannot be cut while the wild normal allele can be cut into two fragments, 520 and 278 bp. The result of analyse on the five-month pregnancy show the embryo carried the A88V mutation too. So the embryo will be a patient.

CONCLUSIONAn A88V missense mutation in the Cx30 gene can also cause HED in Chinese Han population. Based on the gene diagnosis, prenatal diagnosis can be played using bi-direction sequencing and confirmed with restriction endoenzyme digesting.

Adult ; Amino Acid Substitution ; Asian Continental Ancestry Group ; genetics ; China ; Connexin 30 ; Connexins ; genetics ; Ectodermal Dysplasia ; ethnology ; genetics ; Female ; Fetal Diseases ; diagnosis ; ethnology ; genetics ; Genetic Testing ; Heterozygote ; Humans ; Mutation, Missense ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; methods ; Sequence Analysis, DNA

OBJECTIVETo separate the low abundance protein and establish the 2-DE synthetic map of total protein of human normal spermatozoa by using the 2-DE technology.

METHODSAll the needed human spermatozoa were collected and mixed, and proteins were extracted at one time with the method of urea/thiourea and ultra-sound. 0.8 mg, 0.6 mg, 0.5 mg, 0.3 mg sperm protein extracts were separated with 2-DE. Analyzed with MALDI-TOF-MS, PI and MW of 2 spots were obtained. Then set the 2 spots as the referent spots, different maps were compared and analyzed. At last, a synthetic map enriched with low abundance protein was obtained.

RESULTS1,080 +/- 23 protein spots have been separated on the 2-DE map with standard 0.5 mg loading amount and a synthetic map A was constructed which consist of 889 matched protein spots on the all maps with 0.5 mg loading amount. 381, 50 and 32 new spots were detected individually on the maps with 0.8 mg, 0.6 mg and 0.3 mg protein loading amount. A synthetic map with 1,352 protein spots was obtained.

CONCLUSIONLow abundance protein was separated and a synthetic map enriched with low abundant protein was obtained by changing the protein loading amount.

Electrophoresis, Gel, Two-Dimensional ; Humans ; Male ; Molecular Weight ; Proteins ; analysis ; isolation & purification ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Spermatozoa ; chemistry

Action Potentials ; Animals ; Electromyography ; Environmental Exposure ; Hypothermia ; Rats ; Rats, Wistar ; Seawater ; Thigh ; physiology ; physiopathology

AIMTo establish chromatographic fingerprint of Carthamus tinctorius L. by RP-HPLC in order to control the quality of Carthamus tinctorius L.

METHODSThe gradient elution mode was applied in chromatographic separation, and data were analysed by "Computer Aided Similarity Evaluation" software to compare the quality of Carthamus tinctorius L. samples from different habitats.

RESULTSSamples from different habitats were of high similarity, though a few samples showed evident difference in fingerprint graphics.

CONCLUSIONThe RP-HPLC fingerprint method is repeatable, feasible in analysis of Carthamus tinctorius L. and can be used in quality assessment of Carthamus tinctorius L. Chemical components in Carthamus tinctorius L. samples from various habitats are similar, and their ratios between each other are stable.

Carthamus tinctorius ; chemistry ; Chalcone ; analogs & derivatives ; chemistry ; isolation & purification ; China ; Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Flowers ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Quinones ; chemistry ; isolation & purification