According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, a full-length cDNA sequence of SQS2 from S. miltiorrhiza f. alba was cloned by the method of reverse transcription polymerase chain reaction (RT-PCR). The SmSQS2 cDNA sequence was obtained, this sequence is named SmSQS2 and its GenBank registration number is KM244731. The full length of SmSQS2 cDNA was 1245 bp, encoding 414 amino acids including 5'UTR 115 bp and 3'UTR 237 bp. Sequence alignment and phylogenetic analysis demonstrated that SmSQS2 had relative close relationship to the SQS2 of S. miltiorrhiza. The induction of E. coli [pET28-SQS2] in different temperature, induction time, IPTG concentrations and density of inducing host bacterium (A600) were performed, Shaking the culture at 30 degrees C until the A600 is approximately 0.6 and add IPTG to final concentration of 0.2 mmol x L(-1), and then the optimal expression of SmSQS2 recombinant protein were accumulated after the induction time of 20 h. The research provided important base for the study of sterol and terpene biosynthesis of SQS2 in S. miltiorrhiza f. alba.
Cloning, Molecular ; Farnesyl-Diphosphate Farnesyltransferase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; classification ; enzymology ; genetics ; Sequence Alignment

Acute Disease ; Adolescent ; Adult ; Animals ; Child ; Child, Preschool ; Female ; Foodborne Diseases ; diagnosis ; etiology ; therapy ; Humans ; Male ; Meat ; poisoning ; Middle Aged ; Pesticides ; poisoning ; Swine ; Treatment Outcome

Objective To compare four methods of monitoring vascularization of tissue engineered bone in the rhesus so as to find our the best. Methods Twenty-five lower limbs of 13 rhesuses were used in this study to make models of tibial diaphyseal defect of 20mm which were to be fixed with an AO reconstruction plate of 7 holes. The monkeys were randomly divided into five groups according to defect filling materials: group A:?-tricalcium phosphate (?-TCP) and bone marrow stromal cells (BMSCs) and blood vessel bundles; group B:?-TCP and blood vessel bundles; group C:?-TCP and BMSCs; group D:?-TCP; group E: blank. Perfusion weighted MR imaging (PWMR), X-ray, radionuclide imaging and histological examinations were carried out at weeks 4, 8, 12 postop- eratively. The maximum slope rates of the single intensity-time curve (SS_(max)) and values of baseline (Sl_(?))were calculated at the same time points. Transmittances of the X-ray films were assessed. Ratios between isotope counts in region of interest (ROI) were calculated. Chinese ink perfusion and calculation of blood vessel areas were done for histological examinations, Results Compared with other groups, the SS_(max) in group A was the highest at weeks 4, 8, 12 postoperatively. In group A, the SS_(max) at week eight was significantly higher than that at week four (P= 0. 003), and the SS_(max) and transmittance of X-ray were negatively related at week 12 after operation (rs=-0. 892, P=0. 042), but the SS_(max) and blood vessel area were positively related (rs=0. 894, P=0.041)Conclusions PWMR can be a sensitive, quantitative, noninvasive and non-radiant method to monitor vascularization of tissue engineered bone, because SS_(max) of the single intensity-time curve of PWMR can reflect the most accurately the process of vascularization of tissue engineered bone.

To simplify the macrocyclic fragment and to modify the zinc binding group of the natural product apicidin, two series of S-hexyl (heptyl) ethanethioate derivatives were designed and synthesized. Twenty-six compounds were synthesized and confirmed with 1H NMR, IR, MS and HR-MS spectrum, which were not reported. Take vorinostat as control, their antiporliferative activities against cancer cell lines, MCF-7 and HL-60, were tested with MTT assay or trypan blue staining method. Generally in both series it was found that, the chiral carbon atom at 7 position is not necessary, compounds II-1, II-3, II-6 and II-13 showed good activity on HL-60 cells in vitro, with the IC50 values less than 10 micromol x L(-1). II-7 and II-8 showed stronger activity against MCF-7 than Vorinostat, with the IC50 of 3.19 and 6.29 micromol x L(-1), respectively.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Proliferation ; drug effects ; Drug Screening Assays, Antitumor ; HL-60 Cells ; Histone Deacetylase Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Inhibitory Concentration 50 ; MCF-7 Cells ; Peptides, Cyclic ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, with the method of reverse transcription polymerase chain reaction (RT-PCR), this study cloned full-length cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase gene from Salvia miltiorrhiza bge.f.alba, this sequence is named as SmHDS and its GenBank registration number is KJ746807. SmHDS, 2 529 bp long, contains an ORF of 2 229 bp, encodes 742 amino acids, including 5' UTR 170 bp and 3' UTR 130 bp. Using bioinformatics software, having made a homology analysis of the obtained sequence, we can have a conclusion that SmHDS have a close genetic relationship with HDS of Salvia miltiorrhiza. Analysis result of prokaryotic expression revealed that in Escherichia coli, SmHDS expressed target proteins which in size are comparable with the protein predicted. Meanwhile, the 4 factors which can influence the protein expression were optimized, the 4 factors are inducing temperature, inducing time, IPTG concentrations and density of inducing host bacterium (A600). The optimal expression conditions of SmHDS were 30 degrees C until the A600 is 0.6, and add IPTG to a final concentration of 0.2 mmol x L(-1), and the induction time of 20 h. It provides theoretical basis for the further study of the function of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase in the biosynthesis of tanshinone compounds.
Cloning, Molecular ; DNA, Complementary ; genetics ; Diterpenes, Abietane ; biosynthesis ; Enzymes ; biosynthesis ; genetics ; Escherichia coli ; metabolism ; Plant Proteins ; biosynthesis ; genetics ; Salvia miltiorrhiza ; enzymology ; genetics

A 38-year-old male patient presented with multiple papules on the scalp with itching and tingling for 3 years. Skin examination showed multiple millet- to soybean-sized red papules on the scalp. Histopathological examination showed epidermal hyperkeratosis, mildly thickened prickle cell layer, proliferation of a large number of capillaries in the dermis, and perivascular infiltration of closely distributed lymphocytes and scattered eosinophils. The blood vessels were lined by hyperplastic and hypertrophic endothelial cells, some of which projected into the vascular cavity. The patient was diagnosed with angiolymphoid hyperplasia with eosinophilia. After 1 session of Nd:YAG laser therapy, all the skin lesions were removed, and no obvious adverse reactions were observed. No relapse occurred during 5 months of follow up.

The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.
Cell Separation ; methods ; Culture Media ; Pinellia ; physiology ; Protoplasts ; physiology ; Regeneration

BACKGROUNDPlatelet-rich plasma (PRP) as a storage vehicle of growth factors has been successfully used in clinical applications, but in most cases the platelets were autologous. However, the large volume of blood withdrawn has detrimental effects on patients with anemia or poor general health. To overcome these limitations, this study was designed to separate the growth factors in homologous platelet-rich plasma.

METHODSThe gel chromatography with Superdex-75 column was applied to separate PRP supernatants into 4 major fractions. Then the four fractions were vacuumed freeze-dried and re-dissolved in phosphate buffered saline. Proteins concentrations in PRP and in four fractions were detected by bicinchoninic acid protein assay; platelet derived growth factor-AB (PDGF-AB) and transforming growth factor beta1 (TGF-beta1) levels were determined by sandwich enzyme-linked immunosorbent assays. The effects of fractions on the proliferation of human marrow-derived mesenchymal stem cells (MSCs) were determined by 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay.

RESULTSPRP supernatants were separated into four major fractions by gel chromatography. The proteins recovery was 96.72%. Of the four fractions, fraction B contained the highest TGF-beta1 and PDGF-AB levels, and the highest proteins concentrations. Cell proliferation curves of MSC demonstrated that fraction B and C induced a remarkable increase of MTT values compared to the untreated culture (P < 0.05), and the effects of fraction B and C showed no significant difference compared to the PRP group (P > 0.05). Fraction A and D showed no significant difference to the negative control group (P > 0.05).

CONCLUSIONSThe growth factors in PRP supernatants could be preliminarily separated into four fractions by gel chromatography, and the freeze-drying fractions retained the biological activity of growth factors. The growth factors were mostly presented in fraction B and C, and they promoted cell proliferation effectively.

Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Chromatography, Gel ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Platelet Count ; Platelet-Derived Growth Factor ; isolation & purification ; pharmacology ; Platelet-Rich Plasma ; chemistry ; Transforming Growth Factor beta1 ; isolation & purification ; pharmacology

OBJECTIVETo explore the characters of lung injury induced by tin dusts and to provide the diagnosis evidence of tin pneumoconiosis.

METHODSForty SD rats were randomly divided into four groups: the group exposed to tin dusts from smelting workshop, the group exposed to tin dusts from tin refining workshop, the positive control group exposed to standard quartz dusts and the negative control group exposed to saline. The pathological changes of rat lungs were observed dynamically.

RESULTSIn rats exposed to tin dusts, on the 30th day after exposure to tin dusts, the scattered hoar tip size of the spots in surface and section of the lungs were observed, the scattered focal granulomatous inflammation around the small bronchi and dust particles in lung tissue were observed under microscope; on the 90th day after exposure to tin dusts, the granulomatous inflammation increase, the fibroblasts proliferation, collagen fibers formation and positive VG staining were found. There were significant differences, as compared with positive or negative controls (P < 0.05). These pathological changes were basically the characters of specific pathological changes in early tin pneumoconiosis.

CONCLUSIONNon-ferrous metal tin dusts can induce the specific lung injury (granuloma formation) in lung tissue of rats exposed to tin dusts, which fulfilled the diagnostic criteria of specific pathological changes in early tin pneumoconiosis.

Animals ; Dust ; Lung ; pathology ; Lung Injury ; chemically induced ; diagnosis ; pathology ; Rats ; Rats, Sprague-Dawley ; Tin ; adverse effects

OBJECTIVETo investigate the effect of National Natural Science Foundation of China (NSFC) on the progress of dental research from 1986 to 2010.

METHODSThe data regarding the NSFC allocated to dental and craniofacial research from 1986 to 2010 were collected. Total expenses and numbers of the majority of programs and the situation of completed program finished in recent 7 years were provided.

RESULTSFrom 1986 to 2010, a total of 922 projects and 204 401 thousands Chinese Yuan supported by NSFC were allocated to dental research. The detailed allocations were as follows: general program (564), young scientists fund (258), regional fund (40), key program (11), national science fund for distinguished young scholars (5), major international (regional) joint research program (1), others (43). The grants of talent training increased dramatically. Taking the projects (307) completed between 2003 and 2009 for example, 307 papers were published in Science Citation Index (SCI) included journals and 1049 papers were published on Chinese journals. By the time of completion of the projects, 39 post-doctoral students, 590 students for PhD degree and 670 students for Master degree had been trained.

CONCLUSIONSOver the past 25 years, the continuous increase of NSF on dental research has led to substantial achievement, resulting in great progress of dental oral-cranio-facial research.

China ; Economics, Dental ; Financial Support ; Financing, Organized ; Foundations ; economics ; Oral Medicine ; economics ; Research Support as Topic ; economics ; Retrospective Studies