A strain NK13 was screened for a certain extent asymmetric hydrolyze the rac-ketoprofen chloroethyl ester and identified as Bacillus megaterium. For the preparation of gene libraries, a positive clones was obtained from the tributyrin flat. The sequence of this esterase gene had been analysised, and contained the whole ORF of an esterase gene with the length of 933bp. The esterase gene of NK13 was compared with the esterase genes of GenBank and the result showed that the esterase gene of NK13 was a novel gene(GenBank accession nember DQ196347).The new esterase gene was inserted into the plasmid pET21b+, then the recombinant plasmid transformed E.coli BL21. After being induced by IPTG, it was expressed in the host strain. SDS-PAGE analysis showed that the relative molecular mass of the esterase was about 34kDa. The result of TLC and HPLC showed that the recombinant strain had higher conversion ration than templet strain. 47.4%of the rac-ketoprofen Chloroethl ester was hydrolyzed to ketoprofen by the recombinant strain in 45min. The (S)- ketoprofen enantiomeric excess, in the later,was 55.46%, which indicated that the esterase could hydrolyze (S)-ketoprofen chloroethyl ester firstly.

OBJECTIVETo compare the effect between mini-traumatic bone-grafting and non-bone-grafting in percutaneous K-wire fixation for treating the calcaneal fractures.

METHODSFrom 2002 to 2006, 112 patients with the type II (Paley type) fractures of calcaneus were studied. There were 56 cases in bone-grafting group involving 36 males and 20 famales,aged from 21 to 65, averaged (42.0 +/- 2.3) years; 11 cases were in type II a and 45 were in type II b; the course was from 3 to 14 days, averaged (6.0 +/- 1.2) days. And there were 56 cases in non-bone-grafting group involving 38 males and 18 famales,aged from 22 to 67, averaged (43.0 +/- 2.5)years; 13 cases were in type II a and 43 were in type II b; the course was from 2 to 15 days, averaged (5.0-2.1) days. All the cases were treated by closed reduction and percutaneous K-wire fixation, and bone-grafting group(56 cases) were treated by mini-traumatic bone-grafting, but the other group (56 cases) were not. The collapsing rate and fineness rate were compared.

RESULTSAll the cases were followed up from 5 to 52 months. There were no collapsing cases in the bone-grafting group after operation, but 3 cases occurrenced re-collapsing in the non-bone-grafting group. According to the Zhang Tie-liang's evaluation criterion, in the bone-grafting group,the results were excellent in 43 cases, good in 12, fair in 1, the fineness rate was 98.2%. In the non-bone-grafting group,the results were excellent in 37 cases, good in 16, fair in 2, poor in 1, the fineness rate was 94.7%.

CONCLUSIONTreatment of the type II fracture of calcaneus with closed reduction, percutaneous K-wire fixation and mini-traumatic bone-grafting can prevent the posterior talar articular surface of caltaneus from collapsing again after operation, enhance the union of fracture, elevate the curative effect, thus it should be taken with the standard therapeutic regimen.

Adult ; Aged ; Bone Transplantation ; methods ; Bone Wires ; Calcaneus ; injuries ; surgery ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Transplantation, Autologous ; Treatment Outcome ; Young Adult

OBJECTIVETo establish a qualitative and quantitative reversed-phase high-performance liquid chromatography (RP-HPLC) with fingerprinting technique for quality control of compound dandelion enema.

METHODSHPLC was utilized for quality assessment of 10 batches of samples. RP-HPLC analysis was performed on a Hypersil BDS C18 column (4.6 mm x 250 mm, 5 microm) with the mixture of acetonitrile (A) and potassium phosphate solution (B) (pH3.2) as the mobile phase in gradient mode. The concentrations of solvent A were 10%, 80% and 80% at 0, 38 and 40 min, respectively. The column temperature was set at 35 degrees C, the flow rate at 0.7 ml/min and the detection wavelength at 254 nm.

RESULTSHPLC fingerprinting was established from the 10 batches, and the data showed 23 characteristic peaks in the compound dandelion enema for use as index peaks for qualitative identification. Comparison of the retention time and the on-line UV spectra of the samples with the chemical standards identified peaks 3, 4 and 8 as protocatechualdehyde, caffeic acid and ferulic acid, respectively. The contents of caffeic acid in the compound dandelion enema ranged between 63.7 and 136.8 microg/ml.

CONCLUSIONHigh specific chromatographic fingerprinting and quantitative measurement of caffeic acid allows rigorous quality control of compound dandelion enema.

Caffeic Acids ; analysis ; standards ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Reference Standards ; Reproducibility of Results ; Taraxacum ; chemistry

OBJECTIVETo explore the relationship between long (L) type PML-RAR alpha fusion gene and the prognosis of patients with acute promyelocytic leukemia (APL).

METHODSPML-RAR alpha fusion gene was detected by RT-PCR in 33 APL patients. The optical density of three bands including E5 (+) E6 (+) 636 bp (no deletion), E5 (-) E6 (+) 492 bp (exon 5 deleted) and E5 (-) E6 (-) 232 bp (both exon 5 and exon 6 deleted) was measured by a UVP analysis system and their relative proportions were calculated. The relative expression level of each splicing band, initial WBC count and age were statistically (for single and multi-factor analysis) analyzed with prognosis.

RESULTSThe relative expressions of E5 (-) E6 (+) and E5 (-) E6 (-) in death group were obviously different from that in first complete remission (CR1) group (P < 0.01), but do not for E5 (+) E6 (+) (P > 0.05). The relative expression levels of E5 (-) E6 (+), E5 (-) E6 (-) and E5 (+) E6 (+) were 0.23 +/- 0.12, 0.58 +/- 0.18, 0.20 +/- 0.09 in death group, and 0.45 +/- 0.16, 0.23 +/- 0.12, 0.31 +/- 0.16 in CR1 group, respectively. Initial WBC count and age was no difference between the two groups (P > 0.05). Logistic regression analysis showed that the expression of E5 (-) E6 (+) had no effect on the prognosis (B = 3.475, P = 0.492), but the expression of E5 (-) E6 (-) showed a negative correlation with prognosis (B = -19.660, P = 0.046).

CONCLUSIONSThe high expression of E5 (-) E6 (-) is correlated with the poor prognosis for patients with APL.

Adolescent ; Adult ; Alternative Splicing ; Female ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Prognosis ; Young Adult

Most plants can form a symbiosis in root with microorganisms for mutual benefit, Nonlegumes mainly form the symbiotic mycorrhiza with arbuscular fungi. The interaction is initiated by invasion of arbuscular mycorrhizal (AM) fungi into the plant root, and follows by production of several special signal molecules, such as the symbiosis receptor-like kinase (SYMRK) from plant. SYMRK has an extracellular domain comprising three leucine-rich repeats (LRRs), a transmembrane domain and an cytoplasmic protein kinase domain. Symrk is required for a symbiotic signal transduction pathway from the perception of microbial signal molecules to the rapid symbiosis-related gene activation. Study of symrk may set up a solid foundation for giving further insight on the function and mechanism of plant-fungi symbiosis.
Amino Acid Sequence ; Host-Pathogen Interactions ; Lycopersicon esculentum ; Molecular Sequence Data ; Mycorrhizae ; physiology ; Phosphotransferases ; classification ; genetics ; Phylogeny ; Plant Proteins ; classification ; genetics ; Plant Roots ; enzymology ; genetics ; microbiology ; Sequence Homology, Amino Acid ; Signal Transduction ; genetics ; Symbiosis ; genetics

Objective To observe the brain activation of interictal epiletiform discharges(IEDs) and to localize the epileptogenic foci of epilepsy.Methods The electroencephalogram(EEG)and functional MRI data of 12 focal epileptic patients were acquired using a combination of EEG and functional MRI simultaneously.The IEDs onset time detected with EEG were set as the time parameters in an event- related paradigm of functional MRI analysis.The spatial and temporal characters of IEDs activation were analyzed in detail.In order to confirm the consistency of this method,all patients were scanned repeatedly and the results were correlated with clinical evaluation.Results Of the 12 patients,valid data from EEG- fMRI were obtained from 10 patients in a total of 18 sessions..Compared with the structural foci,the epileptic foci localization results of eleven sessions were good,five sessions were fairly good,and two sessions were poor.The results obtained from six patients in two separate sessions were concordant.respectively.Moreover,thalamic activation was detected in ten sessions,cerebellar activation was detected in all sessions,and the deactivation was found in the default mode loci in nine sessions. Conclusion The method of performing EEG and fMRI simultaneously can potentially be a useful tool in epilepsy research.

Adult ; CD5 Antigens ; metabolism ; Carcinoma, Papillary ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Choristoma ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Hamartoma ; metabolism ; pathology ; Humans ; Proto-Oncogene Proteins c-kit ; metabolism ; Thymoma ; metabolism ; pathology ; Thymus Gland ; pathology ; Thymus Neoplasms ; metabolism ; pathology ; Thyroid Neoplasms ; metabolism ; pathology ; surgery ; Thyroidectomy ; methods

Osteosarcoma is suggested to be caused by genetic and molecular alterations that disrupt osteoblast differentiation. Recent studies have reported that transmembrane protein 119 (TMEM119) contributes to osteoblast differentiation and bone development. However, the level of TMEM119 expression and its roles in osteosarcoma have not yet been elucidated. In the present study, TMEM119 mRNA and protein expression was found to be up-regulated in osteosarcoma compared with normal bone cyst tissues. The level of TMEM119 protein expression was strongly associated with tumor size, clinical stage, distant metastasis and overall survival time. Moreover, gene set enrichment analysis (GSEA) of the Gene Expression Omnibus (GEO) GSE42352 dataset revealed TMEM119 expression in osteosarcoma tissues to be positively correlated with cell cycle, apoptosis, metastasis and TGF-β signaling. We then knocked down TMEM119 expression in U2OS and MG63 cells using small interfering RNA, which revealed that downregulation of TMEM119 could inhibit the proliferation of osteosarcoma cells by inducing cell cycle arrest in G0/G1 phase and apoptosis. We also found that TMEM119 knockdown significantly inhibited cell migration and invasion, and decreased the expression of TGF-β pathway-related factors (BMP2, BMP7 and TGF-β). TGF-β application rescued the inhibitory effects of TMEM119 knockdown on osteosarcoma cell migration and invasion. Further in vitro experiments with a TGF-β inhibitor (SB431542) or BMP inhibitor (dorsomorphin) suggested that TMEM119 significantly promotes cell migration and invasion, partly through TGF-β/BMP signaling. In conclusion, our data support the notion that TMEM119 contributes to the proliferation, migration and invasion of osteosarcoma cells, and functions as an oncogene in osteosarcoma.
Apoptosis ; Bone Cysts ; Bone Development ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Movement ; Dataset ; Down-Regulation ; Gene Expression ; In Vitro Techniques ; Neoplasm Metastasis ; Oncogenes ; Osteoblasts ; Osteosarcoma* ; RNA, Messenger ; RNA, Small Interfering ; Up-Regulation*