To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.

Acyl carrier protein is an essential component involved in the biosynthesis of DHA(Docosahexaenoic Acid) via PKS(Polyketide synthase) pathway,which takes the growing acyl chain from one enzyme to another.One cDNA clone,with high homology of ACP,was isolated from Schizochytrium sp.FJU-512 cDNA library.The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4'-phosphopantetheine prosthetic(4'-PP) binding site.The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp.The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis demonstrated that ACP was effectively expressed.

The method of triggering the respirator to perform airfeed by the management of the diaphragmatic electromyography (EMG) signal is introduced in this paper. By using LabVIEW, an analysis software of the diaphragmatic electromyographic (EMG) signals is developed to filter the diaphragmatic EMG signal. The characteristics of the filtered signal is analyzed to catch the inspiratory onset, and then a series of signals are sent out duly to trigger the respirator to perform airfeed, and thus the patient's respiration can be synchronized with the respirator.

OBJECTIVETo study the effect of Kanglaite injection on cyclooxygenase activity in lung carcinoma A549 cells.

METHODSThe expression of mRNA of COX-1 and COX-2 were measured by RT-PCR. The expression of COX-2 protein was measured by Western-blot.

RESULTKanglaite injection could selectively decreased COX-2 mRNA expression and protein expression while COX-1 mRNA expression was unchanged.

CONCLUSIONKanglaite injection is selectively inhibitory agent of COX-2, and possess inhibitory effects on cyclooxygenase 2.

Antineoplastic Agents, Phytogenic ; administration & dosage ; isolation & purification ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; enzymology ; pathology ; Cell Line, Tumor ; Coix ; chemistry ; Cyclooxygenase 1 ; biosynthesis ; genetics ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; Injections ; Lung Neoplasms ; enzymology ; pathology ; Plant Oils ; administration & dosage ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Seeds ; chemistry

Objective To study the CT and MRI findings of osteosarcoma in paranasal sinus and evaluate their clinical value.Methods All 9 cases of osteosarcoma were verified by histopathology.Imaging data were analyzed retrospectively.Results The lesion occurred in maxillary sinus in 5 cases,in ethmoid sinus in 3 cases and in sphenoid sinus in one case.Primary osteosarcoma was found in 7 cases.Secondary osteosarcoma occurred from fibrous dysplasia and ossifying fibroma each in one case.On CT,the involved areas revealed bony destruction associated with ill-defined and irregular soft tissue mass.The lesion showed heterogeneous density with minimal or massive tumor bone formation,cloud-like in 3 cases,ivory-like in 2 cases,spicule-like in 2 cases,cloud- and spicule -like in one case and spicule- and ivory-like in one case.Postcontrast CT showed mild to moderate inhomogeneous enhancement in 3 cases.On MR T_1 WI,the lesions showed hypointensity compared to brain in 5 cases and iso-intensity in 2 cases.On T_2WI,the lesions showed heterogeneous hyperintensity in 4 cases and isointensity in 3 cases with marked hypointense foci which corresponded to tumor bone on CT.Postcontrast MR imaging demonstrated moderate to marked inhomogeneous enhancement in these cases.MRI showed accurately the extent and associated changes of the lesions.The lesions invaded the orbit,pterygopalatine and infratemporal fossae,skull base and extensive craniofacial structures in 5,4,3 cases and 1 case,respectively.Conclusion CT is the optimal modality in showing tumor bone of osteosareoma in paranasal sinus.MRI can demonstrate optimally the invading extent of the lesions.Combined imaging modalities can provide more comprehensive information for diagnosis and therapy of osteosarcoma in paranasal sinus.

Objective To evaluate the feasibility of 3-(4-~(18)F-fluorobenzyl)-8,9-dimethoxy-1,2,3,4-tetrahydrochromeno [3,4-c]pyridin-5-one ( is F-FDTP) as a potential dopamine D4 receptor PET imaging agent.Methods ~(18)F-FDTP solution in ethanol-physiological saline was incubated with calf serum to test its in vitro stability through the determination of radiochemical purity.Normal rats were injected intravenously with ~(18)F-FDTP and then sacrificed at 2,5,10,15,30,60 and 120 min after anesthesia.Blood,organs and brain tissue samples were collected.All samples were weighed and measured for radioactivity.The uptake of samples was expressed as percentage activity of injection dose per gram of tissue ( % ID/g).Results The stability of ~(18)F-FDTP was satisfactory and its radiochemical purity was above 95% after incubation 120 min at 37℃ in calf serum.The biodistribution showed that ~(18)F-FDTP could penetrate through the blood-brain barrier and selectively accumulate in striatum,hypothalamus,frontal certex,hippocampus,cerebellum,where the D_4 receptor was reportedly located.The radioactivities in hippocampus,hypothalamus,striatum,frontal cortex,cerebellum,pons were (0.42±0.03),(0.46±0.05),(0.54±0.04),(0.39±0.04),(0.45±0.06),(0.35±0.04) %ID/g,respectively,2 min post injection.And there was difference between the normal biodistribution results and the blocking experimental results:(0.36 ±0.05),( 0.33±0.05 ),(0.55±0.05 ),(0.30±0.07 ),(0.34±0.07 ) and (0.32±0.04) % ID/g in hippocampus,hypothalamus,striatum,frontal cortex,cerebellum and pons,respectively.Conclusions ~(18)F-FDTP can penetrate through the blood-brain barrier and selectively accumulate in striatum,hypothalamus,frontal cortex,hippocampus,cerebellum,where the D_4 receptor was known to concentrate.These preliminary results suggest that ~(18)F-FDTP is a potential dopamine D_4 receptor imaging agent and further studies are needed.

OBJECTIVETo observe the clinical efficacy of Tiaozhi Jiangtang Tablet (TZJT) on patients suffering from diabetes mellitus with blood stasis syndrome.

METHODSSixty patients were randomly assigned into 2 groups, the TZJT group (n = 30) treated with TZJT tablet and asprin, the control group (n = 30) treated with asprin alone.

RESULTSThe improvement of symptoms was more significant in the TZJT group than that in the control group (P < 0.05). Levels of serum endothelin (ET), nitric oxide (NO) content and blood viscosity were decreased in both groups after treatment, and the effect of TZJT plus asprin was superior to that of asprin alone.

CONCLUSIONTZJT combined with asprin is effective in improving the serum content of ET and NO and reducing blood viscosity.

Adult ; Aged ; Aged, 80 and over ; Coronary Artery Disease ; drug therapy ; etiology ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy

OBJECTIVETo establish human colorectal crypt cell line.

METHODSColorectal crypt cells were separated from human fetal gut by dispase I digestion, AKP-negative cells from fetal colorectal crypt were collected and cultured on Matrigel matrix. Subsequently the primary cultured cells were transfected with recombinant retrovirus containing human telomerase reverse transcriptase (hTERT) and simian virus 40 large T antigen (SV40 LT) in 48 h. The characterization of immortalized cells was identified after the transfection and cells were screened with antibiotics for 12 approximately 16 weeks and expanded.

RESULTSMucin, cytokeratin-pan, 8, 19 were presented in immortalized cells by immunohistochemical staining; ectopic expressions of both hTERT and SV40 LT were also found in immortalized cells by Western blotting. Agarose electrophoresis showed that the cells expressed Musashi-1 mRNA. No evidence of carcinogenesis was found in nude mouse experiment and soft-agarose cloning test.

CONCLUSIONThe immortalized human colorectal crypt cells were characterized and the established cell line may be an ideal target for carcinogenesis study in vitro.

Cell Line, Transformed ; Colon ; cytology ; DNA-Binding Proteins ; Fetus ; Humans ; RNA, Neoplasm ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Simian virus 40 ; immunology ; Stem Cells ; cytology ; Telomerase ; genetics ; metabolism ; Transcription, Genetic ; Transfection ; methods

Animals ; Bone Morphogenetic Protein Receptors ; Bone Morphogenetic Proteins ; genetics ; physiology ; Cell Differentiation ; Gene Expression Regulation ; Hair Follicle ; cytology ; embryology ; physiology ; Humans ; Receptors, Growth Factor ; analysis ; Stem Cells ; cytology

OBJECTIVETo determine whether testosterone-induced intra-testicular testosterone withdrawal and therefore spermatogenic impairment is associated with looser arrangement of spermatogenic cells in rats.

METHODSAdult male SD rats received intramuscular injection of testosterone undecanoate at 19 mg/(kg x 15 d) for 130 days, and then testicular tissue blocks were obtained for the preparation of methacrylate resin-embedded sections and observation of the changes in testicular histology.

RESULTSApart from such changes as impaired spermiogenesis and spermiation, apparently looser arrangement of spermatogenic cells was seen in 11.5% of the seminiferous tubule profiles, with radial cracks (empty spaces) running towards the tubule lumen being formed between lines, bundles or groups of spermatogenic cells (mainly spermatids and spermatocytes).

CONCLUSIONLooser arrangement of spermatogenic cells is one of the key histological changes resulting from intra-testicular testosterone withdrawal in rats.

Animals ; Male ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; cytology ; drug effects ; Spermatogenesis ; drug effects ; Testis ; cytology ; drug effects ; pathology ; Testosterone ; administration & dosage ; adverse effects