Objective To investigate the effects of eritoran on the expressions of the inflammatory cytokines interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α) and interferon-[ (IFN-β) mRNA in the basilar artery after subarachnoid hemorrhage (SAH) in rabbits.Methods Atotal of 36 healthy adult male New Zealand white rabbits were randomly allocated into three groups:SAH (n =12),normal saline (n =12) and eritoran (n =12) groups.A SAH model was induced by injection of autologous arterial blood into cisterna magnatwice.An equal amount of cerebrospinal fluid was displaced with the saline in the normal saline group.An equal amount of autologous non-heparinized arterial blood was injected immediate after the replacementof cerebrospinal fluid in the SAH group.Eritoran 1.5 mg/kg was injected intravenously immediately after the blood injection via the cisterna magna each time in the eritoran group.The food intake and neurological deficit were assessed.The expressions of IL-1β,TNF-α and IFN-β mRNA in the basilar artery were detected by real-time fluorescence quantitative polymerase chain reaction.Results The food intake scores (1.20 ± 0.41 vs.2.20 ±0.61; t =53.073,P =0.002),the neurological deficit scores (1.46 ± 0.32 vs.2.6 ± 0.08; t =306.431,P =0.001),the expressions of IL-1β (1.22 ±0.48 vs.2.38 ±0.06,P =0.000),TNF-α (1.39 ±0.07 vs.3.32 ±0.21,P =0.000) and IFN-β (1.51 ±0.08 vs.2.18 ±0.05,P =0.000) in Eritoran group were all significantly lower than those in the SAH group.Conclusions Eritoran may downregnlate the expressions of inflammatory cytokines IL-1β,TNF-α and IFN-β mRNA in the basilar artery after SAH in rabbits,increasing food intake,and improving neurological deficits.

Child, Preschool ; Female ; Humans ; Infant ; Interleukin-1beta ; blood ; cerebrospinal fluid ; Male ; Seizures, Febrile ; blood ; cerebrospinal fluid ; Zinc ; blood ; cerebrospinal fluid

Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways ; Ginsenosides ; biosynthesis ; Glycosyltransferases ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Synthetic Biology

Cerebral Infarction ; blood ; Humans ; Interleukin-10 ; blood ; Interleukin-17 ; blood

This study examined the osteogenic effect of electromagnetic fields (EMF) under the simulated in vivo conditions. Rat bone marrow mesenchymal stem cells (BMSCs) and rat osteoblasts were co-cultured and exposed to 50 Hz, 1.0 mT EMF for different terms. Unexposed single-cultured BMSCs and osteoblasts were set as controls. Cell proliferation features of single-cultured BMSCs and osteoblasts were studied by using a cell counting kit (CCK-8). For the co-culture system, cells in each group were randomly chosen for alkaline phosphatase (ALP) staining on the day 7. When EMF exposure lasted for 14 days, dishes in each group were randomly chosen for total RNA extraction and von Kossa staining. The mRNA expression of osteogenic markers was detected by using real-time PCR. Our study showed that short-term EMF exposure (2 h/day) could obviously promote proliferation of BMSCs and osteoblasts, while long-term EMF (8 h/day) could promote osteogenic differentiation significantly under co-cultured conditions. Under EMF exposure, osteogenesis-related mRNA expression changed obviously in co-cultured and single-cultured cells. It was noteworthy that most osteogenic indices in osteoblasts were increased markedly after co-culture except Bmp2, which was increased gradually when cells were exposed to EMF. Compared to other indices, the expression of Bmp2 in BMSCs was increased sharply in both single-cultured and co-cultured groups when they were exposed to EMF. The mRNA expression of Bmp2 in BMSCs was approximately four times higher in 8-h EMF group than that in the unexposed group. Our results suggest that Bmp2-mediated cellular interaction induced by EMF exposure might play an important role in the osteogenic differentiation of BMSCs.

Objective To explore the value of 64-row helical CT multi-phase enhancement scan combined with angiography (CTA)in the diagnosis of pulmonary mass.Methods Two hundred and sixty-five patients with pulmonary mass confirmed by pa-thology were checked,analyzed the CT sign of multi-phase enhancement scan and the blood supply of pulmonary mass displayed by CTA.Results Lung cancer was mainly supplied by bronchial arteries,some by body arteries,the feeding arteries display rate of lung cancer group was significantly higher than that of benign disease group(P <0.05).CT enhancement peak value of lung cancer group was significantly higher than that of tuberculoma group,inflammatory pseudotumor group and hamartoma group(P <0.05), but no significant difference between hemanginoma group(P >0.05).Enhancement dynamic curves of lung cancer group was differ-ent from benign lesion groups:Lung cancer without obvious enhancement in pulmonary artery phase,CT value increased rapidly in aorta phase,120 s reached peak,and declined slowly in delay phase;CT value of tuberculoma was increased slowly without obvious peak;CT value of inflammatory increased gradually in pulmonary artery phase,90 s reached the peak;hamartoma was no obvious enhancement;Hemangioma enhanced rapidly after strengthening in the pulmonary artery phase,reached the peak at about 15 s,and then decreased slowly.Conclusion 64-row helical CT multi-phase enhancement scan combined with angiography have important clinical value,which can differentiate malignant mass from benign ones.

OBJECTIVETo explore the effect of SF-36 scale being applied in different countries under different culture and to describe the quality of life of pulmonary tuberculosis patients in China and Thailand.

METHODSSF-36 scale was applied to pulmonary tuberculosis patients in both countries using face to face interview.

RESULTSMany coefficients among domains were greater than 0.5 when quality of life of tuberculosis patient in both countries was measured. Cronbach's coefficient of all domains were greater than 0.7 for tuberculosis patients in China while cronbach's Coefficient of most domains were equal or greater than 0.7 for tuberculosis patients in Thailand except for vitality and social domains. The score of social domain for patients in Thailand was greater than that of China.

CONCLUSIONStructure validity was not good for tuberculosis patients in both countries since there were some items overlapped in different domains. However, the reliability was good for measuring quality of life of tuberculosis patients both in China and in Thailand.

China ; epidemiology ; Female ; Humans ; Male ; Quality of Life ; Reproducibility of Results ; Surveys and Questionnaires ; Thailand ; epidemiology ; Tuberculosis, Pulmonary ; physiopathology ; psychology

Lanosterol synthase is encoded by the erg7 gene and catalyzes the cyclization of 2, 3-oxidosqualene, which is a rate-limiting step of the inherent mevalonate (MVA)/ergosterol metabolic pathway in Saccharomyces cerevisiae. The intermediate 2, 3-oxidosqualene is also the precursor of triterpenoids. Therefore, the cyclization of 2, 3-oxidosqualene is the key branch point of ergosterol and triterpenoids biosynthesis. Down-regulation of 2, 3-oxidosqualene metabolic flux to ergosterol in S. cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway reconstructed by the synthetic biology approach. To construct erg7 knockout cassette harboring the loxP-Marker-loxP element, long primers were designed, which were homologous to the sequences of both erg7 ORF and plasmid pUG66. The cassette was transformed into diploid wild strain INVSc1 by LiAc/SS Carrier DNA/PEG method and then erg7 gene haploid deficient mutant was obtained by homologous recombination. The results of semiquantitative PCR and real-time quantitative PCR revealed that erg7 expression level in erg7 gene haploid deficient mutant is one time lower than that in wild strain. The results of TLC and HPLC showed that the ergosterol content in deficient mutant decreased to 42% of that in wild strain.
Chromatography, High Pressure Liquid ; DNA Primers ; Down-Regulation ; Ergosterol ; metabolism ; Haploidy ; Intramolecular Transferases ; genetics ; Polymerase Chain Reaction ; Saccharomyces cerevisiae ; genetics ; Squalene ; analogs & derivatives ; metabolism

Pichia pastoris is one of the most important systems used in the field of molecular biology for the expression of recombinant proteins. The system has advantages of high expression, high stability, high secretion, easy high-density fermentation and low cost. Many factors affect the expression of recombinant protein, such as gene copy number, codon usage preference, type of promoter, molecular chaperones, glycosylation, signal peptide and fermentation process. In this review, research advances of the above aspects are summarized, which lay a foundation for improving the expression of recombinant proteins in P. pastoris.
Fermentation ; Gene Dosage ; Glycosylation ; Molecular Chaperones ; Pichia ; metabolism ; Promoter Regions, Genetic ; Protein Sorting Signals ; Recombinant Proteins ; biosynthesis

Objective To study the in vitro effects of different doses and different kinds of LMWH on CR, and to determine whether the CR test could be used to monitor LMWH. Methods The CR value was measured with different reagents ( glass beads, celite and kaolin ) in blood samples from twenty volunteer donors, which were spiked with increasing concentration of LMWH ( dalteparin, 0-1.8 IU/ml ). Then the CR test was performed again on the same blood samples spiked with the same concentration ( 0. 8 IU/ml ) but different LMWH ( dalteparin, enoxaparin and nadroparin ). Regression analysis was performed to establish a regression equation from corresponding LMWH levels. Results With the increasing of dalteparin dose, CR values were reduced gradually for all three reagents. When the concentration of dalteparin was 0-1.8 IU/ml,the value of CR was 20. 0-4. 5 IU/min for glass beads, 26. 1-6.6 IU/min for celite and 27. 2-7. 5 IU/min for kaolin. An exponential relationship was observed between the CR value and dalteparin concentration for three reagents( R2 = -0.796, -0.884, -0.921 ,P ＜0.01 ). All three kinds of LMWH with the same concentration (0.8 IU/ml ) induced a different change in CR. The value of CR was 7.4 IU/min with dalteparin,8. 5 IU/min with enoxaparin and 8.5 IU/min with nadroparin. Compared with the control group ( CR was 17.6 IU/min ), three kinds of LMWH had statistical significance ( t = 18.45, 12. 33, 14. 93, P ＜ 0.01 ).Compared with the enoxaparin and nadroparin, dalteparin induced a higher CR value ( t = 2. 552,2. 924,P＜0. 05 ). Conclusions There is an exponential relationship between CR value and dalteparin concentration for three reagents. Three kinds of LMWH can significantly reduce the value of CR. CR test can be used to monitor the anticoagulant effect of LMWH.