Objective To investigate the effect and mechanism of dexmedetomidine, mifepristone and dexmedetomidine plus mifepristone on the fear memory in rats with post?traumatic stress disorder ( PTSD) . Methods 40 male SD rats were randomly divided into five groups with 8 in each group:control group (group C),PTSD model group (group P),dexmedetomidine group (group D),mifepristone group ( group M) and dexmedetomidine plus mifepristone group ( group U) . Fear memory in rats was evaluated by fear conditioning test ( FC) . Anxiety?like behavior was assessed by the elevated plus?maze test ( EPM) . Ex?pressions of BDNF and its receptor TrkB in the hippocampus of rats after fear condition were detected using Western blot ( WB) and CORT level in the serum was detected using enzyme?linked immunosorbent assay (ELISA). Results Compared with group P,the freezing scores in the FC in group D((32.29±8.09) %), M((33.33±8.21) %),and U((9.38±3.31) %) were significantly decreased (P<0.05). The times and en?tries in the open arms of the EPM were significantly increased (P<0.05) . The expressions of BDNF in group D(0.65±0.04),M(0.71±0.04),U(0.79±0.07) and TrkB in group D(0.66±0.04),M(0.71±0.04),U (0.86±0.03) were obviously rescued in hippocampus of rats (P<0.05). The CORT level in serum in group D ((37.65±12.37)μg/L) and U((59.10±5.23)μg/L) was decreased (P<0.05). There was no difference be?tween group P and M. Conclusion These results suggest that dexmedetomidine, mifepristone and dexme?detomidine plus mifepristone can significantly enhance fear extinction and improve anxiety?like behaviors in rats with PTSD. The mechanism may be that dexmedetomidine and mifepristone could enhance the expres? sions of BDNF and TrkB in the hippocampus.

ObjectiveStromal cell-derived factor -1 (SDF-1 ) is closely related to the biological characteristics of breast cancer. We aimed to explore whether estrogen affected breast cancer by SDF-1. MethodsThe breast cancer cell line MCF-7 and MRC5 were chosen, and divided into three groups: the control group, the estrogen group and the estrogen + estrogen receptor blocker group. Each group was cultured with different physiological concentrations of 17-β estrogen at certain time, and the same alcohol concentration of 17-β estradiol at different time points, and then the enzyme-linked immunosorbent assay (ELISA) was used to measure the concentration of SDF-1 in culture medium, and the semi-quantitative reverse transcriptionpolymerase chain reaction (RT -PCR) was used to detect the expression of SDF-1 mRNA in each group.ResultsSDF-1 can be detected in the culture medium of both MCF-7 and MRC5 cell lines. All different concentrations of 17-β estradiol may increase the secretion of SDF-1 in MCF-7 cells. When adding 17-β estradiol to the concentration of 107mol/L, the secretion of SDF-1 reached the peak in 2 hours, which was 6 times and 2.7 times that of control group ( P ＜ 0.01 ). The effect could be ehminated by pure estrogen receptor ICI182,780. In addition, the mRNA expression of SDF-1 was consistent with the SDF-1 protein levels-l07 mol/L group. The expression of SDF-1 mRNA was higher than both that of the control group and the blocking group in 2 hours (P ＜ 0.05 ). ConclusionsIn some breast cancer cell lines, physiological concentrations of estrogen can increase the secretion of SDF- 1, and this effect is mainly achieved through the estrogen receptor. Estrogen can influence the biological characteristics of breast cancer by SDF-1.

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

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

The safflower floret is a traditional Chinese medicine used to promote blood circulation and remove obstruction in the channels. The spines on its bracts are considered a handicap when manual harvest is involved. In this study, cDNA-SRAP was used to systematically investigate which genes are associated with the spines. Sixty pairs of possible primer combinations were used on two cDNA pools representing spininess and spinelessness. Six transcript-derived fragments were identified, of which two with low recombination were sequenced successfully and named as GPY-1 and GPY-2. By using the RACE method, the full-length cDNA of GPY-2 is cloned and named as CTL-spn. The full-length cDNA of CTL-spn was 1 679 bp long with a 1 524 bp ORF encoding a 508 aminoacid protein. The deduced amino acid sequence of the CTL-spn gene shared a high homology (97%) with other known ATP synthase CF1 alpha subunits. Semiquantitative RT-PCR analysis revealed that the mRNA of GPY-1 and GPY-2 accumulated in only spiny lines. Considering the important role of ATP synthase CF1 alpha subunit in plants, it may directly take part in the formation process of spininess and enhancing resistance reaction of spiny safflower. Also, our results provide the important insights for breeding spineless cultivars of safflower.
Adenosine Triphosphate ; Amino Acid Sequence ; Carthamus tinctorius ; enzymology ; genetics ; Chloroplast Proton-Translocating ATPases ; genetics ; DNA Primers ; DNA, Complementary ; Plant Proteins ; genetics

Diagnosis, Differential ; Esophageal Neoplasms ; pathology ; surgery ; Granuloma, Pyogenic ; pathology ; surgery ; Hemangioma, Capillary ; pathology ; Hemangiosarcoma ; pathology ; Humans ; Male ; Middle Aged ; Sarcoma, Kaposi ; pathology

Objective To investigate the effect of genistein on Notch-1, SHH and HHIP gene expression and on the cell cycle and proliferation of of BxPC3 cells. Methods Human pancreatic cancer cell line BxPC3 was cultured. The BxPC3 cells were treated with genistein and then the total RNA and protein were extracted. RT-PCR was used to detect the expression of Notch-1 mRNA, SHH mRNA and HHIP mRNA. Noteh-1 and SHH protein was determined by western blotting. MTT assay was used to detect proliferation of BxPC3 cells. The cell cycle of BxPC3 cells was measured by Propidium iodide (PI) and flow cytometry. Results The inhibiting rate was 67.17%±2.32% when BxPC3 cell lines were treated by 20μg/ml genistein for 48 hours. Notch-1 mRNA was down-regulated from 2.454±0.068 to 1.304±O.169 ; SHH mRNA was down-regulated from 0.959±0.023 to O.472±0.077 ; HHIP mRNA was up-regulated from 0.625±O.158 to 1.761±0.121. Notch-1 protein expression was down-regulated from 1.361±0.109 to 0.760±0.114; SHH protein expression was down-regulated from 0.265±0.018 to 0.129±0.013. (52.77±9.47)% cells were hindered in G2/M stage. Conclusions Genistein could down-regulate Notch-1 expression and inactivate Hedgehog signaling pathway and inhibit the proliferation of pancreatic cancer cells.

Objective To investigate the direct inhibitory effects of 153Sm- DTPA-c (Cys-Gly-Arg-Arg-Ala-Gly-Gly-Ser-Cys) NH2 ( 153 Sm-DTPA-c (CGRRAGGSC)) on human prostate cancer PC-3 cells. Methods 153Sm-DTPA-c(CGRRAGGSC) was synthesized by the reaction of 153SmCl3 with DTPA-c(CGRRAGGSC) using indirect synthesis method. PC-3 cells in vitro culture were divided into four study groups, groug A ( the control, with PBS only), group B with 1.5 mg/L c ( CGRRAGGSC), group C with 370 kBq 153 SmCl3 and group D with 370 kBq 153Sm-DTPA-c(CGRRAGGSC). PC-3 cell growth was assayed by 3-(4, 5-dimethylthiazol-2-yl ) -2, 5-diphenyltetrazolium bromide (MTT) method. Cell cycle and apoptosis were analyzed by flow cytometry. The expression changes of interleukin 11 (IL11 ) and IL11 receptor (IL1 1 R) in PC-3 cells were examined by Western Blot. One way analysis of variance (ANOVA) and paired-t test were applied for statistic analysis. Results The labeling yield of 153Sm-DTPA-c (CGRRAGGSC) was 85% and the radiochemical purity was 95.8%. The specific activity of 153Sm-DTPA-c(CGRRAGGSC) was 1.32 × 105 MBq/μmol. Significant inhibitory effects on the growth of PC-3 cells were found in both group C and D, with a time-dependent manner. However, no obvious inhibition was found either in group A or in group B. After 48 h,significant differences of sub-G1 peak area were found among groups, (0. 98 ± 0. 18)%, (0. 35 ±0. 10)%, (4.05 ±0.28)% and (13.38 ±0. 89)% for group A, B, C and D, respectively. Furthermore,sexpression of ILl 1R in group D was significantly lower than that in group B and C with absorbance values 0. 339 ~ 0.014, 0.338 ~ 0.019, 0.226 ~ 0. 015 for group B, C and D, respectively. Absorbance values in groups B and C were not significantly different after treatment, compared with those before treatment; however, there was difference between absorbance values after and before treatment in group D ( t = 0. 405,1. 163,135.989,P>0.05 >0.05, <0.05). Conchluion 153Sm-DTPA-c(CGRRAGGSC) can directly in hibit the cell growth and expression of human prostate cancer cells PC-3.

Objective · To establish human cervical cancer xenografts in nude mice and investigate the antitumor therapeutic effects and safety of EGF modified cisplatin-alginate conjugate liposomes. Methods · Cervical cancer-bearing mouse model was established by subcutaneously injecting Hela cells in nude mice. Generalstate and xenograft growth of the mice were observed. Tumor volumes, tumor weights, and the body weights of mice during the treatment were recorded and analyzed. The expression levels of EGFR in xenografts were detected by immunohistochemistry. Results · ① EGFR was highly expressed in the xenografts. ② CS-EGF-Lip inhibited the tumor growth effectively (P=0.000). ③ The inhibition rates of tumor volume and tumor weight of CS-EGF-Lip were 80.22% and 58.60% respectively, which were betterthan those of cisplatin (P=0.000). ④ CS-EGF-Lip had minimal influence on body weight in mice (P=0.000). Conclusion · CS-EGF-Lip has more effective antitumoreffects than cisplatin in cervical cancer-bearing mice, which can inhibit tumor growth of solid tumors with enhanced efficacy and safety.

BackgroundFlap creation is one of the most important steps during laser in situ keratomileusis (LASIK). As the microkeratome blade technology is developing, the accuracy, uniformity and reproducibility of corneal flaps created by the microkeratome blade are of high clinical concern. ObjectiveThe aim of this trial was to compare the features of corneal flaps created using the Moria M2 microkeratome 110 μm-knife with regular blade versus the Med-Logics O blade. MethodsA pilot and prospective study was designed. Two hundred and four eyes of one hundred and two patients were enrolled in this clinical trial. The patients were divided into the Moria M2 microkeratome 110 μm-knife with Med-Logics 0 blade group ( 110-0 group) ( 102 eyes) and Moria M2 microkeratome with 110 μm-knife with regular blade group (110 group) (102 eyes),with the matched demography. Fourier-domain optical coherence tomography ( RTVue OCT) was used to measure flap thickness using 28 settings on the 204 corneas at one week postoperatively. The features of the LASIK flaps were analyzed on the basis of the outcomes. Written informed consent was obtained from each patient prior to LASIK. Results There was no statistically significant difference in uncorrected visual acuity and the mean spherical equivalent between the 110-0 group and 110 group ( Z =-0. 375,P =0. 708 ; u =0. 056, P =0. 956 ) one week after LASIK. The mean flap thickness of the 110-0 group was considerably thinner than that of the 110 group ( 133.28+15.41μ m versus 142.81 ±10. 07μm) ( u =-5. 227,P＜0. 001 ). The corneal flaps in both the 110-0 group and in 110 group showed a meniscus shape. The nasal flap thickness of the right eyes was not evidently different from that of temporal ( P＞0. 05 ) , but in the left eyes, nasal flap thickness was obviously thicker than the temporal flap thickness (P＜0. 05) in both groups. The mean deviation between the achieved and attempted flap thickness ( 130 μm) were (17.46±2.28) μm in the 110-0 group and ( 16. 82±6. 12) μm in the 110 group, showing a significant difference between them ( u ==0. 517, P=0. 608 ).ConclusionsThe shape of flaps created using the Moria M2 110-0 is more uniform and closer to the expected thickness of 130 μm than the ones created using the Moria M2 110 microkeratome.