Objective To construct pcDNA3-HIF-1? eukaryotic expression vector and to investigate its function in breast cancer MCF-7 cells.Methods RT-PCR was applied to amplify human HIF-1? cDNA from MCF-7 cells with a pair of sequence specific primers carrying a restriction enzyme site XbaⅠ or HindⅢ on each 5′end.HIF-1?(cDNA) was inserted into pMD18-T after sequencing,and then inserted into pcDNA3 vector.The recombinant vector was transfected into MCF-7 cells to observe its expression and function by Western blot and dual-luciferase reporter gene assay.Realtime-PCR was performed to detect the inducible expression of C-MET mRNA by HIF-1?,the anti-apoptotic effect of HIF-1 under hypoxia was analysed by detecting the Caspase3/7 activity.Results The sequence of pcDNA3-HIF-1? is identical with the gene bank.It enhances the expression of HRE reporter gene and C-MET mRNA,but decreases the Caspase3/7 activity in MCF-7 cells under hypoxia.Conclusion The pcDNA3-HIF-1? eukaryotic expression vector was successfully constructed,it not only has strong DNA binding and inducing activity,but also has anti-apoptotic effect in hypoxia MCF-7 cells.

Objective To construct pGL3-EPO-HRE reporter gene vector and to detect its function.Methods To synthesize two single-strand DNA fragments which had overlap sequence at the 3'end,then mixed them to construct the double-strand DNA fragment which include three copies of(57 bp) human erythropoietin(EPO) HRE.There was MluⅠor BglⅡ digestive site at the end of this DNA fragment.After inserting the(87 bp) fragment into pMD18-T for sequencing,the correct sequence was inserted into pGL3-promoter to construct pGL3-EPO-HRE.The recombinant vector was co-transfected into MCF-7 cells with pcDNA3-HIF-1?or pcDNA3 respectively,(12 h) later cells were treated with(100 ?mol/L) cobalt chloride(CoCl_(2)) for(24 h), then the expression of pGL3-EPO-HRE was observed by dual-luciferase reporter gene assay method.Results The sequence of pGL3-EPO-HRE was identical with human EPO HRE sequence.pGL3-EPO-HRE luciferase activity increased about 10 fold(P

BackgroundIt has been proved that,after being forced,the biological soft tissue has stable biomechanical characteristics.However,there is rare study on corneal biomechanics.Rabbit is a main animal for experimental study in ophthalmology.But the biomechanical study of cornea in normal rabbit has not been reported.ObjectiveThis study was to investigate the biomechanical properties of normal rabbit central cornea and acquire the parameter. Methods Ten rabbits were sacrificed and the whole corneas were obtained and 20 central cornea specimens with 7 mm×5 mm of rabbit were prepared and tested on BOSE electroforce 3220-AT biomechanics machine under the room temperature and suitable humidity environment.Uniaxial tension,stress between strain,relaxation and creep were performed and the curves were drawn.The data was collected by wintest system to evaluate the biomechanical parameters of rabbit corneal tissue. ResultsThe maximum distortion intension of rabbit cornea was (7.7432±0.6099)MPa.After three cyclic loading,the stress gradually attenuated and the stress and strain flattened as the time change with the relaxation rate 30.33％.The deformation of the specimens enhanced with time decrease with the creep rate 24.33％. ConclusionsThe biomechanical characteristics of normal rabbit cornea are revealed in this study,which offer the basis for the experimental research of rabbit model aimed at corneal disease.

Background Posterior capsular opacification (PCO)affects the pseudoaccommodation of 1CU accommodative intraocular lens (1CU AIOL).At present,few studies on the effect of PCO and Nd∶ YAG laser capsulotomy on intraocular shifting of 1CU AIOL are published.Objective The present study was to evaluate the effect of PCO and Nd∶YAG laser capsulotomy on the shifting of 1CU AIOL.Methods A respective serial caseobservational study was designed.Written informed consent was obtained from each patient prior to this study.Twentyfour eyes of 20 patients with PCO after phacoemulsification and implantation of 1CU AIOL were included in this study.Ocular examination was performed 3 months after IOL implantation,1 day before Nd:YAG laser capsulotomy and 3 months after Nd∶YAG laser capsulotomy to evaluate the distance corrected near visual acuity(DCNVA).The difference in the anterior chamber depths before and after administering 1％ pilocarpine topical eye drops was measured with the IOLMaster to determine the intraocular shifts of the IOL.The extent of IOL shifting was compared among 3 time points to assess the factors influencing IOL accommodation after 1CU AIOL implantation.Results The shifting amplitude of 1CU AIOL was(0.44±0.21)mm 3 months after implantation of 1CU AIOL,(0.27±0.11)mm 1 day before Nd ∶ YAG laser capsulotomy,and (0.34±0.10) mm 3 months after Nd ∶ YAG laser capsulotomy,showing a significant difference among them(F=7.180,P=0.001).The shifting amplitude of 1CU AIOL significantly declined 1 day before Nd∶YAG laser capsulotomy in comparison with 3 months after implantation of 1 CU AIOL(P =0.006).The shifting amplitude 3 months after Nd∶YAG laser capsulotomy increased slightly in comparison with 1 day before Nd∶YAG laser capsulotomy(P=0.059).DCNVA was(3.1±0.9)J 3 months after implantation of 1CU AIOL,(6.2±0.8) J 1 day before Nd ∶ YAG laser capsulotomy and(3.4±0.7) J 3 months after Nd ∶ YAG laser capsulotomy,with a significant difference among them (F =110.270,P =0.000).DCNVA was lower 1 day before Nd∶ YAG laser capsulotomy than 3 months after implantation of 1CU AIOL(P＜0.05).However,DCNVA was higher 3 months after Nd∶YAG laser capsulotomy than that of 1 day before Nd∶YAG laser capsulotomy (P＜0.05).There was no significant correlations between DCNVA and IOL movement 3 months after IOL implantation,1 day before Nd∶ YAG laser capsulotomy and 3 months after Nd ∶ YAG laser capsulotomy (r1 =-0.150,P1 =0.486,r2 =-0.320,P2 =0.122,r3 =-0.100,P3 =0.633).Conclusions The shifting amplitude of 1CU AIOL markedly declines due to PCO.No clinically significant influence of Nd ∶ YAG laser capsulotomy on the shifting amplitude of 1 CU AIOL is found.DCNVA can improve after Nd∶YAG laser capsulotomy.Multiple inter-related factors concerning pseudophakic accommodation may influence DCNVA.

At the beginning of the 1980s, a concept of viable but non-culturable(VBNC) was suggested. VBNC is a survival strategy adopted by microorganisms when they are exposed to environmental stress. This article try to make a summary of research of the conditions of VBNC formation, recovery of culturability and methods of VBNC cells detection. In addition, introduces the first growth factor of microorganisms-Rpf.

Objective: To investigate the chemical constituents of the flowers of Gentiana dahurica. Methods: All compounds were isolated and purified by silica gel, Sephadex LH-20, ODS, and MCI column chromatography. Their structures were determined by physicochemical properties and spectral data. Results: Twenty compounds were isolated from the flowers of G. dahurica. Among them, eight triterpenoids were identified as roburic acid (1), 3β-acetoxy-28-hydroxy-12-ene-oleanane (2), 28-hydroxy-α-amyrin (3), 28-hydroxy-β-amyrin (4), α-amyrin (5), β-amyrin (6), ursolic acid (7), and oleanolic acid (8). Twelve flavonoids were identified as 1-hydroxy-3,7,8-trimethoxyxanthone (9), kaempferol (10), naringenin (11), apigenin (12), luteolin (13), (2S)-naringenin-7-O-β-D- glucopyranoside (14), apigenin-7-O-β-D-glucopyranoside (15), luteolin-7-O-β-D-glucopyranoside (16), isoorientin (17), isovitexin (18), saponarin (19), and lutonarin (20). Conclusion: Seventeen compounds 1-11, 13-16, 19, and 20 are found from flowers of G. dahurica for the first time; Compounds 2-7, 9-11, 13-16, 19, and 20 are isolated from this species for the first time.

Objective:To investigate the relationship between T/N take-up value and the clinic feature of breast cancer, and explore the predictive ability of T/N take-up value for breast cancer prognosis.Methods:Breast cancer patients treated at Zhongshan Hospital, Fudan University and undergoing BSGI examination before surgery from Jan 2014 to Nov 2018 were collected. The relationship between T/N take-up value and the clinical features of breast cancer and DFS was analyzed.Results:A total of 419 breast cancer patients were collected. The median follow-up time after operation was 34 months. At the end of the follow-up, 43 cases relapsed and 376 cases did not. The average T/N values of CC position and MLO position before surgery were 2.74 and 2.58. The T/N value of CC is related to tumor stage (χ 2=22.077, P<0.05), lymph node metastasis ( Z=2.138, P<0.05) and degree of invasion ( Z=3.371, P<0.05). The T/N value of MLO is related to tumor stage (χ 2=23.091, P<0.05), lymph node metastasis ( Z=2.531, P<0.05) and degree of invasion ( Z=2.99, P<0.05). The best cut-off values of CC position and MLO position calculated by ROC curve are 2.59 and 2.97. Univariate analysis showed that tumor staging ( HR: 2.039, 95% CI: 1.404-2.962, P=0.001), T/N value at CC position ( HR: 4.349, 95% CI: 2.141-8.883, P<0.001) and T/N value at MLO position ( HR: 2.767, 95% CI: 1.520-5.039, P<0.001) is an independent risk factor for the prognosis of breast cancer. Multivariate COX regression analysis showed that tumor stage ( HR: 1.959, 95% CI: 1.302-2.946, P=0.001) and T/N value at MLO position ( HR: 3.498, 95% CI: 1.531-7.992, P=0.003) are independent risk factor of breast cancer prognosis. Conclusion:The BSGI T/N value has a certain correlation with breast cancer DFS.

Na+/H+ antiporter plays an important role in mechanisms of the plant salt tolerance, it extrudes Na+ from cell energized by the proton gradient generated by the plasm membrane H(+)-ATPase and/or compartmentalizes Na+ in vacuole energized by the proton gradient generated by the vacuolar membrane H(+)-ATPase and H(+)-PPiase. This review mainly discusses the latest progress in the study of Na+/H+ antiporter in plant and yeast at molecular level.
Phylogeny ; Plants ; metabolism ; Salts ; metabolism ; Sodium ; metabolism ; Sodium-Hydrogen Exchangers ; classification ; metabolism ; Vacuoles ; physiology

ObjectiveTo discuss the relationship between the cross-reactivity of antibody against the hypervariable region 1 ( HVR1 ) of hepatitis C virus and early viral response ( EVR ) in patients undergoing antiviral therapy.MethodsBy ELISA and HCV HVR1 antibody cross-reactivity matrix reagent,the differences of anti-HCV hypervariable region antibody were tested in baseline serum from 46 patients with chronic hepatitis C before antiviral therapy.HCV genotyping and HCV RNA were analyzed by RT-PCR method.The HCV RNA assay was done at three time points:before treatment,pegylated interferon in combination with ribavirin therapy for 12 and 48 weeks.ResultsIn 46 cases of chronic hepatitis C patients,there were 16 cases with HCV 2a type,30 cases with l b and 33 patients obtained EVR.The EVR incidence of type 2a[ 93.8％ ( 15/16 ) ] was higher than that of type 1 b[ 60.0％ (18/30),x2 =4.316,P ＜ 0.05 ].In the EVR group of type 1b chronic hepatitis C patients,the positive number of average multi-target HVR1 antigen was ( 12 ± 4),which was significantly higher than that in the Non-EVR patients [ (7 ± 5 ),t =2.797,P ＜0.01 ].Bv the Fisher exact test,it was showed that patients'serum response to HVR1 antigens numbered 001,003,009,013,016 were higher in EVR group than those in non-EVR group,with statistically significant (P ＜ 0.05 ).ConclusionThe cross-reactivity of HVR1 antibody may play an important role in predicting the effectiveness of antiviral therapy.

NDV strain ZJ1 strain , a highly virulent NDV strain, has been prevalent among the waterfowls in China mainland in the past years. Multi-basic amino acid sequence distribute in the protease cleavage site of F protein of this strain. Recombinant expressing plasmid pCI-FT, was generated by converting multi-basic amino acid sequence of 112, 115, 117 of the protease cleavage site of F_ 0 protein, to the non-basic amino acid sequence characteristic of avirulent NDV strain. The result from co-expression of mutant or parental F protein with homologous HN protein in COS-1 cells revealed that both mutant and parental F protein had fusion activity. The result from co-expression of mutant or parental F protein with homologous HN protein in CEF cells showed that the cleavage activity of mutant F protein was significantly reduced. The study built a foundation for mutagenesis of amino acid sequence of the protease cleavage site of F_ 0 protein at the full-length cDNA clone level, study on factors contributing to virulence and construction of candidate vaccine strain, and so on.