1.miR-335 regulate cell proliferation by targeting survivin in breast cancer cells MCF-7
Yajuan HUANG ; Feng DU ; Lingli WEI
Chongqing Medicine 2016;45(27):3753-3756
Objective To evaluate the effect of miR‐335 regulate cell proliferation by targeting survivin in breast cancer cells MCF‐7 .Methods (1)Chose breast cancer tissue and para‐carcinoma tissue ,and used RT‐PCR or Western blot to detect the expres‐sion of miR‐335 and survivin protein .(2)Chose breast cancer cells MCF‐7 ,respectively transfected miR‐335 mimic and mimic con‐trol .The expression of miR‐335 and survivin protein were detected by RT‐PCR or Western blot .(3)Chose breast cancer cells MCF‐7 ,respectively co‐transfection wild type survivin 3′‐UTR(survivin‐wt)or mutant type urvivin 3′‐UTR(survivin‐Mut)and miR‐335 mimic or negative control(NC)to breast cancer cells MCF‐7 .The cell luciferase activity were detected by dual‐luciferase report gene experiment .(4)Chose breast cancer cells MCF‐7 ,respectively transfected mimic control ,miR‐335 mimic and miR‐335 mimic+ sur‐vivin .The cell proliferation activity of each group were detected by MTT method .Results (1)Compared with para‐carcinoma tis‐sue ,the miR‐335 expression of breast cancer tissue significantly decreased(P<0 .05) ,and the survivin protein expression of breast cancer tissue significantly increased(P<0 .05) .(2)Compared with transfection mimic control ,transfection miR‐335 mimic can made the miR‐335 expression of MCF‐7 increased(P<0 .05) ,and made the survivin protein expression of MCF‐7 decreased(P<0 .05) . (3)Compared with transfection NC ,co‐transfection miR‐335 mimic and survivin‐wt can made luciferase activity of MCF‐7 signifi‐cantly decreased(P<0 .05) ,but the luciferase activity of MCF‐7 was not significantly changes after co‐transfection miR‐335 mimic and survivin‐Mut (P>0 .05) .(4)Compared with transfection mimic control ,the MCF‐7 proliferative activity significantly decreased after transfection miR‐335mimic(P<0 .05) .Compared with transfection miR‐335 mimic ,the MCF‐7 proliferative activity signifi‐cantly increased after transfection miR‐335 mimic+survivin(P<0 .05) ,but still lower than transfection mimic control(P<0 .05) . Conclusion In breast cancer tissue ,the miR‐335 show low expression ,and survivin show high expression .miR‐335 can target sur‐vivin to inhibit the proliferation activity of breast cancer cells MCF‐7 .
2. Optimization of response surface of fresh-cut processing technology in Corydalis Rhizoma producing area
Chinese Traditional and Herbal Drugs 2019;50(5):1111-1116
Objective To optimize the fresh-cutting process of Corydalis Rhizoma by response surface methodology, and obtain the best technology for fresh-cut processing of Corydalis Rhizoma. Methods The slice thickness, the hot air drying temperature and the loading amount were taken as the investigation factors. The total alkaloid content, the extract content and the drying efficiency were taken as the indicators. The weight coefficient of each index was obtained by principal component analysis method, and the comprehensive score was calculated. Single factor analysis was performed on the investigation factors to obtain the initial optimization range, and the response surface optimization method was used to optimize the final process optimization parameters. Results The best production conditions for fresh-cut processing were determined as slice thickness of 4 mm, dry load of 7 kg/m2, drying temperature of 85 ℃, total drying time of (279.0 ± 1.1) min, and comprehensive score of (0.860 6 ± 0.010 0). At this time, the total alkaloid content of Corydalis Rhizoma was (6.274 ± 0.030) mg/g, the content of the leachate was (17.86 ± 0.22)%, and the drying efficiency was (25.09 ± 0.00) g/(m2∙min). Conclusion The established drying process can better preserve the content of alkaloids and extracts in Corydalis Rhizoma, maintain high drying efficiency, ensure high quality and reduce energy consumption of enterprises, and provide a reliable theoretical basis for the production and processing of Corydalis Rhizoma pieces.
3.Progress in the study of near-infrared fluorescent probes for the detection of β-amyloid deposition in Alzheimer's disease.
Lei DU ; Hai-wei FENG ; Yu-yan LI
Acta Pharmaceutica Sinica 2015;50(5):528-534
Alzheimer's disease (AD) is the most common cause of cognitive impairment in older people. With the aging of society is more and more serious, AD caused great burden to patients and society. A β is a classical biomarker of AD, which has been widely used in clinical diagnosis of AD patients. Compared with positron emission tomography (PET) and single photon emission computed tomography (SPECT), near infrared fluorescence imaging has many advantages including highly sensitive, non-invasive, safety and inexpensive. Therefore, many research groups have focused on developing the molecular probes of near-infrared fluorescence (NIRF) imaging. In this article, we will review the progress of the probes of NIRF.
Alzheimer Disease
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diagnosis
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Amyloid beta-Peptides
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analysis
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Fluorescence
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Fluorescent Dyes
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Humans
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Molecular Probes
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Positron-Emission Tomography
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Spectroscopy, Near-Infrared
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Tomography, Emission-Computed, Single-Photon
4.Comparative Study on Early and Long-term Outcomes Between On-pump and Off-pump Coronary Artery Bypass Grafting in Elder Female Patients
Jiqiang ZHANG ; Heng ZHANG ; Junzhe DU ; Wei FENG
Chinese Circulation Journal 2015;(12):1143-1146
Objective: To compare the early and long-term outcomes between on-pump and off-pump coronary artery bypass grafting (CABG) in elder female patients.
Methods: A total of 763 female patients elder than 65 years of age received isolated CABG in our hospital from 1999-01 to 2008-12 were retrospectively studied. The patients were divided into 2 groups according to operational method: On-pump group,n=331 and Off-pump group,n=432. The mortality at 30 days post-operation, in-hospital clinical indexes and long term mortality with MACCE as all cause death, myocardial infarction (MI), stroke and repeated revascularization were compared between 2 groups.
Results: Compared with On-pump group, the patients in Off-pump group had the elder age (P<0.01), with the higher rate of cerebral vascular accident (P=0.023); less family history of coronary artery disease (P=0.012), angina (P<0.001) and emergent operation (P=0.015), less venous distal anastomoses (P<0.001) ; while higher incomplete revascularization (P<0.001). Logistic regression model comparison indicated that Off-pump group had the lower mortality at 30 days post-operation (P=0.038), less application of blood products (P<0.001), lower rate of >24h mechanical ventilation (P<0.001), less renal failure (P=0.022), pulmonary complications (P<0.001) and re-exploration for bleeding (P=0.021). The morbidity of all post-operative complications were similar between 2 groups (P=0.110). Cox regression model comparison showed that long-term all cause mortality (P=0.477) and occurrence of MACCE (P=0.265) were similar between the two groups.
Conclusion: Off-pump CABG would reduce the mortality at 30 days post-operation, have less application of blood products, shorter post-operative mechanical ventilation, less early post-operative renal failure, pulmonary complications and re-exploration for bleeding. While it could not reduce the long-term mortality and MACCE occurrence.
5.The phylogenetic analysis of hepatitis C virus genotype 1b mutants in China
Rui ZHANG ; Guohua QIU ; Feng LIU ; Shaocai DU ; Lai WEI
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objectives In our studies before, we reported mutation C-T at -222 of hepatitis C virus type 1b genotyped on 5′ noncoding region (5′NCR). There was no this mutation of type 1b recorded in GenBank. Since genotypiog based on 5′NCR cannot differentiate subtypes, it is necessary to investigate whether the hepatitis C virus genotype 1b strains with mutation C-T belongs to other subtypes of genotype 1. Methods 64 HCV genotype 1b samples were amplified from 5′NCR by RT-PCR.Then the products were digested by use of BamHⅠ restriction enzymes. We randomly chose 6 samples with BamHⅠ restriction site and subjected them to amplification of 5′NCR and NS5B. The sequences of 5′NCR were analyzed. Sequences of NS5B fragments from the 6 samples were respectively subjected to phylogenetic analysis with subtypes of genotype 1 and 38 complete genomes of genotype 1b from GenBank.Results The phylogenetic analysis of 6 samples and subtypes of genotype 1 indicated that the genotypes of 6 strains with BamHⅠ restriction site in 5′NCR were type 1b, instead of subtype of type 1. Tree construction with 38 complete genomes of genotype 1b showed that the 6 mutants of genotype 1b did not belong to the same branch of the tree and there were no genetic differences between the mutants and the other strains of genotype 1b.Conclusions Our research indicated that the hepatitis C virus stains of C-T in 5′NCR were mutants of type 1b. Since the 5′NCR is a highly conserved region of HCV, the mutation might have some relationship to the long-time IFN therapy.
6.The genetic heterogeneity of 5’-NCR of hepatitis C virus genotype 1b
Shaocai DU ; Guohua QIU ; Feng LIU ; Rui ZHANG ; Lai WEI
Chinese Journal of Laboratory Medicine 2001;0(02):-
Objective To investigate the genetic heterogeneity of 5’-NCR of HCV genotype 1b.Methods We selected 147 HBV genotype 1b serum samples. HCV 5’-NCR fragments were amplified from these samples by use of RT-PCR assay and sequenced after using restriction endonuclease Mbo Ⅰ and BamH Ⅰ. We analyzed the phylogenetic trees of the samples and compared them with 40 isolates of HCV genotype 1b.Results The sequencing reports indicated the isolates of HCV genotype 1b recognized by BamHⅠat position 117 by a substitution (C-T) at position 120 in HCV genotype 1b viruses in China. Of 147 samples, 17 (11.56%) samples for one BamHⅠrecognization position, 26 (17.69%) for neither BamH Ⅰ recognization position nor Mbo Ⅰ,6 samples (4.08%) for two Mbo I recognization position, 18(12.24%)for one MboⅠrecognization position.Conclusions Of 147 HBV genotype 1b serum samples, 54.42% for one Mbo I recognization position, whether this have relationship with the response to IFN therapy provide the framwork for future detailed investigation of HCV antviral therapy. There is specific BamH I recognization position in isolates of HCV genotype 1b in China as compared to the other 40 HCV 1b isolates. The role of this specific mutation needs to be further researched.
7.Study on the expression,purification and bioactivity of recombinant T?16
Xiuping FENG ; Wei MA ; Bairong DU ; Dongmei YAN ; Xun ZHU
Chinese Journal of Immunology 1986;0(04):-
Objective:To study the bioactivity of thymosin ?16 (T?16) in vitro and in vivo.Methods:Recombinant His-SUMO-T?16 was constructed and transformed into E.coli BL21(DE3) for induced expression.The product was treated by ultrasonication,ion-exchange chromatography and metal chelation chromatography respectively for purification.The fusion protein was cut by His-SUMO protease and then further purified by metal chelation chromatography and Superdex 30 gel chromatography.Results:Recombinant fusion protein His-SUMO-T?16 was soluble,whose specific activity was 5.3?105 U/mg.It could promote the proliferation of BALB/c 3T3 cells,rabbit corneal cells,and chicken embryo chorion vessels in vitro,and both the proliferation and migration of vascular endothelial cells in vitro were enhanced,and rabbit skin healing of alkali burns in vivo was accelerated.Conclusion:E.coli expressing vector of recombinant His-SUMO-T?16 fusion protein is constructed successfully,and recombinant protein T?16 has significant repairing effects.The study established a good foundation for further industrialization of T?16.
8.Plasma soluble triggering receptor expressed on myeloid cells-1 as a marker of sepsis: a meta-analysis
Du CHEN ; Shiqi LU ; Feng XU ; Wei LI
Chinese Journal of Emergency Medicine 2013;22(9):1025-1029
Objective To investigate the value of plasma soluble triggering receptor expressed on myeloid cells-1 as a diagnosis marker of sepsis.Methods Articles on plasma soluble triggering receptor expressed on myeloid cells-1 as a marker of sepsis which were public published in the PubMed,Ovid,Springer,Wanfang database from 1991-2012 were searched and conducted a meta-analysis by MetaDiSc and Stata.Results Seven articles were selected to the meta-analysis according to the inclusion criteria,of which cut-off values varied signicantly from studies.Due to the data heterogeneity (I2 > 50%,P <0.05),random model was used to pool the effect sizes.The overall combined effect sizes:sensitivity =81% (95%CI:0.76-0.86); specificity =81% (95% CI:0.76-0.86); DOR =30.03 (95% CI:7.89-114.37) ; AUC of SROC =0.905 9; Q*-0.837 6.Deek' s funnel plot showed little publication bias.Conclusions Plasma soluble triggering receptor expressed on myeloid cells-1 may be a useful adjunctive tool for the diagnosis of sepsis.However,further studies are needed in order to identify the best cut-off value in the diagnosis of sepsis.
9.The relationship between extravascular lung water and the prognosis of sepsis acute lung injury/acute respiratory distress syndrome: a Meta analysis
Du CHEN ; Shiqi LU ; Feng XU ; Wei LI
Chinese Journal of Emergency Medicine 2012;21(4):375-378
Objective To investigate the value of extravascular lung water (EVLW) in predicting the prognosis of sepsis acute lung injury (ALI) / acute respiratory distress syndrome (ARDS).Methods Relevant published articles concerned with EVLW both in Chinese and in English from 1991-2011 were searched,and Meta-analysis was carried out with Stata software. Results Seven articles were selected for Meta-analysis according to the inclusion and exclusion criteria.The results indicated EVLW of dead patients group kept in a high level,while EVLW showed a down-trend in the survival group.The difference in level of EVLW between the two groups became more obvious with the day ( SMDd1< SMDd2 < SMDd3 ) ( the 1st day:SMDd1=0.29,95%CI:0.047-0.532; the2nd day:SMDd2=1.64,95%CI:0.14-3.13; the 3rd day:SMDd3 =1.83,95% CI:0.56-3.10).Conclusions EVLW and its dynamic change in the early stage of sepsis ALI/ARDS could predict the prognosis.This suggested the sustained high level would lead to high mortality.
10.Doppler ultrasound screening for deep vein thrombosis in patients with lower limb fracture
Youqing YAN ; Hua REN ; Kun FENG ; Chaohui DU ; Zhiyu WEI
Chinese Journal of General Practitioners 2011;10(9):673-674
One hundred and sixteen patients with lower limb fracture were screened by Doppler ultrasound for deep vein thrombosis(DVT) of bilateral lower extremities within the first 72 h, d7 and d21 after fracture. Results showed that DVT was detected in 31 (26. 7% ) out of 116 cases within 72 h; at d7 and d21 DVT was detected in 3 and 1 patient respectively with a cumulated DVT rate of 30. 2% in 3 weeks.Serial Doppler ultrasonography is of value in screening for DVT of the lower extremities in patients with lower limb fracture at early stage.