1.Construction and Identification of Recombinant Plasmid of p3XFLAG-CMV7-NICD1
Journal of China Medical University 2015;(3):217-220
Objective To construct the eukaryotic expression vector of Notch1intracellular domain,p3XFLAG?CMV7?NICD1,so as to prepare for the further research and exploration of effect of Notch1 on promoting epithelial?mesenchymal transition of human lens epithelial cells. Methods The cDNA fragment was reversely transcribed by RT?PCR from total RNA extracted from the SRA01/04 cells and was encoded with the specific am?plification?targeted NICD1was obtained from the SRA01/04 cells,then the cDNA fragment was inserted into p3XFLAG?CMV7 to transcribe Esche?richia coli DH5α. And the recombinant plasmid was extracted after bacterial screening by LB plating medium and confirmed by the restriction endo?nuclease digestion and DNA sequencing. Results The target gene obtained had the same molecular size as predicted. It was indicated that recom?bined p3XFLAG?CMV7 plasmid contained correct recombinant human Notch1 sequences and p3XFLAG?CMV7?NICD1 was constructed success?fully. The western blotting showed protein NICD1 expressed in SRA01/04 cells transfected with p3XFLAG?CMV7?NICD1. Conclusion The suc?cessful construction of p3XFLAG?CMV7?NICD1 will provide a foundation for a further study studies in on the effect relationship of Notch signaling pathway and in posterior capsular opacification(PCO)after cataract extraction.
2.Resveratrol inhibits high glucose-induced hypertrophy in myocardial cell line H9C2
Xiao WEI ; Jianming LEI ; Jingwen GUO ; Hua XIAO
Basic & Clinical Medicine 2017;37(3):320-324
Objective To investigate the effect of resveratrol on high glucose-induced oxidative stress and cardiomyo-cyte hypertrophy and possible mechanism .Methods H9C2 cardiomyocytes were divided into normal glucose group , high glucose group , resveratrol group and resveratrol +DAPT group respectively .The cell surface was measured by rhodamine-labeled phalloidin .Reactive oxygen species generation was measured using DCFH-DA.The contents of SOD and MDA were evaluated by colorimetry .The mRNA and protein levels of Notch 1, Hes1, ANP and BNP were evaluated by RT-qPCR and western blot .Results High glucose exposure significantly increased cell surface area ( P<0.01) and enhanced ROS and MDA generation ( P <0.01 ) with concomitant decrease of SOD activity ( P <0.01).It also decreased the mRNA and protein expressions of Notch 1 and Hes1(P<0.05), and increased the ex-pression levels of ANP and BNP ( P<0.05 ) .Resveratrol treatment inhibited cell surface expanding ( P<0.01 ) and decreased ROS and MDA generation (P<0.01) with concomitant enhance SOD activity (P<0.01), as well as in-creased the expression level of Notch 1 and Hes1 ( P<0.05 ) , and decreased the expression of ANP and BNP ( P<0.05).Conclusions Resveratrol may inhibit high glucose induced H9C2 cardiomyocytes hypertrophy by improving oxidative stress and activating Notch 1/Hes1 signaling pathway .
3.Changes of aryl hydrocarbon receptor in cardiac hypertrophy induced by high glucose in vitro
Xuejiao TANG ; Hua XIAO ; Lei ZHANG ; Xiao WEI ; Jianming LEI ; Jingwen GUO
Chinese Journal of Pathophysiology 2016;32(10):1744-1749
AIM:To investigate the changes of aryl hydrocarbon receptor (AhR) in the process of cardiomyo-cyte hypertrophy induced by high glucose , and to explore its potential mechanisms .METHODS: The rat cardiomyocytes (H9c2 cells) were divided into normal glucose group , high glucose group, DMSO group and resveratrol (an AhR antago-nist) group.The content and distribution of AhR were observed with immunofluorescence staining .The myocardial cells were stained with rhodamine-labeled phalloidin to visualize cytoskeleton , and the cell surface area were determined after im-aging by fluorescence microscopy .The generation of reactive oxygen species ( ROS) in the cardiomyocytes was measured u-sing a fluorescent probe DCFH-DA.The mRNA expression of AhR , CYP1A1, atrial natriuretic peptide ( ANP) and brain natriuretic peptide ( BNP) were evaluated by real-time quantitative PCR ( RT-qPCR).The protein levels of AhR, CYP1A1, ANP and BNP were assessed by Western blot .RESULTS:AhR was constitutively presented in the cytosol un-der normal-glucose condition and was translocated to the nuclei under high-glucose condition .High glucose induced cardiac hypertrophy , and increased ROS generation .Significant reductions in the cell size and ROS generation were observed after treated with resveratrol.The expression of AhR, CYP1A1, ANP and BNP at mRNA and protein levels in high glucose group was increased as compared with normal glucose group and resveratrol group , and the above-mentioned indexes signifi-cantly decreased in resveratrol group as compared with DMSO group .CONCLUSION: High glucose-induced cardiac hy-pertrophy increases AhR expression , which may be involved in the maintenance of glucose homeostasis in the cardiomyo-cytes.AhR translocation to the nucleus induced by high glucose results in the increases in CYP 1A1 expression and ROS generation, which may be an important mechanism of high glucose-induced cardiomyocyte hypertrophy .
4.Clinical application of lower cross-lip flap in treatment of upper lip defect
Jingjian WEI ; Jingqian XIAO ; Zhengyu LIU ; Bocheng LEI ; Xukai WANG
Chinese Journal of Medical Aesthetics and Cosmetology 2013;19(5):334-336
Objective To evaluate the clinical effect of the upper lip defect with lower cross-lip flap (Abbe-Estlander flap).Methods A total of 68 cases of upper lip defect underwent the reconstruction with cross-lip flaps.We applied Abbe flap in upper lip defect in the center,and Estlander flap in lateral upper lip defect.This method was a two-stage procedure:the first stage was to rotate flap 180 degrees into the upper lip defect and to suture with the created defect of the upper lip,with careful maintenance of blood supply from the pedicle; the second was to undertake the division of bridged pedicle and paid more attention to creation of the vermilion border.Results The flaps survived in all cases.Evaluation from 3 to 12 months after the operation showed that the shape of lips were obviously repaired with excellent aesthetic and functional results.Conclusions Abbe-Estlander flap could reconstruct anatomical features and function of the lip precisely.It seems that within certain limits (probably between one-third and one-half of total upper lip length),this flap appears to be the preferred method for upper lip reconstruction.
6.Aortic surgery through upper hemisternotomy
Lei CHEN ; Dong LI ; Wei JIANG ; Cangsong XIAO
Chinese Journal of Thoracic and Cardiovascular Surgery 2021;37(1):10-13
Objective:To summarize the feasibility and clinical experience of various types of aortic surgery through upper hemisternotomy.Methods:From June 2016 to October 2019, 63 patients underwent various types of aortic operations through upper hemisternotomy in our department. Among them, there were 51 males and 12 females with an average age of(49.7±12.7) years. All kinds of major vascular operations include: 33 cases of aortic dissection procedure; 9 cases of Bentall procedure; 7 cases of wheat procedure; 2 cases of Ross procedure; 2 cases of David procedure; 2 cases of simple ascending aorta replacement procedure; 3 cases of redo thoracic Bentall procedure; 2 cases of redo thoracic wheat operation; 1 case of redo thoracic aortic root leakage repair procedure; 2 cases of redo thoracic ascending aorta procedure, innominate artery, left common carotid artery replacement and removal of intravalvular vegetation procedure; 1 case of Bentall plus pulmonary valve and pulmonary artery replacement procedure. Among them, aortic dissection included 11 cases of ascending and total arch replacement plus descending aortic stent implantation procedure; 2 cases of AVR plus ascending and total arch replacement procedure; 5 cases of ascending and semi-arch replacement procedure; 4 cases of David plus ascending and total arch replacement plus descending aortic stent implantation procedure; 1 case of redo thoracotomy David plus ascending and total arch replacement with descending aortic stent implantation procedure; 1 case of David plus ascending and total arch replacement procedure; 6 case of AVP/AVR/Bentall plus ascending and total arch replacement and descending aortic stent implantation procedure; 2 cases of redo total arch replacement and descending aortic stent implantation procedure and aortic root vascular graft wrapping plus ascending aorta and 1 case of total arch replacement with descending aortic stent implantation procedure.Results:All 63 cases of operation were successfully completed without transforming to middle sternotomy; 1 case with acute renal failure was treated by dialysis, and then complicated with septic shock and intracranial hemorrhage, and died of ineffective rescue; 2 cases were treated by thoracotomy exploration and hemostasis; 9 cases with pericardial effusion needed pericardiocentesis; 2 cases with lower extremity muscle weakness after operation: one treated by drainage of spinal fluid and the muscle strength of the lower extremities was restored, another one complicated with infection, acute renal failure and hypoxemia was treated by anti-infection, continuous CRRT, gamma globulin infusion and rehabilitation exercise, the muscle strength of the lower extremities was eventually restored. Except for one death in hospital, all the other patients were cured and discharged.Conclusion:Well exposure of aortic root, ascending aorta and descending part of arch can be obtained through upper hemisternotomy and various types of aortic proceduer can be done with satisfactory results through this approach.
7.The Observation of Clinical Effect of Using Ambroxol in the Treatment of Acute Exacerbation of Chronic Obstructive Pulmonary Disease
Weihua HU ; Wei XIAO ; Zhenping CHEN ; Lei WANG
Journal of Kunming Medical University 2016;37(8):65-67
Objective To compare the efficacy of two doses of ambroxol in the treatment of the elderly patients with acute exacerbation of chronic obstructive pulmonary disease (COPD).Methods A hundred and ten cases of the elderly patients with an acute exacerbation of COPD were divided into low-dose group and high-dose group.Low-dose group (56 cases) were given Ambroxol 30 mg,2 times per day for 10 day and high-dose group (54 cases) were given Ambroxol 60 mg,2 times per day for 10 days.Results The effective rate in high-dose group was significantly higher than that in low-dose group.Conclusion High-dose ambroxol has significant effect in the treatment of acute exacerbation of chronic obstructive pulmonary disease.
8.Expression and significance of aryl hydrocarbon receptor in atrial tissues of patients with atrial fibrillation
Jianming LEI ; Hua XIAO ; Qiang LI ; Xiao WEI ; Jingwen GUO ; Minghan XIAO
Chinese Journal of Pathophysiology 2017;33(5):826-831
AIM:To investigate the expression of aryl hydrocarbon receptor (AhR) in atrial tissues of the patients with rheumatic heart disease (RHD), and the effects of AhR on rheumatic atrial fibrosis.METHODS:Right atrial specimens obtained from the patients with RHD requiring valve replacement surgery were divided into chronic atrial fibrillation (RHD+cAF, n=11) group and sinus rhythm (RHD+sinus rhythm, n=25) group.The patients with congenital heart disease (CHD) and sinus rhythm (CHD+sinus rhythm, n=12) who underwent heart surgery served as controls.The collagen volume fraction in the atrial specimens was examined by Masson`s trichrome staining.The protein expression and distribution of AhR, AhR nuclear translocator (ARNT) and CYP1A1 were detected by the methods of immunohistochemistry and Western blot.The mRNA expression of AhR, ARNT and CYP1A1 was detected by real-time fluorescence quantitative PCR.RESULTS:Compared with CHD+sinus rhythm group, the collagen content and the expression of AhR, ARNT and CYP1A1 were significantly increased in RHD+sinus rhythm group and RHD+cAF group.Compared with RHD+sinus rhythm group, the collagen content and the expression of AhR, ARNT and CYP1A1 were significantly increased in RHD+cAF group (P<0.05).CONCLUSION:The expression of AhR is correlated with the degree of fibrosis.The expression of AhR/ARNT/CYP1A1 is increased in atrial tissues of patients with RHD, suggesting that AhR/ARNT/CYP1A1 should be involved in atrial fibrosis of the patient with RHD.
9.HPLC enantioseparation, absolute configuration determination and anti-HIV-1 activity of (±)-F18 enantiomers.
Lei-lei ZHANG ; Hai XUE ; Li LI ; Xiao-fan LU ; Zhi-wei CHEN ; Gang LU
Acta Pharmaceutica Sinica 2015;50(6):733-737
Racemic (±)-F18 (10-chloromethyl-11-demethyl-12-oxo-calanolide A), an analog of nature product (+)-calanolide A, is a new anti-HIV-1 nonnucleoside reverse transcript inhibitor (NNRTI). A successful enantioseparation of (±)-F18 offering (R)-F18 and (S)-F18 was achieved by a chiral stationary phase prepared HPLC. Their absolute configurations were determined by measurement of their electronic circular dichroisms combined with modem quantum-chemical calculations. Further investigation revealed that (R)-F18 and (S)-F18 shared a similar anti-HIV activities, however, (R)-F18 was more potent than (S)-F18 against wild-type virus, K101E mutation and P225H mutation pseudoviruses.
Anti-HIV Agents
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chemistry
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Chromatography, High Pressure Liquid
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HIV-1
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drug effects
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Pyranocoumarins
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chemistry
10.Microscopic observation on mycorrhiza of rare herb Dysosma versipellis.
Xiao-Ming TAN ; Li-Ying YU ; Ya-Qin ZHOU ; Xiao-Lei ZHOU ; Ying WEI
China Journal of Chinese Materia Medica 2013;38(23):4044-4046
Endophytic fungi played an important role in the growth of its host plant. To investigate the mycorrhizal characteristics and the distribution of fungi in the root, an endangered wild plant-Dysosma versipellis was collected and observed by electron microscope. The results showed that the host was closely associated with endophytic fungi. The fungi were mainly distributed in the epidermis and cortex. The aseptate and septate fungi with swollen hyphae were observed in some cell of the cortex. The result provides a reference for the study of mycorrhizal structure of Dysosma genus and the interaction between the fungi and its host.
Berberidaceae
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microbiology
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ultrastructure
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Endangered Species
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Endophytes
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physiology
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ultrastructure
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Fungi
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physiology
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ultrastructure
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Microscopy, Electron
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Plant Roots
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microbiology
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ultrastructure