2.Effects of curcumin on apoptosis and caspase-3 expression in cortex tissue in rats with hypoxic ischemic brain damage.
Guang-Mei YIN ; Lin-Sheng YU ; Shu-Zhen WU ; Guang-Hua YE
Chinese Journal of Applied Physiology 2010;26(4):504-506
Animals
;
Apoptosis
;
drug effects
;
Brain
;
drug effects
;
Caspase 3
;
metabolism
;
Curcumin
;
pharmacology
;
Hypoxia-Ischemia, Brain
;
metabolism
;
pathology
;
Male
;
Rats
;
Rats, Sprague-Dawley
3.Construction of nuclear factor of activated T-cells 5 mRNA 3'-untranslated region reporter vector and targeting verification between NFAT5 and miR-155
Bin SHU ; Wenting LI ; Zhen LIU ; Yajie ZHANG ; Bin YIN ; Pan ZHAO ; Tongwei ZHANG ; Chiyu JIA
Chongqing Medicine 2017;46(8):1009-1011,1014
Objective To construct a Luciferace reporter vector containing the 3'untranslated region (3'UTR) of NFAT5 and measure the correlation between NFAT5 and miR-155.Methods The miR-155 targeting NFAT5 3'UTR was predicted by Target Scan,Mir Base and Pic Tar.NFAT5 and mutant NFAT5 sequence(NFAT5-mu) were then designed and synthesized,and they were cloned into pMIR-REPORTTM Luciferace reporter vector.Human embryonic kidney-293AD (HEK-293AD) cells of the 4th passage were divided into 4 groups according to the random number table.cells in plasimd +miR-155 mimics groups were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 mimics;cells in plasimd + miR-155 mutated groups were transfected with pMIR-NFAT5-mu recombinant plasimid,pRL-Tk plasmid and miR-155 mimics;cells in plasimd + miR-155 control groups were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 Negative control;cells in plasimd +miR-155 inhibitor were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 inhibitor;and were respectively transfected into together by liposome.After culture for 24 h,the luciferase activity was detected by dual luciferase reporter assay system.Results TargetScan,Miranda and PicTar shared the results that NFAT5 has the complementary binding sites with 3'UTR of miR-155.And luciferase reporter vectorwas constructed.Therefore the result of sequencing and double digesting of recombined plasmid were completely correct.Dual-luciferase reporter assay showed that miR-155 possesses a target effect on 3'UTR of NFAT5.Compared to the pMIR-NFAT5 + miR-control group,the luciferase activity of the pMIR-NFAT5 + miR-1 5 5 mimics group was decreased,with statistically significant difference(P<0.01),while there was no significant difference at other time points(P>0.05).Conclusion The pMIR-NFAT5 recombinant plasmid and pMIR-NFAT5 recombinant mutated plasmid were confirmed with successful construction.and it was found that miR-155 can target NFAT5 mRNA 3'-UTR.The results provide the experiment data for further disclosing the mechanism of inhalation injury on the level of gene expression.
4.Application of peak expiratory flow rate in asthmatic children
yan-ling, YUE ; xin, WU ; zhi-gang, ZHANG ; shu-zhen, YIN
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To study the clinical application of the peak expiratory flow rate( PEFR) in children with asthma.Methods The PEFRs of 43 cases of asthma at the acute stage were measured to guide the clinical grading and therapy by the optimum individual PEFR of the patients at the remission stage, and the descending rate and warning value of individual PEFR were determined to investigate the relationship between the individual PEFR descending rate and the asthma attack conditions in the standard and nonstandard monitoring groups.Results At the asthma attack stage, the clinical symptoms became severe with PEFR declining; at the remission stage, the preventive application of drugs was based on the changes of the individual PEEF descending rate . The case number and frequency of asthma attack and the cases subjected to moderate or severe attacks in the standard group were 15 %,23.5 % and 25 % respectively; while those in the nonstandard group were 43.5 %, 75.5 % and 76.9 % respectively, which showed a significant difference( P
5.Pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation on recurrent pterygium
Ting, LI ; Shu-Xian, CHEN ; Xu-Guang, XIA ; Zheng, YIN ; Mu-Zhen, HUANG ; Ping-Ying, GUO
International Eye Science 2014;(9):1715-1716
To discuss the effective method of decreasing the postoperative recurrence rate of recurrent pterygium.
●METHODS:Totally 126 cases (126 eyes) with recurrent pterygium were randomly divided into A group (56 cases) and B group ( 70 cases ). Group A was treated by pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation, group B was treated by amniotic membrane transplantation. The followed-up time after surgery was 6-24mo.
●RESULTS:ln group A, postoperative 5-7d (average 5. 62± 1. 38d), cornea epithelium was repaired. ln group B, postoperative 7- 10d ( average 7. 38 ± 1. 12d), the corneal wound was healed. There was statistical significant difference between two groups (t = 4. 307,P<0. 05). Three cases recurrence were noted in A therapeutic group (56 cases), the recurrent rate was 5. 4%; Twelve cases recurrence were noted in B compared group (70 cases), the recurrent rate was 17. 1%. There was statistical significant difference between two groups(P<0. 05).
●CONCLUSlON: lt is suggested that pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation is effective in the treatment of recurrent pterygium.
6.Control study of early and late post-operative functional exercise in Arnold-Chiari malformation patients
Chinese Journal of Modern Nursing 2009;15(13):1222-1224
Objective To study influence of early and late functional exercise on post-operative symptomatic relief and complication in Arnold-Chiari malformation patients. Methods A total of 74 Arnold-Chiari malformation patients were divided into two groups: early functional exercise(n=41) and late functional exercise(n=33). The result of the two groups were compared. Results Age and sex of the two groups had no significant difference. Pain relief, muscle force improvement, thermesthesia improvement, numbness relief and decubitus in the two groups had significantly different. Symptoms of patients who did early functional exercise relieved better. Conclusions Early post-operative functional exercise had beneficial effect on symptom relief and complication prevention in Arnold-Chiari malformation patients.
7.Coupled exofixator for comminuted fracture of humeral shaft.
Hong-ru MA ; Shu-lin MA ; Tong-zhen YIN
Chinese Journal of Traumatology 2005;8(1):33-35
OBJECTIVETo report and assess the efficacy of a coupled exofixator in the treatment of comminuted fracture of the humeral shaft.
METHODSFrom June 1999 to September 2003, 24 patients with comminuted fracture of the humeral shaft were treated in our department, in whom 11 were involved in left humerus fractures and 13 in right humerus fractures. Closed reduction or open reduction with a small incision as well as a coupled exofixator was used to treat these patients.
RESULTSAll cases got anatomical reduction after 6-12 months follow-up. The time for fracture union averaged 5.8 months with a good functional recovery of the shoulder-elbow joints.
CONCLUSIONSThe coupled exofixator is favorable to the treatment of comminuted humeral shaft fractures. It can shorten union time and avoid nonunion occurrence.
Adolescent ; Adult ; External Fixators ; Female ; Fracture Fixation ; methods ; Fractures, Comminuted ; surgery ; Humans ; Humeral Fractures ; surgery ; Male ; Middle Aged ; Treatment Outcome
8.Effects of microRNA-146a on Fas-associated factor 2 and inflammatory factors in human lung adenocarcinoma A549 cells under the stimulation of cigarette smoke extract.
Wenting LI ; Zhen LIU ; Chiyu JIA ; Bin YIN ; Bin SHU
Chinese Journal of Burns 2016;32(2):97-104
OBJECTIVEUnder the premise of smoke inhalation injury, to explore the effects of microRNA-146a on Fas-associated factor 2 (FAF-2) and inflammatory factors in human lung adenocarcinoma A549 cells under the stimulation of cigarette smoke extract (CSE).
METHODS(1) The pMIR-FAF-2 recombinant plasmid and the pMIR-FAF-2 recombinant mutated plasmid were constructed. Human embryonic kidney 293 (HEK-293) cells of the third passage were divided into 3 groups according to the random number table, with 5 wells in each group. Cells in plasmid+ microRNA control group were transfected with pMIR-FAF-2 recombinant plasmid, pRL-TK plasmid, and microRNA control; cells in plasmid+ microRNA-146a group were transfected with pMIR-FAF-2 recombinant plasmid, pRL-TK plasmid, and microRNA-146a mimics; cells in mutated plasmid+ microRNA-146a group were transfected with pMIR-FAF-2 recombinant mutated plasmid, pRL-TK plasmid, and microRNA-146a inhibitor. After culture for 24 h, the relative luciferase activity in cells was assessed by dual-luciferase reporter gene assay. (2) Human lung adenocarcinoma A549 cells of the third passage were divided into 3 groups according to the random number table, with 4 wells in each group. Cells in microRNA control group were transfected with microRNA control; cells in microRNA-146a enhancement group were transfected with microRNA-146a mimics; cells in microRNA-146a inhibition group were transfected with microRNA-146a inhibitor. After culture for 24 h, the mRNA expression levels of microRNA-146a and FAF-2 in cells were determined with real-time fluorescent quantitative reverse transcription-PCR. (3) A549 cells of the third passage were stimulated by 0.8% CSE for 24 h after being divided and treated with the same method used in experiment (2). The mRNA expression levels of FAF-2, IL-8, monocyte chemotactic protein-1 (MCP-1), and growth-regulated oncogene-α (GRO-α) in cells were determined with real-time fluorescent quantitative reverse transcription-PCR. The protein expression levels of IL-8, MCP-1, and GRO-α in A549 cell culture supernatant were determined by enzyme-linked immunosorbent assay. The protein expression level of cyclooxygenase 2 (COX-2) of cells was assessed by Western blotting. Data were processed with one-way analysis of variance and LSD test.
RESULTS(1) The pMIR-FAF-2 recombinant plasmid and pMIR-FAF-2 recombinant mutated plasmid were confirmed with successful construction. The relative luciferase activity in HEK-23 cells of plasmid+ microRNA control group was close to that of mutated plasmid+ microRNA-146a group (P>0.05). The relative luciferase activity in HEK-23 cells of plasmid+ microRNA-146a group was significantly lower than that of plasmid+ microRNA control group and mutated plasmid+ microRNA-146a group (with P values below 0.01). (2) The expression level of microRNA-146a in A549 cells of microRNA control group was close to that of microRNA-146a inhibition group (P>0.05), and they were both significantly lower than the expression level of microRNA-146a in A549 cells of microRNA-146a enhancement group (with P values below 0.01). The mRNA expression level of FAF-2 in A549 cells of microRNA control group was close to that of microRNA-146a inhibition group (P>0.05), and they were both significantly higher than the mRNA expression level of FAF-2 in A549 cells of microRNA-146a enhancement group (with P values below 0.05). (3) After stimulation of CSE, the mRNA expression level of FAF-2 in A549 cells of microRNA control group (1.46±0.21) was close to that of microRNA-146a inhibition group (1.43±0.34, P>0.05), which were both significantly higher than the mRNA expression level of FAF-2 in A549 cells of microRNA-146a enhancement group (0.57±0.11, with P values below 0.05). The mRNA expression levels of IL-8, MCP-1, and GRO-α in A549 cells of microRNA-146a enhancement group were significantly lower than those of microRNA control group and microRNA-146a inhibition group (with P values below 0.01). The mRNA expression levels of IL-8, MCP-1, and GRO-α in A549 cells of microRNA-146a inhibition group were significantly higher than those of microRNA control group (with P values below 0.05). The protein expression levels of IL-8, MCP-1, and GRO-α in A549 cell culture supernatant of microRNA-146a enhancement group were significantly lower than those of microRNA control group and microRNA-146a inhibition group (with P values below 0.05). The protein expression level of IL-8 in A549 cell culture supernatant of microRNA-146a inhibition group was close to that of microRNA control group (P>0.05), while the protein expression levels of MCP-1 and GRO-α in A549 cell culture supernatant of microRNA-146a inhibition group were significantly lower than those of microRNA control group (with P values below 0.05). The protein expression level of COX-2 in A549 cells of microRNA-146a enhancement group was significantly lower than the levels of microRNA control group and microRNA-146a inhibition group (with P values below 0.05). The protein expression level of COX-2 in A549 cells of microRNA control group was close to that of microRNA-146a inhibition group (P>0.05).
CONCLUSIONSIn A549 cells, after being transfected with microRNA-146a and stimulated by CSE, microRNA-146a can decrease the expression of FAF-2 through integrating with the 3'-untranslated region of target gene FAF-2, thereby decrease the expression of inflammatory factors.
Adenocarcinoma ; chemically induced ; Blotting, Western ; Chemokine CCL2 ; metabolism ; Enzyme-Linked Immunosorbent Assay ; HEK293 Cells ; Humans ; Interleukin-8 ; Lung ; drug effects ; metabolism ; Lung Neoplasms ; chemically induced ; MicroRNAs ; analysis ; Plasmids ; RNA, Messenger ; Smoke ; adverse effects ; Smoking ; Transfection
9.Effect of substrate of edible mushroom on continuously cropping obstacle of Rehmannia glutinosa.
Rui-Hong RU ; Xuan-Zhen LI ; Xiao-Shu HUNAG ; Feng GAO ; Jian-Ming WANG ; Ben-Yin LI ; Zhong-Yi ZHANG
China Journal of Chinese Materia Medica 2014;39(16):3036-3041
The continuous cultivation of Rehmannia glutinosa causes the accumulation of phenolic acids in soil. It is supposed to be the reason of the so called "continuously cropping obstacle". In this study, phenolic acids (hydroxybenzoic acid, vanillic acid, eugenol, vanillin and ferulic acid) were degraded by the extracta of all the tested spent mushroom substrate (SMS) and the maximal degradation rate was 75.3%, contributed by extraction of SMS of Pleurotus eryngii. Pot experiment indicated that hydroxybenzoic acid and vanillin in soil were also degraded effectively by SMS of P. eryngii. The employment of SMS enhanced ecophysiology index to near the normal levels, such as crown width, leaves number, leaf length, leaf width and height. At the same time, the fresh and dry weight and total catalpol concentration of tuberous root weight of R. glutinosa was increased to 2.70, 3.66, 2.25 times by employment of SMS, respectively. The increase of bacteria, fungi and actinomycetes numbers in rhizosphere soil were observed after the employment of SMS by microbial counts. The employment of SMS also enhanced the enzyme activity in soils, such as sucrase, cellulase, phosphalase, urease and catelase. These results indicated that the employment of SMS alleviated the continuously cropping obstacle of R. glutinosa in some extent.
Agaricales
;
chemistry
;
metabolism
;
Agriculture
;
methods
;
Biodegradation, Environmental
;
Hydroxybenzoates
;
analysis
;
metabolism
;
Rehmannia
;
growth & development
;
metabolism
;
Soil
;
chemistry
;
Soil Microbiology
10.Cloning and tissue expression of 4-coumarate coenzyme A ligase gene in Angelica sinensis.
Sui-chao WEN ; Yin-quan WANG ; Jun LUO ; Qi XIA ; Qin FAN ; Shu-nan LI ; Zhen-heng WANG
China Journal of Chinese Materia Medica 2015;40(24):4824-4829
4-coumarate coenzyme A ligase is a key enzyme of phenylpropanoid metabolic pathway in higher plant and may regulate the biosynthesis of ferulic acid in Angelica sinensis. In this study, the homology-based cloning and rapid amplification of cDNA ends (RACE) technique were used to clone a full length cDNA encoding 4-coumarate coenzyme A ligase gene (4CL), and then qRT-PCR was taken for analyzing 4CL gene expression levels in the root, stem and root tissue at different growth stages of seedlings of A. sinensis. The results showed that a full-length 4CL cDNA (1,815 bp) was obtained (GenBank accession number: KT880508) which shares an open reading frame (ORF) of 1 632 bp, encodes 544 amino acid polypeptides. We found 4CL gene was expressed in all tissues including leaf, stem and root of seedlings of A. sinensis. The expressions in the leave and stem were increased significantly with the growth of seedlings of A. sinensis (P < 0.05), while it in the root showed little change. It indicates a time-space pattern of 4CL gene expression in seedlings of A. sinensis. These findings will be useful for establishing an experiment basis for studying the structure and function of 4CL gene and elucidating mechanism of ferulic acid biosynthesis and space-time regulation in A. sinensis.
Amino Acid Sequence
;
Angelica sinensis
;
genetics
;
Base Sequence
;
Cloning, Molecular
;
Coenzyme A Ligases
;
genetics
;
DNA, Complementary
;
chemistry
;
Molecular Sequence Data