1.Mechanism of DJ -1 against oxidative stress and its roles in the pathogenesis of age -related macular degeneration and Fuchs endothelial corneal dystrophy
Ya-Lan, ZHOU ; Jun, ZENG ; Bai-Hua, CHEN
International Eye Science 2016;16(8):1465-1468
?DJ-1 has been reported to act as aredox-activated chaperone and sensor of oxidative stress participated in a variety of activities in cellular, playing an important role in resisting oxidative stress, regulating signaling pathways and gene transcription, and maintaining mitochondria dynamic balance. DJ -1 is closely related to the occurrence and development of various diseases. Recently, the effect of DJ-1 in eye diseases has drawn more attention, and researchers have found its significant role of resistance to oxidative stress in the pathogenesis of Fuchs endothelial corneal dystrophy ( FECD) and age-related macular degeneration ( AMD ) .This review will state the mechanism of DJ-1 against oxidative stress and its role in the development of eye diseases.
2.Protective effect of epimedium flavonoids injection on experimental myocardial infarction rats.
Xiu-Lan HUANG ; Wei WANG ; Ya-Wei ZHOU
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(1):68-71
OBJECTIVETo investigate the protective effect of epimedium flavonoids Injection (EFI) on experimental acute myocardial infarction (AMI) rats.
METHODSRats were randomly divided into 6 groups, the acute myocardial infarction model was established by ligating left anterior descending branch of coronary artery (LAD). After operation, the rats in the sham-operation and model group were intravenous injected with 5% glucose injection, those in the positive medicine group were intravenous injected with nitroglycerin 0.3mg/kg, while rats in the low-, middle- and large-dose EFI group were intravenous injected with TFE in a dose of 10, 20, 40 mg/kg respectively. ECG was monitored before and after coronary artery ligation, and after treatment at different time points. At the same time, the millivolt of ST and ST-T segment were measured. The changes of serum creatine phosphokinase (CPK), lactate dehydrogenase (LDH), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were determined, and the myocardial infarcted area was detected by MTT respectively 3 h after LAD. Results After intravenous injection of EFI in a dose of 10, 20, 40 mg/kg, the myocardial infarcted area of AMI rats could be decreased in different degree, the activity of serum CPK, LDH and the content of MDA decreased (P < 0.05 or P < 0.01), while the activity of serum SOD increased significantly (P < 0.05 or P < 0.01). It could began to lower the elevated ST-T segment 5 min after medication and the action could last for 3 h (P < 0.05 or P < 0.01).
CONCLUSIONEFI has a protective effect against acute myocardial ischemia caused by LAD, and the effect is quickly initiated.
Animals ; Creatine Kinase ; blood ; Epimedium ; chemistry ; Female ; Flavonoids ; isolation & purification ; therapeutic use ; L-Lactate Dehydrogenase ; blood ; Male ; Myocardial Infarction ; drug therapy ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; blood
3.In vitro establishment of a three-dimensional hepatocyte culture system using collagen hydrogel as scaffolds
Min WANG ; Ya LAN ; Hao HU ; Yongquan SHI ; Ying HAN ; Xinmin ZHOU
Chinese Journal of Tissue Engineering Research 2013;(29):5323-5330
BACKGROUND:Rapid loss of liver-specific functions of the cultured hepatocytes limits the development of hepatocyte-based therapies. OBJECTIVE:To establish a three-dimensional culture system based on col agen hydrogel that enables to enhance liver-specific functions for a long period during culture of hepatocytes. METHODS:Hepatocytes were isolated from Sprague-Dawley rats and then encapsulated into liquid type Ⅰcol agen solution that was premixed with hepatocyte growth factor and Dulbecco's modified Eagle’s medium to create hepatocyte/col agen hydrogel compounds. The compounds were inoculated into a 96-wel plate. After gelation, culture medium was added. Light microscope, hematoxylin-eosin staining and transmission electron microscopy were used to examine the morphological characteristics and ultrastructure of the cultured hepatocytes. The cellsupernatant was col ected and tested for albumin secretion and urea synthesis. Periodic acid-schiff staining, immunofluorescence staining and quantitative real-time PCR were also used to further clarify liver-specific phenotype or function of the hepatocytes.RESULTS AND CONCLUSION:(1) Light microscope revealed that hepatocytes were round shape and distributed uniformly in col agen hydrogel. The three-dimensional hepatocyte culture system exhibited similarities to liver-like structure and tight junction were formed between hepatocytes after 14 days of culture. (2) Within the three-dimensional culture system, hepatocytes remained positive for periodic acid-schiff staining, albumin and hepatocyte nuclear factor-4αpositive after 14-day culture, which provided the convincing evidence of highly differentiated primary hepatocytes with functions of glycogen and albumin synthesis. (3) The albumin and urea productions in the three-dimensional culture system had a significantly higher level than in the two-dimensional culture, and could remain at a high level at least for 15 days. (4) The expression levels of hepatocyte-specific genes including Albumin, HNF-4α, Claudin-3, CYP1A1, CYP3A1 and G6P were significantly improved in the three-dimensional culture as compared with the two-dimensional culture. The col agen hydrogel based three-dimensional culture system provides a valuable model to enhance the hepatocyte functional maintenance and lay the foundation for the development of hepatocyte-based therapy for liver disease.
4.Effect of magnitude and duration on the performance of Cumulative Sum
Ding-Lun ZHOU ; Wei-Zhong YANG ; Ya-Jia LAN ; Zhong-Jie LI
Chinese Journal of Epidemiology 2012;33(6):617-621
To explore the effect of magnitude and duration on the performance of Cumulative Sum (CUSUM),with simulation method used on the subject after the insertion of 11 outbreak events into baseline data with Poisson distribution.Sensitivity fluctuated from 9.1% to 100.0% with specificities higher than 98.6%.Sensitivity was significantly correlated with magnitude,and increased along with the increase of magnitude.However,no significant correlation was observed between sensitivity and duration.A magnitude which was at least 2.6 times higher than that of the mean daily baseline could result in the sensitivity of 100.0%.Time-lag would be improved along with the increase of magnitude.Time between onset and detection of an outbreak was no longer than one day when magnitude was more than 1.8 of the mean daily baseline.In summary,the performance of CUSUM was influenced by magnitude,but not by duration.CUSUM had the advantage of good time-lag and high sensitivity when the outbreak magnitude was more than 2.4 time over the baseline data.
5.Immunoreactivity Studies of Various HEV Antigen with Anti-HEV IgM of the Sear from Patients Infected with Hepatitis E Virus
Guang-ya, RONG ; Ji-wen, ZHOU ; Li, LEI ; Gui-lan, ZHAO ; Jie, SUN
Virologica Sinica 2001;16(2):128-130
Objective To compare the Immunoreactivity of various HEV Antigen with Anti-HEV IgM. Methods Solid-phase enzyme immunoassay( EIA ) was developed for detecting anti-HEV IgM by using synthetic peptides E30, E42, E33, and recombinant antigen from HEV ORF-2. Results Of 60 anti-HEV positive sera by using E30, E42, E33 and recombinant antigen as coating antigen, Anti-HEV IgM positive rates were 76.7%, 26.6%, 18.3% and 66.7% respectively. In Acute-phase and convalescence-phase sera of the patients with Hepatitis E, Anti-HEV IgM positive rate was 90% and 3.3% respectively. Conclusions The HEV E30-based EIA will be very useful in the early diagnosis of Hepatitis E.
6.Clinical Aspects and Treatment of Enuresis Companied with Spina Bifida Occulta in Children
ya-lan, LIU ; fei-qiu, WEN ; ke-ying, ZHOU ; xiao-yuan, ZHANG
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To investigate the clinical states of enuresis children companied with spina bifida occulta(SBO)and study the efficient way of treatment.Methods The children with SBO were check out by X ray from a total of 121 children with bedwetting.Their parents were asked to complete the enuresis questionnaires.Urine routine test and B-ultrasound examination about kidney,bladder and ureter were also asked to be done.The clinic data of the 49 children were attained and analyzed.They were randomly divided into 2 groups,and given the controlled treatment.Group A[used 1-deamino-8-D-arginine vas-opressin(DDAVP)only] and group B(used DDAVP plus oxybutynin plus bladder training)treated for 12 weeks.Results There were totally 49 bedwetting children companied with SBO,and most of them(44 cases,89.8%)were severe type(bedwetting times≥7 times/week).Some of them coexisted with frequency,urgency,gentle urgency incontinence and microscopic hematuria(22 cases).Thirty cases were found the functional bladder capacity(FBC)decrease by B-ultrasound.The cure rates were 58.3%(group A)and 88.0%(group B)respectively.The relapse rates were 36.8%(group A)and 12.5%(group B)respectively after stopping treatment for 3 months.Conclusions SBO accounts for considerably higher rate in enuretic children.It might cause the disability of bladder function.The treatment plan with DDAVP plus oxybutynin plus bladder training can not only increase the cure rate but also lower the relapse rate.
7.Study on the homology of imipenem-resistant Acinetobacter baumannii and the genotype of carbapenemase
Xiao-Xing DU ; Xing-Guo ZHANG ; Hua ZHOU ; Yun-Song YU ; Ya-Gang CHEN ; Lan-Juan LI ;
Chinese Journal of Infection and Chemotherapy 2006;0(04):-
Objective To characterize the antibiotic resistance,homology and carbapenemase genotypes of imipenem resistant Acinetobac1ter baumannii isolated from our hospital,and analyze the clonal relatedness of the test strains.Methods Ninety five strains of imipenem resistant A.baumannii were isolated from August 2003 to December 2004 in the First Affiliated Hospital, College of Medicine,Zhejiang University.The MICs of 16 antimicrobial agents against these strains were determined by agar dilution and E-test method.The homology of these isolates was analyzed by pulse-field gel electrophoresis(PFGE).The coding gene of carbapenemases was amplified.PCR products were purified,cloned and sequenced.Plasmid DNA was extracted and purified.Conjugation and Southern blot were performed to locate the position of oxa 23 gene.Results The resistance rates to ampicillin-sulbactam and cefoperazone sulhactam were 67.9% and 30.2%.Polymyxin E had the lowest resistance rate of 17%. The resistance rate to other antimicrobial agents was higher than 90%.The 95 strains,isolated from 10 clinical units,were classified into 6 clones.Clones A and B were predominant clones.All strains produced carbapenemases which were confirmed as OXA 23 by PCR and sequencing analysis.No plasmid was extracted and conjugation was not successful.Southern bolt showed that oxa-23 gene was located on Apal-digested chromosomal segments about 220 kb and 200 kb in Clones A and B,re spectively.Conclusions OXA 23-producing A.baumannii has become one of the most important multi-resistant pathogens in our hospital.Clones A and B have widely spread in our hospital.Oxa-23 gene is located on chromosomal DNA.
8.Mechanism of BCL2L2-PABPN1 expression induced by sodium arsenite and its metabolites in 16HBE cells
SHI Ya YIN Jin yao WU Jiang JIANG Cheng lan ZHAO Rui huan ZHOU Qian HE Yue feng
China Occupational Medicine 2022;49(05):522-
Objective - - (BCL2L2)- ( )
To investigate the differential expression of the fusion gene BCL 2 like protein 2 poly A
(PABPN1) ( )
binding protein nuclear 1 induced by sodium arsenite SA and its methylated metabolites in 16HBE cells and the
Methods ) ,
related mechanism. i The 16HBE cells exposed to SA at concentrations of 1.5 3.0 and 4.5 µmol/L were set as
-, - -
low medium and high dose arsenic exposure groups. The 16HBE cells exposed to 4.5 µmol/L monomethylarsonic acid
( ), ( ) ,
MMA dimethylarsonic acid DMA and SA were set as MMA group DMA group and SA group. The 16HBE cells without
, BCL2L2-PABPN1
toxic stimulation were set as control group. After the cells were cultured for 48 hours the expression of was
- ( - ) ) ( )
detected by quantitative real time polymerase chain reaction qRT PCR . ii Two small interfering RNA siRNA silencing
基金项目:国家自然科学基金( ); 年云南省科技厅昆明医科大学应用基础研究联合专项面上项目
82160607 2021
( )
202101AY070001-054
作者简介:施雅( —),女,在读大学本科生,主要从事劳动卫生与环境卫生学研究;尹锦瑶( —),女,在读劳动卫生与环境卫
2001 1995
生学硕士研究生,主要从事劳动卫生与环境卫生学研究;施雅和尹锦瑶为共同第一作者
通讯作者:何越峰教授,博士研究生导师,- :
E mail heyuefeng@kmmu.edu.cn中国职业医学 年 月第 卷第 期 , , , · ·
2022 10 49 5 Chin Occup Med October 2022 Vol.49 No.5 523
BCL2L2-PABPN1, -
fragments were designed and transfected into 16HBE cells to knockdown which were set as siRNA 1 group
- - BCL2L2-PABPN1
and siRNA 2 group. Non transfected control group without knockdown of transfection was set up. After
, BCL2L2-PABPN1 -
culturing for 48 hours the expression level of in the three groups of cells was detected by qRT PCR. The cell
-
survival rate and early apoptosis rate were detected by MTS method and JC 1 mitochondrial membrane potential detection
, ( ) ,
method respectively. The apoptosis was detected by Hoechest33342/propidium iodide PI double staining and the expression
- Results )
level of P53 signaling pathway related proteins was detected by Western blotting. i The relative expression of
BCL2L2-PABPN1 (P ) BCL2L2-
in 16HBE cells increased with the increasing SA doses <0.01 . The relative expression of
PABPN1 - , - -
in high dose arsenic exposure was higher than that in control group low dose and medium dose arsenic exposure
( P ) BCL2L2-PABPN1 ,
groups all <0.05 . The relative expression of in SA group was higher than those in control group MMA
( P ) BCL2L2-PABPN1
group and DMA group all <0.05 . The relative expression of showed no significant difference between
, ( P ) ) BCL2L2-PABPN1
control group MMA group and DMA group all >0.05 . ii The relative expression levels of and cell
- - - ( P )
survival rate in siRNA 1 group and siRNA 2 group were lower than those in non transfected control group all <0.05 .
, (P )
However there was no significant difference in the early apoptosis rate among the three groups >0.05 . The results of
-
Hoechest33342/PI double staining showed that the number of nuclear shrinkage and early apoptotic cells in siRNA 1 group and
- - , -
siRNA 2 group was higher than that in non transfected control group. The relative protein expression levels of P53 phospho
, - - , - - ( P )
p53 BCL 2 associated death promoter P21 and cytochrome C in siRNA 1 group and siRNA 2 group were higher all <0.05 ,
- - P
and the relative protein expression levels of P53 up regulated modulator of apoptosis were lower (all <0.05), when compared
- Conclusion
with the non transfected control group. SA may block the apoptosis of 16HBE cells by inducing the expression of
BCL2L2-PABPN1
fusion gene . The mechanism may be related to the activation of P53 signaling pathway. The SA methylated
BCL2L2-PABPN1 BCL2L2-PABPN1 -
metabolites MMD and DMA had no effect on the expression of . may affect anti apoptosis
BCL2L2 PABPN1
through affecting the synergistic effect of and genes.
9.Correlation between myocardial ischemia and carotid atherosclerosis in hypertensive patients.
Tian-hui YOU ; Ya-qin LU ; Zuo-jun TIAN ; Yu-lan ZHOU ; Tao WANG
Journal of Southern Medical University 2010;30(2):311-314
OBJECTIVETo analyze the correlation between myocardial ischemia and carotid atherosclerosis in hypertensive patients.
METHODSThe clinical data were collected from 85 hospitalized hypertensive patients admitted between May 2005 and September 2008 without the complication of coronary artery disease as confirmed by cardiac computed tomographic angiography (CTA). According to the results of treadmill exercise test, the patients were divided myocardial ischemia group and ischemia-free group. Univariate and multivariate analyses were used to screen the risk factors of myocardial ischemia. The correlations were analyzed between myocardial ischemia, common carotid artery intima-media thickness (IMT), Crouse score of the carotid plaque, thickness of the intraventricular septum and left artrium. The receiver operating characteristic (ROC) curves were used to evaluate the sensitivity and specificity of IMT and Crouse score in predicting the presence of myocardial ischemia in hypertensive patients.
RESULTSCarotid plaque formation was identified as the major risk factor of myocardial ischemia in hypertensive patients (OR=4.982, P=0.004). The incidence of myocardial ischemia in the hypertensive patients with carotid plaques was significantly higher than that in the patients without the plaque (Chi2=9.317, P=0.002). Myocardial ischemia in hypertensive patients was positively correlated to the thickness of the intraventricular septum (r=0.362, P=0.001) and left artrium (r=0.298, P=0.009), and the IMT of the common carotid artery was positively correlated to the thickness of the intraventricular septum (r=0.231, P=0.045). The area under cure (AUC) of the ROC curve of Crouse score was 0.726-/+0.061 in predicting the presence of myocardial ischemia in the hypertensive patients (P=0.001), and that of IMT was 0.682-/+0.061 (P=0.006).
CONCLUSIONCarotid plaque formation is the major risk factor of myocardial ischemia in hypertensive patients and shows a positive correlation to the onset of myocardial ischemia, but both the common carotid artery IMT and the Crouse score of the carotid plaque are not accurate markers for predicting myocardial ischemia in patients with hypertension.
Adult ; Aged ; Aged, 80 and over ; Atherosclerosis ; complications ; pathology ; Carotid Artery, Common ; diagnostic imaging ; pathology ; Coronary Angiography ; methods ; Female ; Humans ; Hypertension ; complications ; pathology ; Male ; Middle Aged ; Multivariate Analysis ; Myocardial Ischemia ; etiology ; pathology ; Risk Factors ; Tomography, X-Ray Computed ; Ultrasonography
10.Application of membrane protein-based two-dimensional electrophoresis in chondrocyte- related investigations.
Jun XIAO ; Zhan-jun SHI ; Zhan-dong ZHAO ; Ya-peng ZHOU ; Tian LAN
Journal of Southern Medical University 2010;30(12):2705-2707
OBJECTIVETo explore the feasibility of membrane protein-based two-dimensional electrophoresis (2-DE) in the investigations of chondrocyte-related diseases and its complementarity with total protein-based 2-DE.
METHODSKnee cartilage samples were obtained to isolate the chondrocytes with type II collagenase/hyaluronidase digestion. The membrane proteins and total proteins were extracted and loaded separately onto PH3.0-10.0 non-linear gel strip for 2-DE analysis. The qualities of membrane protein-based 2-DE gels were evaluated, and the difference between the distribution profiles of the membrane protein spots and that of the total protein were observed and their complementarities were evaluated.
RESULTSMembrane protein-based 2-DE generated high-quality gel images, and on each gel 412.3±13.5 protein spots were identified. These spots were distributed in the region of isoelectric point pH 5.0-9.0. In the gel images generated by the total proteins, 564.3±5.9 protein spots were identified in each image, and the spots were distributed in the region of isoelectric point pH 3.0-7.0.
CONCLUSIONMembrane protein-based 2-DE of the chondrocytes can generate high-quality gel images, and the isoelectric distribution of the protein spots is complementary to that of total protein, which provides valuable information for chondrocyte-related diseases.
Adult ; Chondrocytes ; chemistry ; Electrophoresis, Gel, Two-Dimensional ; methods ; Female ; Humans ; Isoelectric Point ; Male ; Membrane Proteins ; isolation & purification ; Middle Aged