2.Effect of suppression of platelet-derived growth factor-α receptor expression with antisense oligonucleotide on proliferation and apoptosis of retinal pigment epithelium cell
Yan-yi, PENG ; Mei-yuan, QIU ; Zhi-xiang, DING ; Miao-yun, LIAO ; Cai-wen, FAN
Chinese Journal of Experimental Ophthalmology 2012;30(4):341-345
BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10% fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.
3.Morphologic Localization and Purified Cultivation of Sinoatrial Nodes Isolated from Newborn Rabbits
qing-zhi, WANG ; guang-lan, MAO ; lei, YU ; xiao-zhong, QIU ; lin, YUAN
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To localize the sinoatrial node (SAN) of the newborn rabbits in vivo and cut it for purifying cultivation and study the morophologic characters of primary cultured pacemaker cells of SAN under light microscope and transmissional electron microscope. Methods Hearts of the newborn rabbits were embedded in paraffin for HE-staining and observed the location, form of SAN under optical microscope; SAN cells isolated from neonatal rabbits cultured and purified with the method of differential attachment and BrdU-treatment.Results SAN localized in the anterior wall of the superior vena cava and the posterior-lateral atrial wall.There was about 0.32 mm between its lowest point and sulcus terminalis. Three distinctly different types of cells were observed among the cultured cells of SAN: spindle, araneiform and polygon. The spindle cells covered the greatest proportion of the cultured cells of SAN (59.6%?7.3%). The frequency of spontaneous contraction of spindle cells was the highest among the constrcting cells (145 ?9)time/min. The results of ultrastructure observation showed that myofibrils and other organelles in spindle cells were poorly organized and significantly decreased in number compared with araneiform cells. There was no significant difference between araneiform cells isolated from SAN and from atrial muscle.Conclusion Among the cultured cells from neonatal rabbits SAN, the spindle cells are the pacemaker cells of SAN.
4.Adrenal tuberculosis:contrast-enhanced CT features with clinical correlation
Zhi-Gang YANG ; Ying-Kun GUO ; Yuan LI ; Peng-Qiu MIN ; Jian-Qun YU ;
Chinese Journal of Radiology 1999;0(10):-
Objective To investigate the correlation between specific features on contrast-enhanced CT and its clinical findings of adrenal tuberculosis so as to improve the diagnostic accuracy of the disease. Methods Contrast-enhanced CT features in 30 patients with documented adrenal tuberculosis were retrospectively evaluated blindly for the features of location,size,morphology,attenuation and enhancement patterns on CT images,and compared with clinical and pathological materials.Results The common CT manifestations were as follows:enlargement of the adrenal glands in all 30 cases(bilateral 90%,mfilateral 10%)including mass-like enlargement in 13 cases and enlargement but the contours preserved in 17 cases, heterogeneity(28 cases,93.3%),calcification(17 cases,56.7%),and low attenuation in the center with peripheral enhancement(16 cases,53.3%)of the lesions.After antituberculosis chemotherapy, 5 cases of enlarged adrenal glands decreased in size or returned to normal size and configuration,with disappearance of the central low attenuation and new appearance of dot-like calcification in 2 cases.Cochran Armitage trend test showed there was an increasing tendency of calcification rate with clinical duration(X~2= 7.47,P
5.Role of NRF2 signaling pathway in trichloromethane-induced oxidative stress in L02 cells
TAN Shi ying ZHONG Yuan fang QIU Yi bing ZOU Zhi hui YU ri an
China Occupational Medicine 2022;49(05):530-
Objective - ( )
To investigate the effects of nuclear factor erythroid 2 related factor 2 NRF2 on the oxidative stress
( ) Methods ) ,,
induced by trichloromethane TCM in human normal hepatocyte L02 cells. i L02 cells were stimulated with 1 2
, , , ( ),
4 8 12 16 and 20 mmol/L TCM solution dissolved in dimethyl sulfoxide and the control group and blank group were set
, - ,
up. After culturing for 24 hours the cell viability was detected by CCK 8 colorimetric method and the concentration of TCM
) -, -
stimulation was screened. ii L02 cells in logarithmic growth phase were randomly divided into control group and low medium
- , , ,
and high dose groups. After 24 hours of exposure to 0 4 8 and 12 mmol/L TCM the cells were collected. The activity of
( ), ( ), ( - ) ( )
superoxide dismutase SOD catalase CAT glutathione peroxidase GSH Px and the level of malondialdehyde MDA
NRF2, - (HO-1),
were detected by colorimetric analysis. The mRNA expression levels of heme oxygenase 1 glutamate cysteine
(GCLC) () (NQO1) -
ligase catalytic subunit and NAD P H quinone dehydrogenase 1 were detected by real time fluorescence
, - ,
polymerase chain reaction. The protein levels of NRF2 HO 1 GCLC and NQO1 were detected by Western blotting.Results ) , , , ,
i When the concentration of TCM was 4 8 12 16 and 20 mmol/L the survival rate of L02 cells decreased
( P ) , ,
significantly compared with the control group all <0.05 . The concentration of 0 4 8 and 12 mmol/L were selected as the
) , -
stimulation doses for subsequent experiments. ii Compared with the control group the activities of SOD and GSH Px in L02
( P ) ( P ), -
cells in the three doses groups decreased all <0.05 and the levels of MAD increased all <0.05 with a dose effect
- (P ),
relationship. The CAT activity of L02 cells in the medium dose group was lower than that in the control group <0.05 and the
- ( P )
CAT activity of L02 cells in the high dose group was lower than that in the others three groups all <0.05 . Compared with the
, NRF2 - (P ),NRF2
control group the relative expression levels of mRNA in L02 cells in the low dose group decreased <0.05
- (P ), NRF2
mRNA in L02 cells in the medium dose group increased <0.05 mRNA and NRF2 protein expression in L02 cells in
( P ) HO-1,GCLC, NQO1 ,
the highdose group increased both <0.05 . The relative expression level of mRNA and GCLC NQO1
( P )
protein expression in L02 cells in the three doses groups increased compared with the control group all <0.05 . The relative
NRF2 - - -
expression level of mRNA in L02 cells in the high dose group was higher than that in the low and medium dose groups
( P ), - (P ),
both <0.05 and the relative expression of NRF2 protein was higher than that in the low dose group <0.05 but the
HO-1 GCLC - - (
relative expression levels of and mRNA and HO 1 protein level were lower than those in the medium dose group all
P )Conclusion -
<0.05 . TCM exposure can inhibit the proliferation of L02 cells by inducing oxidative stress with a dose effect
,
relationship. In this process the antioxidant mechanism mediated by NRF2 was activated. The expression of antioxidant defense
, - ,
and detoxification related target genes downstream of NRF2 signaling pathway was activated and the expression of HO 1
-
GCLC and NQO1 was up regulated to alleviate the oxidative damage caused by TCM.
6.The Advance in Research of Banana Bunchy Top Virus
Huan-Ge ZHAO ; Sheng-Niao NIU ; Yuan-Gang HUA ; Shi-Ming QIU ; Da-Xin WANG ; Zhi-Xin LIU ;
Microbiology 1992;0(05):-
Banana bunchy top virus (BBTV) is the pathogen of banana bunchy top disease (BBTD); it seriously disserves the banana production. This paper reviewed the separation and purification methods, classifying and taxonomy status of BBTV; the genome structure and function of each encode protein of the virus; and the present problems that should be further clarified.
7.Protective Effect of Panax quinquefolium 20s-proto-panaxdiolsaponins on Acute Myocardial Infarction in Dogs
Da-Yuan SUI ; Xiao-Feng YU ; Shao-Chun QU ; Zhong-Zhi LU ; Li WANG ; Man-Qiu CHEN
China Journal of Chinese Materia Medica 2001;26(6):416-419
Objective: To study the protective effects of Panax quinquefolium 20s-protopanaxdiolsaponins extracted from leaves of P.quinquefolium(PQDS)on acute myocardial infarction(AMI)in dogs. Method:The parameters of myocardial infart size , the serum CK and LDH activity, myocardial metabolism,free radicals and coronary circulation etc were determined by using the model of ligation of LAD in the anaesthetized open-chest dogs. Result:In dogs treated with PQDS(in a dosage of 10 and 20 mg*kg-1 iv infusion),the myocardial infarct size, the activity of serum CK ,LDH and the contents of serum FFA and LPO were decreased,whereas the activity of serum SOD and GSH-Px increased markedly.At the same time, myocardial blood flow was increased and coronary vascular resistance decreasedsignificantly. Conclusion:PQDS has protective effect on myocardial ischemia by modifying metabolic dysfunction of FFA, inhibiting oxygen free radicalmediated peroxidation of membrane lipids , enhancing endogenous antioxidase activity and increasing myocardial blood supply.
8.Changes of the mitochondrial DNA copy number and the antioxidant system in the PBMC of hepatocellular carcinoma.
Yuan GAO ; Hong-jing NIE ; Dong YANG ; Cheng-shi DING ; Min JIN ; Zhi-qiang CHEN ; Zhi-gang QIU ; Xuan GUO ; Zhao-lila CHEN ; Jun-wen LI
Chinese Journal of Applied Physiology 2016;32(1):1-5
OBJECTIVETo investigate the relationship between the changes of the copy numbers of mtDNA in peripheral blood mono-nucle- ar cell(PBMC) and the disordered of antioxidant capacity of hepatocellular carcinoma (HCC) patients.
METHODSThe Ficoll Hypaque method was used to isolate the PBMC from blood specimens. The ND1 gene of the mitochondrial was amplified by real-time PCR; meantime β-actin was served as a quantitative standard marker; the difference of mtDNA copy number in PBMC was compared between HCC and healthy control group. The level of reactive oxygen species (ROS) in PBMC was determined by flow cytometry. The change of total antioxidant capacity (T- AOC) of plasma was detected by the biochemistry examination.
RESULTSThe copy numbers of ND1 gene in PBMC of HCC was 73% that of the healthy control group,which suggested a decrease of the copy numbers of mtDNA in HCC. The levels of ROS of PBMC in HCC was (417. 82 ± 110.62) and (301.82 ± 75.54) in control group, which showed that the levels of ROS of PBMC in HCC were significant higher than that in control group (P < 0.01).Plasma T-AOC in HCC was (1.30 ± 0.85), and (3.20 ± 1.62) in control. The T-AOC of plasma of HCC was significantly lower than in control group (P < 0.01).
CONCLUSIONThere was a certain relationship between the decrease of the copy numbers of mtDNA and the disordered antioxidant capacity in hepatocellular carcinoma, which may be associated with the development of hepatocellular carcinoma.
Actins ; Antioxidants ; metabolism ; Carcinoma, Hepatocellular ; blood ; genetics ; Case-Control Studies ; DNA Copy Number Variations ; DNA, Mitochondrial ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Liver Neoplasms ; blood ; genetics ; Reactive Oxygen Species ; metabolism
9.Clinical analysis of massive hemispheric infarction treated with moderate hypothermia.
Jian SU ; Yong-ming QIU ; Zhi-hua CHEN ; Yuan-yuan TANG ; Yan CHEN
Chinese Journal of Traumatology 2003;6(5):318-320
The potential of hypothermia in reducing neuronal damage has been demonstrated in several animal models of focal cerebral ischemia. The feasibility and safety of this technique for acute stroke has been only examined in one study, describing a total of 25 patients. We analyzed the data from 50 consecutive patients with acute stroke treated with moderate hypothermia in neurocritical care units of 4 university clinics to evaluate the feasibility and safety of moderate hypothermia.
Acute Disease
;
Adult
;
Aged
;
Cerebral Infarction
;
therapy
;
Female
;
Glasgow Coma Scale
;
Humans
;
Hypothermia, Induced
;
Male
;
Middle Aged
;
Treatment Outcome
10.Nursing and the effects of tongue surgical treatment on patients with sleep apnea syndrome
Hong SU ; Zhi-Jie LI ; Jing-Qiu ZHANG ; Hai-Ying SU ; Yuan-Yuan SHAN ; Ming-Li MENG
Chinese Journal of Modern Nursing 2012;18(7):775-777
Objective To explore the effects of tongue surgery such as ablation of the tongue,tongue traction,hyoid suspension on the patients with sleep apnea syndrome and its key nursing.Methods Totals of forty-one patients with sleep apnea syndrome received uvulopalatopharyngoplasty surgery ( UPPP ) and hard palate surgery were received preoperative psychological nursing and prepared preoperative preparation.Enhanced the observation after surgery,and payed more attention to functional training and post-operative health guidance.and the complications,blood pressure and therapy effects were observed.Results Forty-one patients had well recover.Three months later,18 patients' respiratory disturbance index was ( 18.4 ± 17.3 ) significantly lower than(56.2 ± 19.3),and oxygen saturation was ( 81.9 ± 7.01 ) % significantly higher than (71.9 ± 12.1)% ( P<0.01 ).Conclusions The surgical treatment of tongue hypertrophy of the tongue and the tongue caused by tongue fall zone blocking is effective for patients with OSAHS,and perioperative nursing and postoperative functional training is very important for successful operation.