1.Effect of high glucose on the expression of KLF6 in human lens epithelial cell
Hong-tao, WANG ; Yong-zhen, BAO
Chinese Journal of Experimental Ophthalmology 2012;30(2):136-140
Background Epithelial-mesenchymaltransition (EMT)isamajorcontributortothe pathogenesis of posterior capsular opacification(PCO).Kruppel-like factor 6 (KLF6) is a zinc finger protein,which can be stimulated by high glucose in proximal tubule cells and involved in transforming growth factor beta (TGF-β)induced EMT of diabetic nephropathy.ObjectiveThis study was designed to investigate the effect of high glucose on the expression of KLF6 and its target genes( TGFB1,TGFBR1,COLIA1,HSP47) in human lens epithelial cells (LECs).MethodsHuman LECs(SRA01/04) were cultured and exposed to different concentration of glucose.The expressions of KLF6 mRNA and protein were analyzed by real time polymerase chain reaction( real time PCR) and western blot after treatment with high glucose.The expressions of KLF6 target genes were analyzed by real time PCR to evaluate the EMT of SRA01/04 cells.ResultsCompared with the control group(5.5 mmol/L),the relative mRNA levels of t-KLF6 and wt-KLF6 in SRA01/04 treated with high glucose(22.2,44.4,66.6 mmol/L) increased obviously (F =72.53,42.02,P<0.01 ).Then,the concentration of 22.2 mmol/L was used in the next experiments.The relative mRNA levels of t-KLF6 and wt-KLF6 increased to the peaks after treatment with high glucose for 12 h,and began to decrease after 24 h until lower levels after 48 h ( F =100.12,125.52,P < 0.01 ).Western blot showed that the expression of KLF6 protein was also upregulated by high glucose treatment.With the promotion of the expression of KLF6 gene,the relative mRNA levels of TGFB1,TGFBR1,COLlAl and HSP47 of treated cells also respectively increased after treatment for 12 h,and began to decrease after 24 h until nearly at the levels of the control groups after 48 h( F=6.73,162.35,64.39,12.05,P<0.05 ).ConclusionsIt was concluded that high glucose induced the expression of KLF6 in human LECs,and KLF6 transiently stimulated the expression of target genes TGFB1,TGFBRl,COLlAl and HSP47 which were mainly involved in the mechanism of EMT.
2.Protection of quercetin against hyperbaric oxygen-induced apoptosis of human lens epithelial cell and its mechanism
Li-bin, CHANG ; Yong-zhen, BAO ; Yi, CHEN ; Wen-zhen, YU ; Xiao-xin, LI
Chinese Journal of Experimental Ophthalmology 2012;30(6):485-489
Background Oxidative stress-induced apoptosis of human lens epithelial cells (LECs) is associated with c-Jun N terminal kinase (JNK) pathway.Quercetin possesses the antioxidation by inhibiting the JNK pathway.However,whether quercetin can protect LECs from the oxygen-induced damage is still not proved.Objective This study attempted to invatigate the effects and its mechanism of quercetin against hyperbaric oxygeninduced LECs apoptosis. Methods Human LECs line SRA01/04 was cultivated and passaged in MEM medium containing 10% fetal bovine serum and 0.5% non-essential amino acids for 2 hours,with or without 20 μmol/LSP600125 or 1 μmol/L quercetin prior to exposure to hyperbaric oxygen.Each exposure session remained 6 hours in 99% O2 and 1%CO2 with a pressure chamber at 588 kPa.The viability of human LECs was detected by MTT.Cell apoptosis was assessed by flow cytometer using Annexin V-FITC apoptosis detection.The expression of JNK/p-JNK,c-Jun/p-c-Jun,caspase 3 and caspase 9 were detected by Western blot. Results LECs viability (A570 ) was 0.835 ±0.082,0.450±0.083,0.654±0.079,0.649±0.090 respectively in the blank control group,hyperbaric oxygen exposed group,hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group.The A570 in the hyperbaric oxygen exposed group was significantly lower than the blank control group ( P =0.000),but those in hyperbaric oxygen + SP600125 group and hyperbaric oxygen+quercetin group were significantly higher than the hyperbaric oxygen exposed group ( P =0.003,0.002 ).The numbers of apoptosis cells were 3.17 ±0.74,19.77 ± 1.44,8.45 ±0.93,7.79 ±0.78 respectively in the blank control group,hyperbaric oxygen exposed group,hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group.Apoptotic LECs were significantly increased in the hyperbaric oxygen exposed group compared with the blank control group ( P=0.000),but those in the hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group were significantly reduced in comparison with hyperbaric oxygen exposed group (both P=0.000).In additional,expressions of p-JNK,p-c-Jun,caspase 3 and caspase 9 proteins in the cells were elevated in the hyperbaric oxygen exposed group compared with the blank control group (all P =0.000 ),however,those in the hyperbaric oxygen + SP600125 group and hyperbaric oxygen + quercetin group were declined when compared with the hyperbaric oxygen exposed group( all P<0.05 ). Conclusions JNK pathway is involved in the apoptotic procedure of human LECs induced by oxygen stress.SP600125 and certain concentration of quercetin can interdict the JNK signal pathway and endogenous apoptosis of LECs and further alleviate hyperbaric oxygen-induced damage of LECs.
4.Effects of ultraviolet-B laser irradiation on lens α-crystallin and protection of indole-3-carbinol to chaperone activity of α-crystallin
Qin, ZHANG ; Xiao-guang, CAO ; Xue-ting, PEI ; Yong-zhen, BAO
Chinese Journal of Experimental Ophthalmology 2012;30(6):538-542
Background Ultraviolet radiation is one of factors of the formation of age-related cataract.Indole-3-carbinol(I3C) is a plant chemical material with inhibitory effect on oxidative-induced cell damage and formation of amyloid fibrils,and the oxidative damage and amyloid fibrils are associated with cataract.However,the relationship between I3C and α-crystallin is in study. Objective This study was to evaluate the effects of ultraviolet-B laser irradiation on the secondary structure of α-crystallin and to explore the protection of I3C to chaperone activity of α-crystallin. Methods The fresh eyeballs were obtained from 1-year-old cattle to prepare the purified lens α-crystallin by gel chromatography.α-Crystallin was isolated from cattle lenses using gel chromatography.The purified α-crystallin was collected using fast protein liquid chromatography ( FPLC ) and exposed to 1:308 nmultraviolet-B at different irradiation intensities ( 23.75,118.75,475.00,1187.50,2375.00,4750.00,11 875.00,23 750.00 mJ/cm2 ) and then to ultraviolet-B 2:308 nm with irradiation intensities of 28 535.00,6730.00,3435.00,1910.00,1040.00 mJ/cm2.Ultraviolet-absorbance spectra,tryptophan fluorescence and N-formylkynurenine (N-FK)fluorescence spectra of both irradiated and non-irradiated α-crystallin were measured.I3C at the concentrations of 50 μmol/L and 100 μmoL/L were added to the α-crystallin solution to perform a catalase (CAT) thermal aggregation to confirm the chaperone activity of the α-crystallin,and the α-crystallin solution without any I3C was used as control.The ratios of A360 between various intervene groups with control group were calculated using spectrophotometry.Results The A280 values of the α-crystallin declined to 10% at the ultraviolet-B irradiation intensity of 1187.5 mJ/cm2 and that at the intensity of 23.75 J/cm2 lowed to 2%.A negative correlation was seen between the ultraviolet-B irradiation intensity and the A280 value of the α-crystallin (R2=0.925 ) and a positive correlation was found between ultraviolet-B with N-FK ( R2 =0.949 ).Ultraviolet-B irradiation intensity showed a negative correlation with Trp fluorescence intensity (R2 =0.996 ).CAT hot condensed experiment revealed that after addition of different concentrations of indole-3-carbinol,the relative A360 values at various ultraviolet-B irradiation group were significantly higher than those of the control group (P =0.000),and the decreasing degree of chaperone activity of α-crystallin was lower than that of the control group ( P =0.000 ). Conclusions The study suggests that I3C can protect the chaperone activity of α-crystallin from photooxidation,and the ultraviolet-B laser may be a good exposure source compared with ultraviolet lamp.The ultraviolet-B laser irradiation causes the alteration of structure and chaperone activity of α-crystallin.
5.Anterior cervical Cage-assisted fusion combined with locking titanium plate internal fixation for the treatment of extension-type cervical fracture
Jian WU ; Yanxi LIU ; Xingxing QIN ; Yong ZHENG ; Zhen SHI ; Tongzhu BAO
Chinese Journal of Tissue Engineering Research 2013;(43):7546-7551
BACKGROUND:There is no literature about the treatment according to the mechanism of cervical spine injury classification, especial y for the treatment of extension type cervical fracture/dislocation with merger cases of posterior composite structure damage, whether simple anterior approach can meet the needs of the treatment has no detailed elaboration. This article may analyze from the aspect of cervical spine injury mechanism. OBJECTIVE:To observe the short-term effect of anterior cervical Cage-assisted fusion combined with locking titanium plate internal fixation for the treatment of extension type cervical fracture. METHODS:A retrospective analysis was performed in 15 extension type cervical spine fracture dislocation patients treated with decompression anterior cervical intervertebral disc resection plus bone graft with cage-fusion locking titanium plate internal fixation from June 2006 to March 2011 in the Department of Orthopedics, Xianning Central Hospital, including 10 cases of single segment injury and treatment, and five cases of multiple segment injury and treatment. Japanese Orthopaedic Association score and the neck disability index were compared before and after treatment;the cervical flexion and height were measured according to the antersposterior X-ray film taken before fixation, 1 week after fixation and final fol ow-up.RESULTS AND CONCLUSION:The patients were fol owed-up for 8-37 months. One case had Cage mild sinking and shift, and there was no internal fixation breakage or loosening in al the patients. Transient pharyngeal discomfort was observed in 11 patients. Compared with the preoperation, the Japanese Orthopaedic Association score, neck dysfunction index, fusion segment cervical flexion and fusion segment intervertebral disc height were significantly improved at 1 week after fixation and final fol ow-up (P<0.05). There were no significant differences between 1 week after fixation and final fol ow-up (P>0.05). The short-term effect of decompression anterior cervical intervertebral disc resection plus bone graft with cage-fusion locking titanium plate internal fixation for the treatment of extension-type cervical fracture is good.
6.Analysis of the Etiology of Pulmonary Arterial Hypertension in 276 Children
xue-qin, LIU ; jun-bao, DU ; yong-hong, CHEN ; yu-wen, MA ; wan-zhen, LI
Journal of Applied Clinical Pediatrics 2006;0(13):-
Objective To investigate the etiology and clinical characteristics of pediatric pulmonary arterial hypertension(PAH) and improve its early diagnosis and treatment.Methods The clinical and echocardiogram data of all inpatients with PAH in Pediatric Department of Peking University First Hospital between May 1995 and May 2007 were retrospectively analyzed for age,sex,etiology,symptoms and echocardiographic measurement of pulmonary artery pressure.Data were divided into groups according to different etiology and statistics.Pulmonary arterial systolic pressure(sPAP) values estimated from the tricuspid regurgitant velocity by Doppler echocardiography were compared among different groups.Cases who were not belonged to the first category of the Venice Clinical Classification of pulmonary hypertension were not included.Results Totally 276 cases,168 boys and 108 girls were diagnosed to have PAH.Age ranged from 1 month to 17 years,median age was 9 months.Most of pediatric PAH was associated-PAH(267 cases,96.7%),while idiopathic PAH took a small part(9 cases,3.3%).Congenital heart disease-associated PAH(CHD-PAH) was predominant(245 cases,88.7%) and left to right shunt was the main lesion (217 cases,88.6%),while complex lesion-associated PAH comprised 28 cases(11.4%).Connective tissue disease associated PAH(CTD-PAH) was the second common among this group of pediatric PAH patients(19 cases,6.9 %).The incidence of PAH in systemic lupus erythematosus(SLE),juvenile rheumatoid arteritis and takayasu arteritis were 10.3 %(13/126),8.7%(4/46),15.4%(2/13),respectively.The other 3 cases of PAH were associated with portal hypertension(2 cases) and thalassanemia(1 case).The estimated sPAP from tricuspid regurgitant velocity in 8 cases with idiopathic PAH[(74.6?23.9) mmHg(1 mmHg=0.133 kPa)]was higher significantly compared with those of 33 cases of CHD-PAH [(58.0?19.7) mmHg ] and 12 cases of CTD-PAH [(49.6?18.9) mmHg] respectively(t=-2.052,-2.609 Pa
7.Etiologic and clinical characteristics of syncope in children.
Qing-you ZHANG ; Jun-bao DU ; Jiong QIN ; Yong-hong CHEN ; Wan-zhen LI ; Xin-hua BAO
Chinese Journal of Pediatrics 2007;45(1):59-63
OBJECTIVESyncope is a common problem in children and adolescents. Such an event may have multiple possible causes, ranging from benign conditions to life-threatening diseases. Syncope is a major challenge for the practicing physicians. It is very important to know the etiologic and clinical characteristics of syncope in children. This study aimed to improve diagnostic efficacy of syncope in children by analyzing the etiology and clinical characteristics of syncope.
METHODSThe investigators retrospectively analyzed the causes of syncope and diagnostic workup of 154 consecutive children seen in Department of Pediatrics, Peking University First Hospital because of a syncopal event.
RESULTSAutonomic-mediated reflex syncope (AMS) was the most common cause of syncope (65.6%), whereas cardiac disorders were found in 10 cases (6.5%) comprising the second cause of syncope in children. Other causes included psychologic problems and neurological and metabolic disorders. Although many causes were studied, 25 cases (16.2%) were found to have uncertain etiologies yet. The children with AMS were commonly seen in pubertal girls, and they had clear inducement of syncope and prodromes. The children with cardiac syncope often had history of cardiac diseases, and they were often younger than those with AMS. Lack of prodromes of syncope, exercise-related syncope, syncope spells seen in any body position, frequent syncope spells and sudden death in family were clues of cardiac syncope. Neurological disorders should be considered if there are any of the followings: syncope with seizure activity, syncope spells seen in any position, and a postictal phase of disorientation or neurologic abnormal signs. A metabolic cause was entertained when the child had a history of metabolic diseases, prolonged anger, or violent vomiting and diarrhea. Children with psychiatric disorders were adolescent girls with prolonged syncope spells, and had more frequent syncopal episodes. Most children with syncope were evaluated by many of diagnostic tests, but most of those tests were not goal-directed approach. Since persons with cardiac syncope were at increased risk for death from any cause, electrocardiography was recommended in almost all children with syncope. Neurologic testing including electroencephalography, computed tomography, etc. were rarely helpful unless neurologic signs and symptoms are present. Holter electrocardiography and echocardiography were most useful in children with suspected cardiac syncope. There was little benefit of screening cardiac enzyme in children with syncope. Routine blood tests (blood electrolytes and blood glucose, etc) rarely yield diagnostically useful information unless the children had the history of metabolic diseases. Head-up tilt testing was most useful in children with recurrent syncope in whom heart disease was not suspected. The children with frequent syncope, long lasting syncopal episode and clear psychiatric inducement of syncope should be evaluated by psychiatric testing.
CONCLUSIONSyncope in children may result from a wide variety of causes, and clinicians often use a wide range of investigation to try to achieve a diagnosis. But most of investigations have low diagnostic yield. Thorough history taking, physical examination and electrocardiography are the core of the syncope workup.
Adolescent ; Child ; Child, Preschool ; Electrocardiography ; Female ; Humans ; Male ; Retrospective Studies ; Syncope ; diagnosis ; etiology
8.NR2B-pERK1/2-pElk-1 signaling contributes to the avoidance learning and memory of rats.
Xu-Hong CHEN ; Si-Yun SHU ; Zhen-Jiang LIANG ; Xin-Min BAO ; Li-Xue CHEN ; Yong-Ming WU
Chinese Journal of Applied Physiology 2007;23(1):121-125
AIMTo investigate whether NR2B-pERK1/2-pElk-1 signaling contributes to the Y-maze learning and memory of rat brain.
METHODS45 adult male SD rats were divided into 4 groups: (1) Ifenprodil peritoneal injection group (Ifenprodil ip, n = 14); (2) DMSO peritoneal injection group(DMSO ip, n = 15); (3) Ifenprodil cerebral ventricle injection group (Ifenprodil ic, n = 8); (4) DMSO cerebral ventricle injection group(DMSO ic, n = 8). Y-maze training and test were used as an learning and memory enhancing stimulus. Immunohistochemical and Western blotting methods were used for detecting pERK1/2 and pElk-1 expression intensity of different brain regions.
RESULTSCompared with the DMSO ip group, the ifenprodil ip group showed no change on the Y-maze learning score (P > 0.05), but its Y-maze memory score tested 24 after learning decreased (P < 0.05). Ifenprodil peritoneal injection made brain pERK1/2 and pElk-1 expression decreased generally. In hippocampus, marginal division of striatum(MrD), amygdala,these changes were more significant (P < 0.05). Compared with the DMSO ic group, the reconsolidation of Y-maze memory tested 6 hours after ifenprodil injection was impaired in ifenprodil ic group (P < 0.05). The OD value of pERK1/2 and pElk-1 positive bands in ifenprodil ic group attenuated generally. The pElk-1 positive bands of caudate putamen and MrD almost disappeared in ifenprodil ic group.
CONCLUSIONNR2B is essential for the formation of long-term memory, reconsolidation of Y-maze memory. The deactivation of NR2B by ifenprodil will impair these courses. Meanwhile, the deactivation of NR2B attenuates pERK1/2 and pElk-1 expression of learning and memory related regions after Y-maze learning and memory reconsolidation test. In MrD and caudate putamen, the pElk-1 expression are completely blocked by ifenprodil after memory reconsolidation test.
Animals ; Avoidance Learning ; physiology ; Dimethyl Sulfoxide ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Male ; Maze Learning ; physiology ; Memory ; physiology ; Piperidines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; ets-Domain Protein Elk-1 ; metabolism
9.Application of the nasal lining flap to correct the wide unilateral complete cleft lip.
Ai-qun LI ; Bao-chun LIU ; Yong-gang SUN ; Zhen-kang ZHANG ; Guang-he WANG
Chinese Journal of Stomatology 2003;38(2):135-136
OBJECTIVETo study use of nasal lining flap in correction of the wide complete unilateral cleft lip.
METHODS78 patients with wide complete cleft lip were. operated, firstly to be sure. That the lip length on both sides was equal. Then the nasal lining flap was used to enhance the height of the lip on cleft side for keeping symmetry with the non-cleft side. The rest of procedures was the same as "Millard" method.
RESULTS84.6 percent of patients had good appearance at one and half years after surgery.
CONCLUSIONSIt is an effective way to take advantage of the nasal lining flap to correct the wide complete unilateral cleft lip.
Cleft Lip ; surgery ; Humans ; Infant ; Male ; Nose ; surgery ; Reconstructive Surgical Procedures ; Surgical Flaps
10.Prediction and analysis model of temperature and its application to a natural ventilation multi-span plastic greenhouse equipped with insect-proof screen.
Shu-zhen LIU ; Yong HE ; Yu-bao ZHANG ; Xiang-wen MIAO
Journal of Zhejiang University. Science. B 2005;6(6):523-529
The natural ventilation widely used in greenhouses has advantages of saving energy and reducing expense. In order to provide information for climate control of greenhouse, a model was developed to predict the variation of air temperature in the naturally ventilated greenhouse equipped with insect-proof screen. Roof ventilation and combined roof and sidewall ventilation were considered in the model. This model was validated against the results of experiments conducted in the greenhouse when the wind was parallel to the gutters. The model parameters were determined by the least squares method. In the used model, effects of wind speed and window opening height on the air temperature variation were analyzed. Comparison between two types of ventilation showed that there existed a necessary ventilation rate which results in air temperature decrease in natural ventilation under special climatic conditions. In our experiments when wind speed was less than 3.2 ms(-1), wind had a more gradual effect on greenhouse temperature for roof ventilation, compared with combined roof and sidewall ventilation, which had greater air temperature decrease than roof ventilation only.
Air Conditioning
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instrumentation
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methods
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Air Movements
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Computer Simulation
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Computer-Aided Design
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Equipment Design
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Equipment Failure Analysis
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methods
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Insect Control
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instrumentation
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methods
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Models, Theoretical
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Plant Development
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Plastics
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Rheology
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instrumentation
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methods
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Temperature
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Wind