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.

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.

Adolescent ; Female ; Humans ; Mucocutaneous Lymph Node Syndrome ; complications ; Uveitis, Anterior ; etiology

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.

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.

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

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

OBJECTIVETo study the segmentation methods of the liver CT images and the value of 3-dimensional (3D) reconstruction of the liver in the planning of hepatic surgery.

METHODSThe 2D Digital Imaging and Communications in Medicine (DICOM) format data of the liver obtained from healthy volunteers were transformed into bmp format image, and the liver image segmentation was performed using Photoshop software. The 3D model was reconstructed using MIMICS software.

RESULTSThe DICOM format data of the liver obtained by 64 slice spiral CT included totally 658 slice images. The segmented liver image showed clear profiles and complete intrahepatic duct data were reserved. The segmented liver images were free of discontinuation during continuous observation. The liver surface and internal ductal system, including the hepatic arteries and veins, and the hepatic portal system and their branches, were represented clearly. The reconstructed liver allowed clear identification of the anatomic landmark and matched the actual liver volume. The reconstructed ductal structure were distinct and continuous with natural texture. The reconstructed liver and the hepatic internal duct system were simultaneously displayed by adjusting the transparency of the liver, and the blood vessels were also represented.

CONCLUSIONSegmentation of the liver images in different phases using Photoshop can be feasible for liver reconstruction. The reconstructed liver and the intrahepatic ductal structure allow vivid 3D observation of the spatial relationship among the major tracts and accurate estimation of the liver volume.

Adult ; Female ; Humans ; Image Interpretation, Computer-Assisted ; methods ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; methods ; Liver ; diagnostic imaging ; Tomography, Spiral Computed ; methods

OBJECTIVETo observe the histopathologic characteristics of 2 micron continuous wave laser transurethral partial cystectomy for the treatment of bladder tumor.

METHODSA total of 54 patients with 65 bladder tumors underwent 2 micron laser via transurethral by caudal or surface anesthesia from October 2007 to December 2008. It included 41 male and 13 female cases, and the age ranged from 27 to 81 years old with a mean of (66.2 +/- 12.4) years old. The operation evaporated and exsected the wall of urinary bladder, including tumor, submucosa and all muscular layers. Specimens were sent for pathology examination. The histomorphologic changes of raw surfaces were observed 1 week, 1 month, 3 months, 6 months and 1 year postoperation by cystoscopic and pathologic examinations.

RESULTSAll the patients tolerated in the operation. Clinical stages of the tumor: T1 for 42 cases, T2 for 12 cases. All cases were followed-up for 1 to 14 months, with a mean of 8.5 months. Tumor recurrences were found in 2 cases, no one had recurrence in situ. The tumor, submucosa and all muscular layers can be resected completely by 2 micron continuous wave laser transurethral partial cystectomy. Pathologic staging can be judged correctly. The umbilication raw surface were infiltrated by fibrous connective tissue and chronic inflammatory cells 1 week postoperation. The umbilication changed shallow and transitional epithelial cells began to cover it 1 month postoperation. The umbilication disappeared and transitional epithelial cells cover the raw surface 3 months postoperation. There was no difference between the raw surface and normal bladder mucosa.

CONCLUSIONS2 micron continuous wave laser for the treatment of bladder tumor can get the same clinical result as partial cystectomy. The pathologic staging can be judged correctly by the specimens.

Adult ; Aged ; Aged, 80 and over ; Cystectomy ; methods ; Female ; Humans ; Laser Therapy ; Male ; Middle Aged ; Neoplasm Staging ; Prospective Studies ; Treatment Outcome ; Urinary Bladder Neoplasms ; pathology ; surgery

OBJECTIVETo explore the feasibility and efficiency of extracting protein from bone tissue using trichloroacetic acid (TCA)-acetone precipitation method.

METHODSHydrochloric acid (HCL) decalcification method and TCA-acetone precipitation method were separately used for bone protein extraction. The efficiencies of these two methods were compared.

RESULTSTCA-acetone precipitation method had significantly higher extraction efficiency. Compared with HCL decalcification method, it had less pollution from bone marrow hematopoietic tissue. Protein band distribution was similar between these two methods.

CONCLUSIONTCA-acetone precipitation method is useful for bone proteomics research.

Acetone ; Animals ; Bone and Bones ; chemistry ; Chemical Precipitation ; Male ; Proteins ; isolation & purification ; Proteomics ; methods ; Rats ; Rats, Wistar ; Trichloroacetic Acid