Background Protein kinase B (Akt) is the center of multiple cellular signaling pathways,and it participates in the regulation of cell function,such as proliferation,migration,and metabolism of cells.Phosphoinositide 3-kinase (PI3K) promotes activation of Akt and therefore triggers many signal pathways.PI3K inhibitor can silent Akt,but whether it can affect the biological behavior of lens epithelial cells (LECs) during the posterior capsular opacity (PCO) is worthy of investigation.Objective This study was to explore the effect of LY294002,a PI3K inhibitor,on Akt activation in human LECs.Methods Human LECs strain,HLEC-B3 cells,were cultured and passaged.The cells were incubated to 96-well plate for 24 hours,then LY294002 was added with the final concentration at 0,10,20,30,40,50,60,70,80 μmol/L,respectively.After incubation for 24 hours,cell counting kit-8 (CCK-8) was used to detect the inhibitory rate of cell proliferation.Transforming growth factor-β2 (TGF-β2) of 10 μg/L was added in the medium of cells as TGF-β2 group,TGF-β2 + LY294002 (20 μmol/L) was used to coculture the other group of cells,and the cells without TGF-β2 and LY294002 served as the control group.Phosphorylation of Akt (p-Akt) in the cells was detected by laser scan confocal immunofluorescence microscope.The expression level of p-Akt was evaluated using Western blot.Results CCK-8 assay showed that the A value of the cells was gradually reduced with the increase of LY294002 concentration (Fgroup =9.72,P =0.00),but the A value was significant raised along with the lapse of time (Ftime =1737.54,P=0.00).Confocal immunofluorescence revealed that a little of p-Akt was expressed in the control cells.After induced by TGF-β2,lots of red fluorescence of p-Akt was seen in cells around the cell membranes.But in the TGF-β2+LY294002 co-culture cells,the fluorescence of Akt was much weaker.Western blot showed that the expression level of p-Akt was 0.91±0.08,1.48±0.13 and 0.95±0.19 in the control group,TGF-β2 group and TGF-β2 +LY294002 group,respectively,with a significant difference among the three groups (F =15.04,P =0.00).Conclusions LY294002 can inhibit the activation of Akt induced by TGF-β2.LY294002 may have utility in the prevention and treatment of PCO.

Objective To investigate the clinical significance of CD55,CD59 and Aeromonas hydrophila toxin variant (FLAER) in the diagnosis of anemia and paroxysmal nocturnal hemoglobinuria (PNH).Methods Collected 30 healthy controls,22 cases of PNH,33 cases of aplastic anemia (AA),37 cases of iron deficiency anemia (IDA),45 cases of megaloblastic anemia (MA),30 cases of hemolytic anemia (HA) and 31 cases of myelodysplastic syndrome (MDS) from January 2009 to March 2017,CD55,CD59 and FLAER negative cell ratio of peripheral blood neutrophil of them were detected by multipa rameter flow cytometry.Results The detection rates of FLAER in PNH,AA and MDS groups were higher than those of CD55 and CD59,but there was significant difference in AA (x2 =7.759,5.518,P=0.005,0.019＜0.05).The average CD55,CD59 and FLAER deletion rate in PNH and AA group were significantly higher than those in normal control group and other groups (t=2.163～17.890,P=0.000～0.038＜0.05).The number of FLAER in PNH group was higher than CD59 and CD59 was higher than CD55 with the statistically significant difference (t=2.503 ～ 6.308,P=0.000 ～0.016＜ 0.05).Conclusion CD55,CD59 and FLAER have important value in the diagnosis of PNH and differential diagnosis with other anemia diseases,and can also be used to detect the presence of MDS and AA in patients with PNH.FLAER outperforms CD59,CD59 outperforms CD55.

Background The pathogenesis of age-related cataract is associated with the apoptosis of lens epithelial cells (LECs) caused by oxidative stress.Previous studies showed that intracellular focal adhesion kinase (FAK) pathway can be activated by H2O2 in vitro,which induced apoptosis of cells.To investigate the effect of oxidative on FAK expression in LECs is one of important studies in the prevention of age-related cataract.Objective This study was to investigate the expression and function of FAK in human LECs treated by H2O2.Methods Human LECs strain (HLECs-B3) were cultured in vitro in the low glucose DMEM with 10％ fetal bovine serum.Different concentrations (0,30,50,70,100,300,500,700,1000 μmol/L) of H2O2 were added into the culture medium for 24 hours.The survival rate of the cells was detected by Cell Counting Kit-8 (CCK-8) assay.Cell morphology as well as the expression and distribution of FAK in the cells were observed by immunofluorescent staining under the laser confocal microscope.Apoptosis was observed by hoechst33258 staining,and Western blot assay was used to quantitatively detect the expression and phosphorylation of FAK.Results The survival rate of the cells was (1.00±0.03) ％,(1.24±0.03)％,(1.36±0.24) ％,(0.93±0.02)％,(1.75±0.19)％,(1.37±0.18) ％,(0.64±0.01)％,(0.59±0.11)％,(0.14±0.05)％ in 0,30,50,70,100,300,500,700,1000 μmol/L H2O2 groups,with a significant difference among them (F =95.30,P =0.00).The survival rates of the cells in the below 300 μmol/L H2O2 groups were significantly higher than those in the 0 μmol/L H2O2 group,and survival rates of the cells in the above 500 μmol/L H2O2 groups were significantly lower than those in the 0 μmol/L H2O2 group(all at P＜0.05).After H2 O2 treatment for 24 hours,HLECs-B3 cells transformed from polygon shape to spindle shape and extended pseudopodiums,meanwhile the green fluorescence for FAK exhibited in the cytoplasm.Cell apoptosis was found in the 1000 μ mol/L H2O2 group.Western blot assay revealed that the expressing levels (grey scale) were significantly different among the various groups (F=28.08,P=0.00),and FAK expressing levels in the below 300 μmol/L H2O2 groups were significantly higher than those of the 0 μmol/L H2O2 group; while the expressing levels in the above 500 μmol/L H2O2 groups were lower than those of the control 0 μmol/L H202 group (all at P＜0.05).After treated by different concentrations of H2O2,the phosphorylation level of intracellular FAK (p-FAK) was significantly higher in 3 hours group than that in 30 minutes group (all at P＜0.05).Conclusions H2 O2 can affect the survival,proliferation and morphology of human LECs by activating the intracellular FAK pathway,indicating that FAK may play roles in the regulation process of cell biological behavior.

Background Surgical induced astigmatism induced by a 5.5mm of superior scleral tunnel incision is a key cause of influencing vision outcome in the high myopic eyes after implantation of Artisan phakic intraocular lens(IOL).Objective This study was to evaluate the surgical induced astigmatism through a 5.5mm superior scleral tunnel incision during the implantation of Artisan phakic IOL.Methods This was a case observational study.The clinical data of 202 eyes of 111 patients with Artisan IOL implantation for high myopia from October 2004 through October 2008 was collected.The patients were followed-up for 12 months.The uncorrected visual acuity(UCVA),best spectacle-corrected visual acuity(BCVA),spherical equivalent(SE),cylinder equivalent(CE) and cylinder axis were examined before surgery and postoperative 1,3,6,12 months.The patients were divided into with rule group and against rule group according to the cylinder axis.Surgery-induced astigmatism value was calculated using Holladay vector analysis formula.This clinical trial was approved by the Ethics Committee of Guangdong General Hospital.Written informed consent was obtained from each patient prior to the surgery.Results The UCVA of 195 eyes(94.1%) was 0.5 or better in postoperative 12 months,and that of 172 eyes(85.1%) showed the improvement of BCVA during the follow-up duration.The mean CE values were reduced in 1,3,6,12 months after surgery in comparison with before surgery(t=-4.30,P=0.00;t=-2.27,P=0.01;t=-2.04,P=0.04;t=-2.79,P=0.01).Vector analysis of total surgically induced astigmatism revealed that the mean incision-induced astigmatism value was +1.94D,+2.26D,+2.29D,+2.25D at 171°,170°,181°,175° in 1,3,6,12 months after surgery.The surgery-induced astigmatism was(+1.97±1.84)D,(+2.25±1.75)D,(+2.27±1.76)D,(+2.24±1.75)D in with rule group at postoperative 1,3,6,12 months respectively;while those in against rule group was(+1.75±1.88)D,(+2.35±1.74)D,(+2.38±1.76)D,(+2.34±1.74)D respectively.No significant differences were found between the two groups(t=0.54,-0.29,-0.27,-0.29;P=0.59,0.78,0.79,0.78).Conclusion Implantation of Artisan IOL is an effective approach for correction of high myopia.A 5.5mm superior scleral tunnel incision in operation induces a +2.25D astigmatism at 175° meridian.

OBJECTIVETo explore the clinical effects of Dynesys system for the treatment of multiple segment lumbar degenerative disease.

METHODSA total of 28 patients with lumbar degenerative disc disease treated with Dynesys system from December 2008 to May 2011 were retrospectively reviewed. There were 16 males and 12 females, aged from 27 to 75 years old with an average of 49.1 years. Thirteen patients with multiple segmental lumbar intervertebral disc protrusion, including L3-L5 in 7 cases, L2-L4 in 1 case and L4-S1 in 5 cases. Fifteen patients with multiple segmental lumbar spinal stenosis, including L3-L5 in 10 cases, L4-L5 in 4 cases and L2-S1 in 1 case. The symptoms of lumbago and (or) intermittent claudication in all patients were treated with conservative treatments for more than 6 months and these methods did not work. Visual analogue scale (VAS) was used to analyze the lumbar and leg pain, imaging data were used to measure the intervertebral space height and intervertebral motion of fixed segment and upper adjacent segment, Oswestry Disability Index (ODI) was used to evaluate the clinical effect.

RESULTSAll operations were successful and the patients were followed up from 38 to 65 months with an average 50.6 months. At final follow-up, ODI and VAS of the low back pain and leg pain were (25.10±6.52)%, (1.25±0.70) points and (1.29±0.89) points, respectively and were decreased compared with preoperative (P<0.05). Postoperative intervertebral space heights were increased and intervertebral motions were decreased in fixed segment compared with preoperative (P<0.05). There were no significant differences in intervertebral space heights and intervertebral motions of upper adjacent segment between preoperative and postoperative (P>0.05).

CONCLUSIONDynesys system may obtain long-term clinical curative effect in treating multiple lumbar degenerative disease. It can partially preserve the intervertebral motions of the fixed segments, have little effect on adjacent segments. The long-term clinical effect of Dynesys still need longer time follow-up observation.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Degeneration ; pathology ; surgery ; Joint Instability ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Visual Analog Scale

Objective To investigate the effects of pulsed electromagnetic fields(PEMFs)on proliferation and differentiation of endothelial progenitor cells(EPCs).Methods EPCs were isolated from rat bone marrow by density gradient centrifugation.EPCs were exposed to PEMFs from the 5th day to the end of culture.MTT was used to measure the proliferation of EPCs.The expression ofⅧ-related antigen and NOS_3 was evaluated by flow cytometry. Results Compared with the control,the proliferating ability of EPCs exposed to PEMFs was stronger;the number ofⅧ-related antigen and NOS_3 positive cells increased significantly in EPCs exposed in PEMFs.Conclusion PEMFs promotes the proliferation and differentiation of rat bone marrow EPCs.

The thermotolerant bacteria which was isolated from the Apple Juice Concentrate(AJC) was investigated by comparison with the standard strain of Aliyclobacillus acidoterrestris DSM3922 using Kirin-medium acidified with malic acid.The results shows the two polluted bacteria isolated from AJC can grow under the temperature of 21?C～55?C and pH of 2.4～6.2,which was corresponded with the characteristics of the thermo-acidiphilic Alicyclobacillus spp..Further more,the cell morphology,colony morphology,cultural characteristics and physiological characteristics tests indicated the two isolated strains of thermotolerant bacteria have obviously similar characteristics with the standard strain of Aliyclobacillus acidoterrestris DSM3922.

Objective To study the mutagenic action of antiepileptic drugs(AEDs), and find an effective way to prevent the mutagenesis induced by AEDs,by observing the effects of AEDs on serum folic acid(FA) level and sister chromatid exchange (SCE) frequency in epileptic children.Methods Ninty epileptic children were divided into different groups on the basis of the different drugs they had taken, then detected the two indexes at different time points.Results The serum FA level and SCE frequency of the patients significantly decreased and increased after they took carbamazepine (CBZ) and valproic acid (VPA)respectively. The two indexes went back respectively when supplied with FA.Conclusions Both CBZ and VPA possess mutagenic action, yet nitrazepam does not.FA may help repair the chromosome damage and reduce the mutagenesis effects.

Objective To study the clinical features,diagnosis and treatment of eosinophilic granuloma of bone(EGB).Methods Twelve cases(8 male and 4 female)of EGB and their medical record,imaging examinations,follow-up data were reviewed,and the lesion sites,symptom and clinical features were analyzed with reference to relative literature.Results Nine unifocal cases were found in the cervical spine(3 cases),the thoracic spine(1 case),the lumber spine(1 case),the femur(1 case),the metacarpal bones(1 case),and the ilium(2 cases).Three multifocal cases were found in the thoracic and lumber spine(1 case),in the femur,the multiple ribs,the skull,the scapula(1 case)and in the skull,the femur,the ilium(1 case).The most common presenting symptom was pain at the lesion site and restricted motion was obviously in the cervical lesion.The case of the thoracic lesions was found neurologic deficit.Osteolytic destruction was found in the radiologic examination in EGB and the periosteal reaction was found in the long bone lesions,and vertebra plana was considered as typical cha-racteristics in the spine lesion.The lesions not in the spine were diagnosed by biopsy,exclusive methods and close following-up were performed in the spine lesions.Two unifocal were healed spontaneously and three were performed with curettage with one bone grafting.Multifocal and the lesions in the spine underwent chemotherapy.All cases were followed up and no recurrence was found.Conclusions EGB is commonly solitary and not seldom in the spine.Topical pain may be chief complaint.Osteolytic image was found in the radiologic examination.Biopsy and exclusive methods combining closely follow-up were used for diagnosis.EGB is self-limited observation,so curettage or chemotherapy can be used in treatment,and prognosis is good.

Background Some aboard studies showed that polyatomic pathway play an important role in the microsvasculopathy of type 2 diabetes,and sorbitol dehydrogenase (SDH) gene is involved in the pathogenesis of diabetic retinopathy(DR).There is no relevant research at home up to now.Objective This study was to investigate the correlation of sorbitol dehydrogenase (SDH) G-888C gene polymorphism with diabetic retinopathy (DR) in type 2 diabetes mellitus.Methods The SDH genotypes were determined by polymerase chain reactionreaction fragment length polymorphism (PCR-RFLP) in 187 patients with diabetes and 123 normal contrels.Patients were divided into two groups depending on the presence or absence of DR(DR:n=118;NDR:n=69).The frequencies of SDH genotype and allele were assayed and compared among these groups.This study was approved by Ethic Committee of Guangdong General Hospital.Written informed consent was obtained from each individual prior to any relevant medical procedure.Results The disease course in the DR group was significantly longer than that in NDR group(t=2.070,P=0.042),and other clinical features in both groups were non-significant (all P>0.05).The genotype frequencies in the DR group,NDR group and normal control group were 24.6%,8.7% and 8.1%,respectively,and frequencies of the G allele were 42.4%,25.4% and 29.7%,showing statistically significant differences among these three groups.The GG genotype and G allele frequencies were significantly higher in the DR group than those in the NDR group and normal control group (GG:P=0.007,P=0.001;G:P=0.001,P=0.004).There were no significant differences in the frequencies of the GG genotype and G allele between the NDB group and normal control group ( P>0. 05) as well as the proliferative DR group and non-proliferative DR group (P>0.05).Conclusion SDH G-888C gene polymorphism is associated with the development of diabetic retinopathy in southern Chinese.