Objective To analyze the protein expression changes of retina in diabetic mice using isobaric tags for relative and absolute quantitation (iTRAQ) approach and to study proteins of apoptosis.Methods 8 diabetic mice were chosen as the diabetic model group (DM group),8diabetic mice as the normal control group.The animals were housed in wire-bottomed cages and received normal pellet chow and tap water in a constant environment.After 10 weeks,all mice were killed,and their retina were dissected.After hematoxylin and eosin(H&E) staining,the sections of retina were examined using light microscopy.The changes of protein expression in retina were studied using iTRAQ approach.Expression of apoptosis associated proteins was analyzed using ingenuity pathway analysis (IPA).Results Compared with control group,mice retina in DM group developed looser structures,tissue edema and obvious telangiectasia under light microscopy.Using iTRAQ approach,a total of 348 differential proteins were identified.Among those proteins,16 proteins were related with apoptosis,including Ataxin-10,Protein NDRG1,mucin-4,Aquaporin-1 and annexin A4,etc.There were 8 apoptosis-related proteins in retina with up-regulation,and the other 8 proteins with down-regulation in the DM group.The relationship between these proteins were analyzed and charted by IPA.Conclusions Apoptosis may be involved in the development of diabetic retinopathy.The identification of the apoptosis-related proteins will be helpful for the further study.

AIM: To compare and analyze the stereopsis of intermittent exotropia children under the different backgrounds of dynamic stimuli and static stimuli.
METHODS: We collected 56 children ( male 26, female 30 with intermittent exotropia at the age from 5y to 12y and examined their stereopsis under the different backgrounds of dynamic stimuli and static stimuli using a multidimensional sense perception training software. The differences between the dynamic stereopsis and static stereopsis were compared.
RESULTS: Totally 17 cases ( 30%) had both dynamicstereopsis and static stereopsis, 39 cases ( 70%) had either dynamic or static stereopsis deficit, only 10 cases ( 26%) had dynamic stereopsis, 25 cases ( 64%) static stereopsis left and 4 cases ( 10%) were without any form of stereopsis. The positive rate of dynamic stereopsis was better than the positive rate of static stereopsis, with statistical significance (P<0. 05).
CONCLUSION: Dynamicstereopsis is better than the static stereopsis to intermittent exotropia children.

Objective To study the effects of matrine on accumulation of ? globin mRNA and induction of erythroid differentiation in K562 cells in vitro.Methods K562 cells were cultured for 6 days with different concentration of matrine,viable cell counts were determined by trypan-blue dye exdusion test. Erythroid differentiation was evaluated by percentage of benzidine-positive cells at different days after culture. Morphological changes were observed under microscope after Wright-Gimesa staining; ? globin mRNA was quantitative by real time quantitative reverse transcript polymerase chain reaction(RT-PCR).Results Different concentrations of matrine inhibited proliferation of K562 cells in dose-dependent manner; otherwise, K562 cells were successfully induced by erythroid differentiation with matrine. After treatment with matrine, percentage of benzidine-positive cells significantly increased from 0.7% to 15.7% and characteristic changes of erythroid differentiation in the cell morphology were observed, G? globin mRNA had a preferential increase (2.7 fold)in K562 cells. Conclusions Matrine accumulation G? globin mRNA and induced erythroid differentiation of K562 cells. The results provides an experimental evidence for the pharmacological therapy of hematological diseases associated with a failure in the expression of normal ? globin genes.

Aim To ascertain whether defibrase has the protective effect against reperfusion injury after cerebral ischemia.Methods 70 renovascular hypertensive rats(RHR) were randomly divided into defibrase group, control group and sham-operated group.Reversible middle cerebral artery occlusion(MCAO) models were produced by the modified. Longa's method,and reperfusion was begun 2 hours after occlusion.Rats in the defibrase group were given defibrase 10 U?kg-1 body weight via femonal intraveneous injection, and in the control group with the same amount of saline. The brain pieces were processed by TTC and HE staining and the infarct size,brain microvessels damage and secondary bleeding were compared between the two groups. Results The volume of infarction in the defibrase group was obviously smaller than in the control group, the damage of brain microvessels was less severe, and the bleeding lesions under optical microscope were less than in the control group. Conclusion Defibrase has protective effect against reperfusion injury post cerebral ischemia.

Objective To retrospectively analyze and compare the curative outcome of hematopoietic stem cell transplantation (HSCT) from HLA identical siblings vs intensive immunosuppression therapy (IST) for severe aplastic anemia (SAA).Methods From January 2008 to December 2010,41 patients with severe aplastic anemia were treated with related HSCT (n =14) or IST (n =27) which combined antithymocyte globulin (ATG) with cyclosporine-A (CsA) therapy.Results All the patients receiving HSCT reached complete response.Among the patients receiving IST,21 patients could be responsive to the therapy,and 2 patients died.There was significant difference in the response rate between HSCT group and IST group (100 ％ vs 77.8 ％,P＜0.01 ).Conclusion With the improvement of HSCT technology,the curative outcome of HSCT from HLA identical siblings for SAA is much better than IST.

Objective To analyze the outcome of idarubicin-intensified myeloablastive conditioning regimen in allogeneic peripheral blood stem cell transplantation (allo-PBSCT) in patients with myelodysplastic syndromes (MDS). Methods From August 2004 to July 2009, 12 patients with MDS were treated with alIo-PBSCT following the idarubicin-intensified conditioning regimen. The conditioning regimen was idarubicin (15 mg/m~2), continuous intravenous infusion for 20 h, days-12 to-10; busulfan (0.8 mg/kg), intravenous infusion every 6 h, days-6 to-4; cyclophosphamide (1.8 g/m~2), intravenous infusion every 6 h, days-3 to-2; cyclosporine A combined with short-term methotrexate was used for the prophylaxes of acute graft versus host disease (aGVHD). Results All twelve patients achieved Trilineage engraftment, and were well tolerated to this regimen. Eight patients survived, and the overall survival was 66.7%, disease-free survival (DFS) 58.3%. Two patients relapsed. OS for neither WHO subgroups nor IPSS subgroups had statistically significant difference. Conclusion Allo-PBSCT with idarubicin-inteusified conditioning regimen is an effective treatment with reduction of the relapse rate for MDS patients.

AIM: To explore the structural changes of brain microvasculature and mechanism in microvascular lesion after focal cerebral ischemia with reperfusion. METHODS: Using the techniques of immunohistochemical staining, in situ hybridization,optical microscopy and transmission electron microscopy, the expression of uPA, uPA mRNA, and changes in miocrovascular structure were examined in ischemic focus and perifocal areas after focal cerebral ischemia 2 hours with various time points of reperfusion in stroke-prone renovascular hypertensive rats (RHRSP). RESULTS: The brain edema and hemorrhage were severe 12 hours to 3 days after reperfusion. Ultrastructural change showed that the damage characterizations of the basement membrane were degradation, defection, and exfoliated of basement membrane, while uPA, which attack the basememt membrane around cerebral capillaries and extra-cellular matrix, and uPA mRNA expression increased significantly in ischemic and perifocal areas 12 hour to 3 day after reperfusion. CONCLUSION: The main pathologic mechanism of brain edema and hemorrhage after cerebral ischemia with reperfusion may result from the basement membrane lesion of brain microvasculature. The increase in the expression of uPA in reperfusion area may be the main cause of the basement membrane lesion .

The roles of voltage-dependent K(+) channels during activation and damage in alveolar macrophages (AMs) exposed to different silica particles were examined. Rat AMs were collected by means of bronchoalveolar lavage, and were adjusted to 5x10(5)/mL. After AMs were exposed to different concentrations (0, 25, 50, 100, 200 mug/mL) of quartz particles and 100 mug/mL amorphous silica particles for 24 h, the voltage-depended K(+) current in AMs was measured by using patch clamp technique. Meanwhile the leakage of lactate dehydrogenase (LDH) and the viability of AMs were detected respectively. Patch clamp studies demonstrated that AMs possessed outward delayed and inward rectifying K(+) current. Exposure to quartz particles increased the outward delayed K(+) current but it had no effect on inward rectifier K(+) current in AMs. Neither of the two K(+) channels in AMs was affected by amorphous silica particles. Cytotoxicity test showed that both silica particles could damage AM membrane and result in significant leakage of LDH (P<0.05). MTT studies, however, showed that only quartz particles reduced viability of AMs (P<0.05). It is concluded that quartz particles can activate the outward delayed K(+) channel in AMs, which may act as an activating signal in AMs to initiate an inflammatory response during damage and necrosis in AMs induced by exposure to quartz particle. K(+) channels do not contribute to the membrane damage of AMs.

Background Rapamycin(RAPA)is a specific inhibitor of the mammalian target of rapamycin (mTOR).Researches showed that RAPA inhibits the proliferation of lens epithelium cells(LECs)and tumor cells and induces apoptosis of tumor cells.To investigate whether rapamycin has the inhibitory effect on retinal pigment epithelium(RPE)cells is very important for the prevention and management of proliferative vitreoretinopathy (PVR).Objective This study was to investigate the effects of RAPA on the proliferation and apoptosis of human RPE cells in vitro.Methods Human RPE cells(D407 strain)were cultured and passaged and then divided into regular culture group(blank control group),DMSO control group(0.1‰ DMSO +regular culture),and different concentrations RAPA-treatment groups(5,10,20,40,80,160,320 nmol/L).The proliferation(A490)of human RPE cells was detected using MTT,and the inhibitory rates of RAPA on the proliferation of RPE cells were calculated and compared among different groups at 12,24 and 48 hours.The apoptosis rates of the cells were analyzed among various groups by Hoechst staining after 12,24,48 hours.Results The inhibitory rates of RAPA on RPE cells were significantly different among various groups(F=484.451,P＜0.01)and evidently elevated in 20-320 nmol/L RAPA groups compared with DMSO control group(P ＜ 0.01).The inhibition of RAPA on the cells was considerably enhanced as the lapse of time(F=232.262,P＜0.01)with more dominant effects in 24 and 48 hours compared to 12 hours after addition of RAPA(P＜0.05-0.01).Compared with blank control group and DMSO control group,the apoptotic rates of the cells were evidently increased in 12,24,48 hours in 10 nmol/L RAPA group(all P＜0.05),and higher cellular apoptotic rates were found in 20-320 nmol/L RAPA groups(all P＜0.01).The alteration of cellular apoptotic rate showed a gradually incremental trend as the acting time of RAPA(F =625.584,P＜0.01).Karyorrhexis and mass-like density staining and chromatin substance were seen in RPE cells under the fluorescence microscope in ≥ 10 nmoL/L RAPA groups.Conclusions RAPA suppresses the proliferation and induces the apoptosis of human RPE cells in concentration-and time-dependent manner in vitro.

Objective With the method of detecting hepatitis B virus deoxyribonucleic acid(HBV-DNA) gene in breast milk,in ordered to diagnose whether there is HBV in breast milk in those mothers whose HBsAg are positive.Methods Collected breast milk from 187 mothers whose HBsAg were positive,and complified HBV-N gene in breast milk by Nested-Polymerase chain reaction (Nested-PCR).Results Nested-PCR could detect HBV-DNA in breast milk,and the positive rate was 3.2%.Conclusion The method of detecting HBV-N gene in breast milk by Nested-PCR can detect HBV-DNA in breast milk,and it can be an laboratory evidence for whether breast feeding or not.