ObjectiveTo investigate the role of caspase 3 in HMME-induced apoptosis in hypertrophic scar fibroblasts (HSFs).MethodsFibroblasts were obtained from 10 patients with untreated hypertrophic scar,and subjected to a primary culture.After 4 to 6 passages of culture,the HSFs were divided into 3 groups to remain untreated(control group),be treated with HMME followed by photodynamic therapy (HMME-PDT group),or the combination of HMME and Z-DEVD-FMK followed by photodynamic therapy (caspase 3 inhibitor group).At 12 hours after the therapy,HSFs were collected and immunofluorescence microscopy was used to observe the fluorescence intensity of caspase 3 after staining with fluorescein isocyanate (FITC) and popodium iodide (PI),flow cytometry was performed to determine the percentage of caspase 3-positive HSFs and apoptosis rate in HSFs after single staining with FITC and PI respectively.Results The fluorescence intensity of caspase 3 was weak in the control group and caspase 3 inhibitor group,but was strong in the HMME-PDT group.An increased percentage of caspase 3-positive HSFs was noted in the HMMEPDT group compared with the control group and caspase 3 inhibitor group(30.86％ ± 1.21％ vs.3.12％ ±0.28％ and 2.46％ ± 0.18％,t =19.92,21.76,both P ＜ 0.05).The apoptosis rate in HSFs was significantly higher in the HMME-PDT group and caspase 3 inhibitor group than in the control group(30.54％ ± 3.78％ and 10.46％ ± 2.15％ vs.2.45％ ± 0.22％,t =35.90,27.97,both P＜ 0.05),and higher in the HMME-PDT group than in the caspase 3 inhibitor group.ConclusionsThe apoptosis in HSFs induced by HMME-PDT is closely related to the activation of caspase 3,while caspase 3 seems to be dispensable for the apoptosis.

Objective To investigate the apoptotic effects of hypertrophic scar fibroblast (HSF) induced by HMME-PDT.Methods Fibroblasts were cultured from nontreated hypertrophic scars,and cells at passages 4-6 were used for the experiments (photosensitizer dose 4 μg/ml,λ630 nm,pow er density 10 mw/cm2,energy fluence 2.5 J/cm2).Morphological and biochemical changes in fibroblasts were assessed by Hoechst 33258 staining and fluorescence microscopy.The rate of apoptotic or necrotic cells was detected by flow cytometry (FCM) through double staining of Annexin V -FITC and popodium iodide (PI),respectively.Results Marked morphological features of cell apoptosis were viewed under the fluorescent microscope through Hoechst 33258 staining.The analysis of FCM indica ted that the apoptotic rate was significantly increased after HMME PDT [(34.82 ± I.42) ％ vs (3.12±0.28) ％,P＜0.05],and apoptotic rate was higher than necrosis rate [(14.65±1.02) ％ vs (34.82±1.42) ％,P＜0.05].Conclusions Low level exposure to 630 nm PDT mediated by HMME appears to induce fibroblast apoptosis.

Objectives To study health-related quality of life(HRQOL) of children with subclinical epileptiform electroencephalographic discharges.Methods Subjects were 30 children with subclinical epileptiform electroencephalographic discharges,50 children with clinical seizures,28 children without clinical seizures and 30 controls.Guardians completed a valid epilepsy-specific HRQOL questionnaire for children,the quality of life in children epilepsy questionnaire(QOLCE).Results Children with subclinical epileptiform electroencephalographic discharges have 8 subscales scores lower than control group,2 subscales scores higher than group with clinical seizures.Conclusions Quality of life of children with subclinical epileptiform electroencephalographic discharges is better than that of children with clinical seizures.However cognitive,behavior,daily life and social activity are impaired by subclinical epileptiform electroencephalographic discharges and lowered the HRQOL of children.

Objective To study the clinical outcomes and safety of immunoadsorption therapy for refractory autoimmune disease in children.Methods Three boys who suffered of severe autoimmune disease-one boy suffered of severe dermatomyositis and pulmonary infection; one suffered of severe anaphylactoid purpura with alimentary tract hemorrhage and entero ablation for intestinal perforation ; other one suffered of systemic juvenile idiopathic arthritis and severe prosopo-cellular tissue infection,macrophage active syndrome,were treated with blood immunoadsorption by resin immunoadsorbent of HA280.Then evaluated the clinical outcome of 3 cases,including symptom improvement,change of serum immune globulin,complement,enzyme of liver and heart,autoantibody.Results After the treatment of immunoadsorption,the symptom of 3 cases improved obviously; the sensitivity of the corticosteroids increased; autoantibody of antinuclear antibody (ANA) and anti-cyclic citrullinated peptide antibody (CCP) changed to negative; C-reactive protein (CRP) dropped (P ＜ 0.05) ; descending scale of IgM,IgA,C3,C4 increased (P ＜ 0.05) ; the normal scale of immunoglobulin didn't changed (P ＞ 0.05) ; besides aspartate aminotransferase (AST) dropped in the case of dermatomyositis,the other enzyme of liver and heart didn't changed.Conclusions The body could be restored quickly by the treatment of immunoadsorption together with the drug; CRP in the blood could be removed by immunoadsorbent of resin; 1 or 2 times blood immunoadsorption could not change the level of enzyme,but it need to do more on severe cases,especially those with poor organ function; for the safe of the treatment of immunoadsorption for the young age,low weigh and severe cases,the operative procedure should be critical care.

AIM:To assess changes of tear film function in patients after pterygium excision combined with amniotic membrane transplantation.
METHODS:Totally 126 patients with pterygium excision with amniotic membrane transplantation from January 2011 to November 2013 were entered in the study. The tear breakup time ( BUT) , the Schirmer I test ( SⅠt) and tear ferning test ( TFT ) were elevated in the patients before and after pterygium excision combined with amniotic membrane transplantation. The examnation times were 1d before surgey, 1wk, 1, 2mo after surgery. Operation eyes were studied group, while opposite healthy eyes as control group.
RESULTS: Compared with the control group, BUT and TFT were significantly different in the eyes with pterygium (P<0. 05); However, no obvious difference was detected in the results of SⅠt (P>0. 05). The results of BUT and TFT at 1mo after surgery in study group were significantly better than 1wk (P<0. 05), while no significant difference compared with 2mo (P>0. 05); The tear film stability in the study group at 1wk after surgery was still inferior to the control group (P<0. 05) and there was no significant difference at 1, 2mo after surgery (P all>0. 05). SⅠt results did not differ between the different examination times(P>0. 05).
CONCLUSION:Tear film stability was broken in the eyes with pterygium. Pterygium excision combined with amniotic membrane transplantation can obviously restore the tear film function into normal state, and the tear film function could reach steady-state 1mo after surgery.

·AlM:To evaluate the effect and safety of intrachamberal triamcinolone acetonide ( TA ) injection during cataract surgery on controlling postoperative inflammation and macular edema on diabetic patients.
· METHODS: Three hundred patients ( 300 eyes ) with type 2 diabetes who scheduled for cataract surgery were randomly divided into three groups: group A: 0.3%tobramycin/0. 1% dexamethasone eye drops and pranoprofen eye drops treatment for 1mo postoperatively;group B:intrachamberal injection of TA 1mg after cataract surgery, and 0.5% levofloxacin eye drops treatment for one month postoperatively; group C: intrachamberal injection of TA 2mg after cataract surgery, and 0.5%levofloxacin eye drops treatment for one month postoperatively.The main measurements included visual acuity, intraocular pressure ( lOP ) , corneal endothelial cell density, anterior chamber inflammation and the thickness of macula of the three groups.
· RESULTS:All cataract surgeries were done successfully by a single surgeon.The best corrected vision of group B and C was better than that of group A 1d, 1wk and 1mo postoperatively (P<0.05).The inflammation of anterior chamber of group B and C was milder than that of group A 1d and 1wk postoperatively (P<0.05).The average lOP of group C 1d postoperatively was higher than that of group A (P<0.05).There was no significant difference between group B and group C on the best corrected vision, anterior chamber inflammation and lOP at any time point.No significant change in the macular thickness was found in patients of group B and C before and after cataract surgery, while there was thicker macula in patients of group A 3wk and 1mo postoperatively when comparing with that of the baseline (P<0.05).There was no significant difference among three groups on corneal endothelial cell density at any time point.
· CONCLUSlON: lntrachamberal TA injection during phacoemulsification can effectively control postoperative inflammation, reduce the macular edema and accelerate the recovery of visual acuity.lntrachamberal TA 1mg has good effect and safety.

Objective To observe the clinical effect of intravitreal ranibizumab (IVR) combined with vitrectomy in treating proliferative diabetic retinopathy (PDR).Methods This is a prospective non-randomized controlled clinical study.A total of 62 patients (70 eyes) who underwent vitrectomy for PDR were enrolled and divided into IVR group (30 patients,34 eyes) and control group (32 patients,36 eyes).IVR group patients received an intravitreal injection of 0.05 ml ranibizumab solution (10 mg/ml) 3 or 5 days before surgery.The follow-up time was 3 to 18 months with an average of (4.5± 1.8) months.The surgical time,intraoperative bleeding,iatrogenic retinal breaks,use of silicone oil,the best corrected visual acuity (BCVA) and the incidence of postoperative complications were comparatively analyzed.Results The difference of mean surgical time (t=6.136) and the number of endodiathermy during vitrectomy (t=6.128) between IVR group and control group was statistically significant (P=0.000,0.036).The number of iatrogenic retinal break in IVR group is 8.8 ％ and control group is 27.8％,the difference was statistically significant (x2 =4.154,P=0.032).Use of silicone oil of IVR group is 14.7％ and control group is 38.9％,the difference was statistically significant (x2 =5.171,P=0.023).The incidence of postoperative vitreous hemorrhage in 3 month after surgery was 11.8％ and 30.6 ％ respectively in IVR group and control group.The differences were statistically significant (x2=3.932,P=0.047).The 6 month postoperative mean BCVA of IVR group and control group have all improved than their preoperative BCVA,the difference was statistically significant (t=4.414,8.234;P=0.000).But there was no difference between the mean postoperative BCVA of two groups (t=0.111,P=0.190).There was no topical and systemic adverse reactions associated with the drug after injection in IVR group.Conclusions Microincision vitreoretinal surgery assisted by IVR for PDR shorten surgical time,reduces the intraoperative bleeding and iatrogenic retinal breaks,reduces the use of silicon oil and the postoperative recurrent vitreous hemorrhage.But there was no significant relationship between vision improvement and IVR.

Objective To compare clinical outcomes in eyes with macula-off rhegmatogenous retinal detachments (RRD) with peripheral breaks managed by surgical protocols that result in either complete (CSFD) or partial subretinal fluid drainage (PSFD).Methods Following the clinical detection of a maculaoff RRD with peripheral retinal breaks,patients were offered the opportunity to enroll in the study,and those patients who signed the consent were evaluated for eligibility based upon the inclusion and exclusion criteria for this clinical study,and if fully eligible they were assigned prospectively to one of the two surgical designs (PSFD or CSFD,1∶ 1) using a random number table.Seventy-two eyes of 72 patients were enrolled and studied.Patients were treated with 25G plus vitrectomy,endolaser or transscleral cryopexy,either complete (n=36),or partial (n=36) subretinal fluid drainage,and 14％C3F8 (PFO) was used for intraocular tamponade.After surgery,all patients were kept in a supine position for 24 hours,and then in a clinically optimal position for 6-10 days.The study patients were examined at 1,3 and 6 months after surgery with thorough ophthalmic examinations.Macular optical coherence tomography (OCT) imaging was acquired in 1 month.Anatomical and visual outcomes as well as intra-operative and postoperative complications of the two groups were compared.Furthermore,the persistence of subfoveal fluid in OCT images and the symptoms of distortion at 3 months were measured and recorded.The primary study endpoint of anatomic retinal reattachment for each group was based upon the 6-month time-point.Results The preoperative baseline characteristics between the two groups were not significantly different.The single-operation success rates were 88.9％ and 91.6％ respectively for the CSFD and the PSFD groups (x2 =0.158,P＞0.05).The mean best corrected visual acuity (BCVA) at 6 month endpoint were 0.99± 0.52 minimum resoluation angle in logarithmic (logMAR) for the CSFD group and 1.07±0.34 logMAR for the PSFD group(t=0.580,P=0.564).The mean operative time was longer in the CSFD group (62.25± 4.32) minutes than that in the PSFD group (47.9 ± 5.0) minutes (t =0.580,P=0.564).seven of 29 (24.1％) phakic eyes in the CSFD group had lens injury during SRF drainage,and none of the 31-phakic eyes in the PSFD group sustained lens damage.Residual PFO was present in 6 of 36 CSFD cases (16.7％).Successful retinal reattachment after primary surgery was achieved in 33) PSFD eyes and in 32 CSFD eyes based upon OCT imaging at 1 month demonstrated reattached foveae with no residual subfoveal fluid.Among these patients,22 patients (62.5％) in the CSFD group and 23(69.7％) patients in the PSFD group reported distortion in the operated eye or/and a difference in image size between the two eyes at the 6 month visit (P=1.00).Conclusions Partial subretinal fluid drainage during pars plana vitrectomy for the repair of macula-off RRD with peripheral breaks is effective.The success rates are not statistically different.Additionally,PSFD procedures can simplify the surgery procedure,shorten operative time and,and to some extent,reduce the incidence of complications relevant to the CSFD approach.

Objective To observe the phosphorylation of Smad3 in hyperplastic scar fibroblasts (HSFs) induced by hematoporphyrin monomerthyl ether (HMME) followed by photodynamic therapy (PDT).Methods Fibroblasts were isolated from the hypertrophic scar tissues of 10 patients and subjected to culture in vitro.After 3-5 passages,the HSFs were divided into 4 groups:control group receiving no treatment,PDT group pretreated with HMME of 4 μg/ml followed by PDT,HMME group induced by HMME alone,and laser group irradiated with laser alone.Fluorescence microscopy was used to observe the expression of Smad3 after immunofluorescent staining with anti-Smad3 antibody,and Western blot to detect the expression of Smad3 and phosphorylated Smad3 in these HSFs.Paired t test was conducted to compare the difference in Smad3 and phosphorylated Smad3 expression between these groups.Results The total fluorescence intensity of Smad3 was similar between these groups,but the intranuclear fluorescence signal was significantly weaker in the PDT group than in the control group.The level of phosphorylated Smad3 was statistically decreased in the PDT group compared with the control group (0.20 ± 0.02 vs.0.92 ± 0.15,P ＜ 0.05),but no significant difference was observed between the HMME group and laser group (P ＞ 0.05).Conclusion PDT may inhibit the proliferation of HSFs via attenuating the phosphorylation of Smad3.

Aim To investigate the effects of diallyl di-sulfide( DADS) on G2/M arrest in Chk1/MGC803 and Chk2/MGC803 cells so as to establish stable human gastric cancer MGC803 cells with overexpression of Chk1/2 gene. Methods The colony formation, flow cytometry, RT-PCR and Western blot were used to de-tect the proliferation, cell cycle, and expression of Chk1/2 mRNA and protein, p-Chk1/2, CDC25C and cyclinB1, respectively. Results The colony formation showed that the colony forming efficiency in Chk1/MGC803 and Chk2/MGC803 cells treated by 30 mg· L-1 DADS was lower than in control group and vector group ( P <0. 05 ) . Flow cytometry demonstrated that 41. 3%, 57. 4%, 68. 9% and 42. 9% of G2/M cells in Chk1/MGC803 were increased than in MGC803 and Chk2/MGC803 , respectively after treated by DADS in 12,24, 36 and 48 h(P <0. 05). At the same time, RT-PCR disclosed that expression of Chk1 and Chk2 mRNA had no marked change. Western blot showed that total proteins of Chk1 and Chk2 and p-Chk2 had invisible change, but expression of p-Chk1 was up-reg-ulated, and CDC25C and cyclinB1 were down-regula-ted time-dependently in Chk1/MGC803 cells ( P <0. 05 ) . Conclusion DADS arrests MGC803 cells at G2/M by increasing p-Chk1 expression to cause down-regulation of CDC25C and cyclinB1 simultaneously.