BackgroundRetinal pigment epithelial (RPE)cell senescence damage the metabolism of photoreceptor,leading to retinal dysfunction and loss of vision.To understand RPE cell senescence mechanism will contribute to the study of age-related macular degeneration ( AMD).ObjectiveThe present study was to prepare the ageing RPE cell model with passage and explore its potential mechanisms.MethodsThis study was approved by the Ethic Committee of Qingdao University Medical College,and the informed consent was obtained from each gravida.Six human eyeballs were obtained from artificial labor fetusl with the gestational age 16-28 weeks.RPE cells were isolated,cultured and passaged in vitro to establish the cell replicative aging model.The third to twelfth cells were collected to be used to this experiment.Human keratin was used to identify the cells by immunochemistry,and MTT method was utilized to assess the proliferation and viability of different generations of cells as the A490 value.The cellular cycles and transmembrane potential (△ψm)of mitochondrion (△ψm) with passage were detected and compared using Flow Cytometry. Results Cultured and passaged cells showed the hexagon in shape with the melanin in 1-2 generations of cells and presented with the brown staining in cytoplasm for human keratin.The melanin was absent in the third generation cells.Vibrant growth statues were seen from the 3-6 generations cells and thereafter the proliferation ability reduced.The cells of G0/G1 phase were gradually elevated with the passage from 3 - 12 generations with a percentage of 68.40％ in the third generation of cells to 87.33％ in the twelfth generation of cells,showing a significant difference among various generations ( F =180.43,P =0.00),and that of the sixth,ninth and twelfth generation of cells was significant higher than the third,sixth and ninth generation respectively (t =4.002,P＜0.05 ; t=12.885,P＜0.01 ;t=16.387,P＜0.01 ).MTT assay showed that of RPE cells were significantly declined with the passage ( F =38.77,P =0.00),and the A490 value of the ninth,twelfth generations of cells was considerably lower in comparison with sixth and ninth generation respectively ( t =5.991,11.983,P＜0.01 ).From 3 through 12 generations of cells,the staining intensity of rhodamine 123 was gradually decreased ( F =121.68,P =0.00 ),and the staining intensity in the sixth,ninth and twelfth generation of cells was significant lower than that the third,sixth and ninth generation respectively(t=6.918,7.620,11.207,P＜0.01 ).Conclusions A replicative aging model can be successfully created by the passage in vitro using human fetal RPE cells.The reduce of transmembrane potential and damage of mitochondria might be one of mechanisms of senescence of RPE cells.

Objective To investigate the VEGF-C on lymph node metastasis of b-reast cancer.Methods A group of immune PV9000 law on human breast infiltrating ductal breast cancer and fibroids,breast disease organizations paraffin sections were detected on the expression of VEGF-C status.Results 20 patients with breast fibroids and breast disease were negative VEGF-C,80 cases of breast cancer VEGF-C positive rate was 72.5%(58/80);VEGF-C exists primarily on the cancer cells and normal breast fibroids and in the cytoplasm of breast disease is not the expression of VEGF-C.Conclusion Breast cancer cells in the VEGF-C,the expression of close ties with lymph node metastasis.Detection of VEGF-C expression in human breast cancer may predict lymph node metastasis,and therapeutic value.

Objective To evaluate if the enteral nutrition have effect on the immune function and in- flammatory reaction after operating about gastric carcinoma.Methods 58 postoperative patients suffering from stomach cancer and colon cancer were randomly divided into the EN group and the TPN group.On the first postoperative day,nutrition fibre were given via nasal intestinal tube,increasing the capacity and drop- ping speed day by day until patients can eat liquid diet.While patients in the TPN group didn't eat anything until enterokinesia completely recovered.Observing on preoperative day 1 and on postoperative day 3 and day 8 respectively to check IgA,IgG,IgM,C3,C4,CRP,LYM,LYM%,TP,ALB,PA.Results The results showed that on the postoperative day 8,the target ascension extent was higher than that in the PN group.The statistical significance was very obvious(P

AIM: To study the application of macular ganglion cell complex (mGCC) and peripapillary retinal nerve fiber layer thickness (pRNFL) measured by optical coherence tomography (OCT) in the early diagnosis of glaucoma.METHODS: Case-control study.Eighty-six subjects, including 30 eyes in normal subjects, 27 eyes in suspected primary open angle glaucoma, 29 eyes in primary open angle glaucoma were enrolled in this study.The thickness of mGCC and pRNFL were measured by OCT.The area under the receiver operating characteristic(AROC) curve at fixed specificities were calculated for each parameter.RESULTS: There were significant differences in mean pRNFL thickness, superior pRNFL thickness and inferior pRNFL thickness between normal group, suspected glaucoma group and early glaucoma group (P=0.001, 0.004, 0.011).The mean mGCC thickness, the thickness of the top mGCC, the thickness of the lower mGCC were statistically significant (P=0.008, 0.002, 0.003);the difference of general loss of volume (GLV) and focal loss of volume (FLV) between the three groups was statistically significant (P=0.002).Compared with the normal group, all the pRNFL and the mGCC parameters were higher in the suspected glaucoma group, and the FLV had the highest AROC (0.801), all the remaining AROC was >0.700 except above Prnlf (0.688).Compared with the normal group and the early glaucoma group, all the pRNFL and the mGCC had higher AROC, average mGCC was hightest(0.804), all parameters AROC were >0.700 except mean pRNFL (0.683).In suspected glaucoma group, 58% patients had abnormal mGCC thickness and 23% had abnormal pRNFL thickness;in early glaucoma group, 98%patients had abnormal mGCC thickness and 90% had abnormal pRNFL thickness;in normal group, 93%patients had abnormal mGCC thickness and 93%had abnormal pRNFL thickness, the correlation between the three groups was statistically significant (x2=12.11, P<0.05).CONCLUSION: OCT measurement of mGCC thickness and pRNFL thickness in early glaucoma have good diagnostic ability;mGCC thickness measurement can be used as an effective method for early diagnosis of glaucoma.

Objective:To evaluate the efficacy of laparoscopic intrafascial hysterectomy.Methods:Thirty women underwent laparoscopic intrafascial hysterectomy(GroupⅠ),29 women underwent open intrafascial hysterectomy(GroupⅡ) and 30 underwent abdominal hysterectomy(Group Ⅲ) were compared.The operating time,blood loss,postoperative pain,postoperative morbidity,time to first flatus and emiction were analyzed.Results:The blood loss was in Group I(61.00?21.95)ml,respectively,it was significantly lower than those in the GroupⅡ(121.03?53.61)ml and Group Ⅲ(101.00?21.07) ml(P0.05).Conclusion:Laparoscopic intrafascial hysterectomy is truly a minimally invasive surgery,and was associated with significantly less intraoperative bleeding,shorter hospital stay with less morbidity and shorter convalescence period.This remains the obvious advantages of laparoscopic in comparison with abdominal hysterectomy.

Objective To investigate the clinical application of ultrasound biomicroscopy in the treatment of congenital corneal opacities.Methods Medical records of 20 eyes (15 patients) with congenital corneal opacity treated at our hospital from July 2004 to November 2011 were retrospectively reviewed.Best corrected visual acuity testing,intraocular pressure testing,slit-lamp anterior segment examination,fundus examination,slit-lamp microscopic photography,B scan examination,and ultrasound biomicroscopy were performed for analysis of complications of congenital corneal opacity and selection of surgical approaches.Results The ultrasound biomicroscopic examination showed that 5 eyes had no Descemet's membrane and corneal endothelium,20 eyes had anterior synechia,5 eyes had aniridia,3 eyes had loss of lens cortex,13 eyes had cataract,14 eyes had closed angle,and 3 eyes had pupillary membrane.14 of 20 eyes received surgical treatment,including penetrating keratoplasty combined with cataract extraction and trabeculectomy (5 eyes),penetrating keratoplasty combined with pupil angioplasty (3 eyes),penetrating keratoplasty combined with cataract extraction (3 eyes),penetrating keratoplasty combined with trabeculectomy (2 eyes),and lamellar keratoplasty (1 eye).Conclusions Ultrasound biomicroscopy is important to guide the diagnosis and treatment of congenital corneal opacity.

Background Increase in ophthalmic optical medical instruments and microsurgical applications leads to retinal photochemical damage and other problems delivery of a variety of devices,so the in-depth study and understanding of its pathogenesis after retina light damage can provide a reference for the clinical treatment of related diseases.Objective This study was to investigate the therapeutic effect and relative mechanism of erythropoietin (EPO) on mouse retina photic injury by studying the expression of matrix metalloproteinases-2 (MMP-2)and MMP-9.Methods Fifty-two SPF BALB/c mice were randomized into normal control group,simple light-induced group and EPO pretreatment group by balloting method.The mice of simple light-induced group and EPO pretreatment group were continuously irradiated with 6000 lx diffuse light for 4 hours in a home-made box to establish the models of light-induced damage;while recombinant human EPO (rhEPO)of 5000 U/kg was intraperitoneally injected prior to the light exposure in the EPO pretreatment group.The expressions of MMP-2 and MMP-9 were examined at 6,12,36,72,96 hours and 7 days following light-exposure by immunohistochemistry.Results Edema and structural disorder of RPE cells appeared inthe simple light-induced group after light-exposure and aggravated with lapse of light-exposure time,but no similar change was seen until 7 days in the EPO pretreatment group.The immunohistochemistry findings showed that the expression of MMP-2(A value)in RPE cells was less in the normal mice.However,a large quantity of positive cells appeared in RPE layer 36 hours after light-exposure.Compared with the simple light-induced group,the positive expression of MMP-2 protein in EPO pretreatment group was significantly decreased,showing statistically significant differences among these three groups and different time points (Fgroup =3.68,P =0 04; Ftime =9.13,P=0.00).There was hardly any MMP-9 expression in the retina of the normal mice.In simple light-induced group,a few of positive cells appeared in RPE layer 6 hours after light-exposure and reached its peak 12 hours following light-exposure.The gradually down-regulation of MMP-9 expression happened 96 hours later following light-irradiation.The expression tendency of MMP-9 in EPO pretreatment group was similar to the simple light-induced group.Significant differences in expressions of MMP-9 were found among different groups and time points (Fgroup=3.61,P =0.04;Ftime =16.91,P=0.00).Conclusions MMP-2 and MMP-9 may be involved in the mechanism of retina photic injury by down-regulating the expression of MMP-2 and MMP-9 in RPE cells.

Objective To evaluate the clinical application of VAMTS in lung cancer patients.Meth- ods Between May 2004 and June 2005,30 patients with lung cancer applied VAMTS.The minimalthoracto- my from 8 to 10 cm was made at fourth or fifth intercostal space and lobectomy was undergone for convertion- al or dedicated endoscopic instruments.Some mediastinum lymphnode(MLN)resection were performed.Re- suits Operative time from 50 to 210 minutes and average 123 minutes.The intraoperative blood loss is less, and without blood transfusion.The average drawing duct time is 3.9 days.No death caused by operation and postoperative syndromes.Conclusion VAMTS wounds much less,operative scale lumination is better,opera- tive time shorter,blood loss less,and recover more quickly.The hospitalization time is shorter.The VAMTS with MLNR is regarded as an implement of lung cancer remedy,fits to stageⅠ-Ⅱ,diameter

Objective To delineate the potency of YC-1 on the proliferation,apoptosis,cell cycle and the protein expression of Caspase-3,-8,-9 in U937 and THP-1 leukemia cell lines.Methods MTT assay was performed to detect proliferation.Flow cytometry was used to measure the apoptosis and cell cycle.The expression of Caspase-3,-8 and-9 were detected by Western blot.Results The MTT assay showed that cell proliferation was inhibited in a concentration-dependent manner in 1.0,3.0,10.0 μmol/L YC-l-treated U937 and THP-1 cells.The survival rates for YC-1 after 24 h in U937 cells were (76.5±4.4) ％,(68.7±6.8) ％,(60.9±13.2) ％ respectively and (94.1±1.4) ％,(81.4±2.0) ％,(72.7±3.0) ％ respectively in THP-1 cell,compared with the control group (100 ％),there were significant differences (F =15.870,126.629,P ＜ 0.01).The apoptosis rates for 1.0,3.0,10.0 μmol/L YC-1 after 24 h were (40.7±1.0) ％,(55.6±2.3) ％,(71.8±1.5) ％respectively in U937 cells and (34.6±2.0) ％,(50.3±3.5) ％,(59.6±4.6) ％ respectively in THP-1 cells.With the control group (4.7±1.4) ％,(1.8±1.0) ％,there were significant difference (F =937.229,200.447,P ＜ 0.01).However,there was no significant difference for cell cycle.In addition,Cleaved Caspase-8 and Cleaved Caspase-3 expression after 1.0,3.0,10.0 μmol/L YC-1 treated for 24 h were significantly higher than control,but the expression of Caspase-9 did not appear significant change in U937 cells.As the same concentration and time point,Cleaved Caspase-3 expression increased with no change of Caspase-9 or Caspase-8 in THP-1 cells.Conclusion YC-1 effectively suppress the proliferation with little effect on cell cycle,but induce the apoptosis,have no effect on cell cycle,and the mechanism of apoptosis may be related to the Caspase activation in U937 and THP-1 cell lines.

Purpose:To study the effect of FasL gene on human lung adenocarcinoma cell lines and possibility of ex- ogenous FasL gene for gene therapy of lung cancer.Methods:An adenoviral expression vector with full length cDNA of FasL gene insert was constructed(Ad-FasL) and transfected into A549 cells.The effect of exogenous FasL gene on the growth of A549 cells was examined in vitro and in vivo.Results:Expression of FasL gene in A549 cells was confirmed by FCM and RT-PCR.The in vitro growth of the Ad-FasL transfected A549 cells was significantly inhibited (inhibition rate: 84%) as compared to mock (Ad-LacZ) transfected A549 cells.Colony-forming activity in vitro of the Ad-FasL transfected A549 cells was completely inhibited.The Ad-FasL transfected cells became apoptotic which was confirmed by the appear- ance of pre-G_1 on flow cytometry (FCM).The growth of A549 xenografts in nude mice was retarded by intra-tumol injection of Ad-FasL.Conclusions:FasL gene participates in the induction of cell apoptosis.Its use in gene therapy of cancer is promising.