To construct expression vectors of small hairpin RNA aimed at N-acetylglucosaminyltransferase Ⅴ(GnT-Ⅴ) gene, and to investigate effects of GnT-Ⅴ shRNA on proliferation, adhesion, migration and invasion of LoVo cell line. siRNAs were designed according to the coding sequence of GnT-Ⅴ gene, shRNA expression vectors were constructed and transfected into LoVo cell line, cell lines which stably expressed low level of GnT-Ⅴ were established by G418 screening. The mRNA and protein expression of GnT-Ⅴ were measured by semi- quantitative reverse transcription polymerase chain reaction(RT-PCR) and Western blot analysis, respectively. The effects of pGPU6/GFP/Neo GnT-Ⅴ shRNA vectors on proliferation, adhesion, migration and invasion of LoVo cell line were evaluated by CCK-8 assay, heterogenous adhesion, wound closure assay, chemotactic migration and cell invasive experiment, respectively. GnT- Ⅴ shRNA expression plasmid was constructed successfully and pGPU6/GFP/Neo GnT-Ⅴ shRNA down-regulated expression of GnT-Ⅴ dramatically in LoVo cell. Expression of LoVo GnT-Ⅴ/1564 and LoVo GnT-Ⅴ/2224 dereased by 82%, 71.5% respectively at mRNA level, and 68%, 56% respectively at protein level. The more effective interfered cell line, LoVo GnT-Ⅴ/1564, was chosen to do further experiment. CCK-8 assay showed proliferation of LoVo GnT- Ⅴ/1564 was suppressed obviously, compared to proliferation of negative control group cell (P

Acute Disease ; Adult ; Cholecystitis, Acute ; diagnosis ; pathology ; Diagnostic Errors ; Female ; Humans ; Myocarditis ; diagnosis ; pathology

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α(+) dendritic cells (DCs) in vitro was investigated in this study. Immature CD8α(+) DCs were prepared from C57BL/6 (H-2(b)) bone marrow cells by using a cytokine cocktail. On the 3rd day of culture, CD8α(+) DCs were pulsed by allogeneic (Balb/c, H-2(d)) EL9611 leukemia antigen, or RM-1 syngeneic prostate cancer antigen, with the concentration series of 0, 2.5, 5.0, 10.0, 20.0 μg/mL, respectively, then antigen-loaded immature CD8α(+) DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1:1, 2:1 and 4:1. T cell proliferation was measured by MTT assay. Cytokines including interferon gamma (IFN-γ) and interleukin-10 (IL-10) in CD8α(+) DCs and T co-culture supernatant were detected by using ELISA. Cytotoxic effect of antigen-specific T cells was tested by LDH release assay. Conventional mature DCs (mDCs) induced from C57BL/6 (H-2(b)) bone marrow cells by using granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control. The results showed that the proliferative activity of T cells stimulated by CD8α(+) DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α(+) DC/T ratio increased (P<0.05). When antigen concentration ≤ 5 μg/mL and CD8α(+) DC/T ratio ≤ 2:1, the ability of CD8α(+) DCs to stimulate T cell proliferation was higher than mDC control in allogeneic tumor antigen-pulsed groups (P<0.05), but not in syngeneic tumor antigen-pulsed groups (P>0.05). The level of IFN-γ and IL-10 in CD8α(+) DCs and T cell co-culture supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P<0.05), and the cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when the CD8α(+) DC/T was 1:1 or 2:1 (P<0.05). There existed a negative correlation between the level of IL-10 and T cell proliferation. T cell cytotoxicity assay showed that when CD8α(+) DCs were pulsed with allogeneic tumor antigen, the maximal T cell killing efficiency could reach (100±7.7)%, whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)%. It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α(+) DCs could stimulate T cells to exert the GVT effect in vitro, and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen. The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α(+) DC/T ratio (1:1 and 2:1).

Background There is no effective method to regenerate the optic nerve after injury. It has been recently reported that α-crystallin could promote the survive rate and axon regeneration of retinal ganglion cells (RGCs) effectively. However,the molecular mechanism is not clear. Objective This study was to identify the site of RGCs where the exogenous α-crystallin bind to. Methods RGCs was isolated from retinas of two 2-day-old Long Evans rats and primarily cultured. The positive rate of the RGCs was assessed by counting the number of positive cells for fluorescently-labeled thy1. 1 and cy3 under the fluorescence microscope. The biotinylated exogenous α-crystallin was evaluated by direct coloration and the activity of molecular chaperones was measured by insulin test.After identifying the success of biotinylation along with the activity of molecular chaperones,biotinylated α-crystallin was co-incubated with RGCs and the cells then were reacted to fluorescently labeled avidin for the observation of binding site of exogenous α-crystallin under the laser confocal microscope. Results RGCs of 94% were survived through primary culture. The coloration of biotinylated α-crystallin labeled by the direct coloration method was more intensive, and the value of A450 descended as the decrease of biotinylated α-crystallin concentration,indicating that the α-crystallin was biotinylated successfully. The activity of molecular chaperones of biotinylated α-crystallin was significantly strong but no significant change after being biotinylated after co-incubation of RGCs with biotinylated α-crystallin. Laser confocal microscope examination revealed that co-incubated RGCs with biotinylated α-crystallin showed the red fluorescence on membrane and axon of RGCs rather than cytoplasm and nucleus. The absent response was seen in the control group. Conclusion Exogenous α-crystallin can specifically combine with the membrane of RGCs to play the biological function,but its binding mode and mechanism need further study.

Abstract?AlM:To investigate the choice of different treatments for malignant glaucoma.? METHODS: ln this retrospective case series, 21 malignant glaucoma patients ( 21 eyes ) admitted in Wuhan General Hospital of Guangzhou Military Command from May 2012 to May 2013 were analyzed. Sixteen eyes ( 76%) developed malignant glaucoma after filtration surgery, 3 eyes ( 14%) after EX - PRESS glaucoma filtration device, 2 eyes ( 10%) after glaucoma filtration Ahmed valve implantation. Main Outcome of corrected visual acuity, intraocular pressure ( lOP ) , anterior chamber depth and complications were detected.?RESULTS: lOP recovered by drug control in 13 eyes, anterior chamber depth. Four eyes were treated by vitreous water- bag aspiration combined with anterior chambers reconstructing. Two eyes were treated by cataract extraction and intraocular lens implantation. Two eyes were treated by posterior capsule excision combined with anterior vitrectomy. lOP before and after treatment was 29. 81±4. 98, 12. 71±3. 77mmHg, respectively (P=0. 00). Anterior chamber depth before and after treatment was 0.41± 0. 34, 2. 13 ± 0. 54mm, respectively (P = 0. 00). Corrected visual acuity before treatment was 0. 19 ± 0. 17, after treatment was 0. 20±0. 16 (P= 0. 36). Except for vitreous hemorrhage in 1 eye, there were no ocular or systemic adverse events observed in all patients.? CONCLUSlON: lt is good to diagnose malignant glaucoma in early period, and treated it step by step. For this can reduce lOP and restore anterior chamber.

Objective To observe the protective effects of melatonin against spinal cord injury from seawater immersion in rabbits.Methods The 120 mature and health New Zealand White rabbits,weight range from 2.6 to 2.9kg,were randomly divided into four groups (30 each):control group,ethanol group,melatonin group (100mg/kg),methylPrednisolone group (30mg/kg).The rabbit model of spinal cord injury were built by modified Allen's method taking the 10th thoracic vertebra as a center,seawater immersion for 60 minutes,and then by grouping to give the appropriate treatment.After each group was given the corresponding treatment,six rabbits in each group were randomly selected at 1,6,12,24 and 48 hours five different time points.The neurological function scores of the rabbits were evaluated by Tarlov method,the spinal cord ofT9 to T.which were obtained from all the groups were used for further study,including immunohistochemical detection of apoptosis proteins:Bax,Bcl-2,neurofilament protein 200 (NF200) and in situ end labeling (TUNEL) method to detect spinal neuronal cell apoptosis.Results Within each observation time point,the Tarlov score was higher in melatonin group and methylprednisolone group compared with control group and ethanol group (P＜0.05),there was no significant difference between melatonin group and methylprednisolone group (P＞0.05).The expressions of Bcl-2 and NF200 were significantly higher in melatonin group and methylprednisolone group compared with control group and ethanol group,while Bax expression was significantly lower (P＜0.05).There were no significant difference between melatonin group and methylprednisolone group in the expression of three proteins (P＞0.05).The TUNEL-positive apoptotic cells were fewer in melatonin group and methylprednisolone group compared with control group and ethanol group (P＜0.05),and there was no significant difference between melatonin group and methylprednisolone group (P＞0.05).Conclusion Melatonin has protective effect against spinal cord injury from seawater immersion in rabbits,no difference in efficacy exists compare with methylprednisolone.

OBJECTIVE:To observe clinical efficacy of bevacizumab or cetuxizumab combined with FOLFOX4 regimen in the treatment of advanced rectal cancer. METHODS:114 patients with rectal cancer were randomly assigned to cetuxizumab group and bevacizumab group,with 57 cases in each group,among which one patient of bevacizumab group withdrew from therapy. Both groups received FOLFOX4 regimen:oxaliplatin 85 mg/m2+calcium folinate 200 mg/m2,ivgtt,2 h,and 5-FU 400 mg/m2,ivgtt, last,5-FU 600 mg/m2,ivgtt,22 h. Cetuxizumab group was additional given cetuxizumab 500 mg/m2;bevacizumab group was addi-tionally given bevacizumab 5 mg/kg,ivgtt. A treatment course lasted for 2 weeks. Both groups received 4 courses of treatment,and then clinical efficacy,toxic reaction and progression-free survival (PFS) were evaluated. RESULTS:Objective remission rate (RR),disease control rate(DCR)and median PFS of cetuxizumab group was 45.61%,92.98%and 10.0 months,those of bevaci-zumab group were 48.21%,87.50%and 11.0 months;there was no statistical significance between 2 groups(P>0.05). No signifi-cant differences were found in the incidence of ADR such as sensory neurotoxicity,aleucocytosis,thrombopenia,nausea and vomit-ing,diarrhea and erythra between 2 groups(P>0.05). CONCLUSIONS:Both bevacizumab or cetuxizumab combined with FOLF-OX4 regimen have a similar effect on patients with advanced cancer,with low incidence of toxic reaction.

AIM: To investigate the incidence of trachoma in children aged 1 to 9y in Hainan Province and determine high-risk trachoma endemic and non-endemic areas in Hainan, and thus provide evidence for developing trachoma control and prevention therapy.METHODS:The areas of investigation were chosen on the basis of past literatures, expert interviews and survey on the spot.In 2013, Hainan Provincial Office of Blindness Prevention carried out the survey in 7 counties including Dongfang City, Wuzhishan City, Ledong County, Baisha County, Baoting County, Lingao County and Changjiang County.In these districts, 356 pupils including 192 boys and 164 girls were examined, their age ranging from 1 to 9 and their average age being 7 years old.The targeted students received the trachoma rapid assessment by the adoption of simplified trachoma classification system which was recommended by the World Health Organization.RESULTS: No case of active trachoma was found among the 356 students.CONCLUSION:The prevalence rate of trachoma in children under 9 years is less than 5% in Hainan Province.Active trachoma is not a public health issue in Hainan Province.

AIM: To investigate the safety and effect of EX-PRESS glaucoma drainage device on open-angle glaucoma.
METHODS: A retrospective review of 40 patients ( 47 eyes ) whose eyes were diagnosed as open angle glaucoma through the best - corrected visual acuity ( BCVA ) , intraocular pressure ( IOP ) , gonioscope, examination of the ocular fundus, and visual fields et al. Fourty patients 6-73y old ( mean 39. 90±16. 50y old) with the BCVA from 0. 01 to 0. 6 and the IOP from 18mmHg to 65mmHg were treated with EX-PRESS glaucoma drainage device after achieving not well IOP over three drugs. Follow - up of these patients 1d, 3d and 1wk after treatment, IOP, slit lamp examination were retrospectively observed. The changes of the BCVA, IOP were used to evaluate the safety and effect before and after treatmeat.
RESULTS: The mean BCVA was 0. 26 ± 0. 29 pre -operatively, while it was 0. 24±0. 22 after treatment one week. The mean BCVA decreased slightly, but there was no statistic difference between pre-treatment and post-treatment (t=1. 56,P=0. 13). The mean IOP was (36. 62 ±14.01)mmHg pre-operatively, (10.04±5.77)mmHg after treatment one day , ( 9. 59±4. 93 ) mmHg after treatment three days, (9. 47±3. 06)mmHg after treatment one week. The IOP was decreased significantly in post-treatment after one day, three days, one week compared with pre-treatment (F=157. 20, P<0. 05). Except for 5 eyes of the IOP below 5mmHg, there was no ocular or systemic adverse events observed in all patients.
CONCLUSION:EX-PRESS glaucoma drainage device is an effective and safe treatment for the patients with open-angle glaucoma. The risk and complication are low intraoperatively and postoperatively.