Age-related macular degeneration (AMD) has become one of the leading causes of irreversible blindness worldwide.With the advancement of stem cell technology,tissue engineering and biomaterials,cell-based therapy has been inspiring for many degenerative diseases.For its unique advantages,AMD has become one of the most promising fields for cell-based therapy,which involve retinal pigment epithelium(RPE)cells,induced differentiation of neural retina cells and related cytokine regulations.RPE cells can be derived from human embryonic stem cells (hESC) or Induced pluripotent stem cells (iPS).Recently hESC-derived RPE cells have been applied to patients with dry AMD with initial success in clinical trials.In terms of tissue engineering,studies are focused on factors affecting the long-term survival of transplanted cells,including tissue scaffolds,soluble hybrid materials and scaffold anchoring.This article briefly reviews the RPE differentiation,neural retina differentiation and related cytokines of cell-based therapy and scaffolds,materials,and cell-scaffolds interactions of tissue engineering in AMD treatment.

Objective To analyze the aetiological characteristics and bacterial susceptibility in chronic obstructive pulmonary disease (COPD) complicated with ventilator-associated pneumonia (VAP) after mechanical ventilation. Methods To review the clinical characteristics, aetiological flora and bacterial susceptibility in 28 patients complicated with VAP in the ICU in recent years. Results The incidence rate of VAP was 66.7％. 91 clinical bacteria and 10 mycete isolates were collected, 64 (80.3％) were Gram-negative bacteria (GNB) and 27(29.7％) Gram- positive bacteria (GPC). The dominant bacteria was Xanthomonas maltophilia in COPD complicated VAP. Conclusion The main kind of bacteria in COPD complicated VAP after mechanical ventilation was GNB, and all kinds of the bacteria showed high tolerance. More attention should be paid to the cultivate aetiological bacteria and bacterial susceptibility and select the most suitable antibiotics in the treatment of VAP.

Objective:To review effects of functional electrical stimulation(FES) on paralyzed or paretic muscles,and introduce the progress of functional electrical stimulation repairing spinal cord injury.Methods:A computer-based search was conducted in CNK/and MEDILINE for articles related to functional electrical stimulation repairing spinal cord injury from November 1990 to November 2009 with the Keywords of spinal cord injury and electrical stimulation.Data were check firstly.Articles related to functional electrical stimulation repairing spinal cord injury were selected and looked for full-texts.The articles were analyzed and summarized.Results:Functional electrical stimulation (FES) can activate paralyzed or paretic muscles to generate functional or therapeutic movements with constant frequency.FES can improve the impaired motor function of muscles stimulated.Functional electrical stimulation technique could repair part of motor function following spinal cord injury.Conclusions:Functional electrical stimulation is a new and promising technique in modem rehabilitation engineering.Functional electrical stimulation (FES) can activate paralyzed or paretic muscles to generate functional or therapeutic movements.FES can improve the impaired motor function of muscles stimulated.

Aim: To clone and over-express the gene encoding aminoglycoside(AG)phosphotransferase(APH)from clinical MRSA isolates in E.coli and to develop an assay method for the recombinant APH.Methods: The susceptibility of clinical MRSA isolates to AGs was tested by disk diffusion.A nucleic acid sequence encoding APH was amplified from the genomic DNA of an isolate and ligated to expression vector pET-28a,and then trans-formed into E.coli BL21(DE3).After purification of the recombinant protein by affinity chromatography,the phosphorylation activity of the enzyme was determined by ESI-MS and disk diffusion.Flesults: All 6 clinical MRSA isolates were unsusceptible to AGs.After cloning and expression,the recombinant APH was purified to90%.The in vitro activity assay indicated that the recombinant protein could inactivate kanamycin B in the assay mixture within 2 h.Conclusion: The recombinant APH showed excellent enzymatic activity.The assay method was simple and convenient,which may provide the basis of developing a screening model for APH inhibitors.

Objective To determine if pregnancy affects the toxicity of bupivacaine and to investigate the effect of Shenfu injectio,a preparation of Chinese herbal medicine,on central nervous system and cardiac toxicity of bupivacaine in pregnant rats.Methods Twenty-four SD rats weighing 320-360 g were assigned to 3 groups(n =8 each):Ⅰ non-pregnant control group,Ⅱ pregnant control group and Ⅲ Shenfu injectio pretreatment group. The animals were anesthetized with isoflorane(2%-4%)-O_2 inhalation which was stopped before bupivacaine infusion was started.Femoral artery was canunlated for MAP monitoring and blood sampling and femoral vein was cannulated for bupivacaine infusion.MAP,HR and ECG were continuously monitored.All animals in the 3 groups received continuous infusion of 5% bupivacaine at 2 mg?kg~(-1).min~(-1).In group Ⅲ Shenfu injectio 10 ml?kg~(-1) was injected intraperitoneally(IP)30 min before bupivacaine infusion whereas in the two control groups(group Ⅰ and Ⅱ)equal volume of normal saline was injected IP instead of Shenfu injectio.The duration between the beginning of bupivacaine infusion and onset of convulsion/arrhythmia(QRS≥90 ms)/asystol was recorded and the amount of bupivacaine infused was calculated.Results There was no significant difference in the amount of bupivacaine causing convulsion and asystol between group Ⅰ and Ⅱ but the amount of bupivacaine causing arrhythmia was significantly larger in group Ⅰ(non-pregnant) than in group Ⅱ(pregnant control group)(P＜0.05).The amount of bupivacaine causing convulsion,arrhythmia and asystole was significantly larger in Shenfu injectio pretreatment group(group Ⅲ)than in pregnant control group(group Ⅱ)(P＜0.05 or 0.01).Conclusion Bupivacaine- induced cardiotoxicity is increased in pregnant rats and Shenfu injectio pretreatment can reduce the systemic toxicity of bupivacaine in pregnant rats.

Epstein-Barr Virus Infections ; Herpesvirus 4, Human ; genetics ; Humans ; MicroRNAs ; Neoplasms ; virology

BACKGROUND:Intertrochanteric fracture is one of the most common fractures in older adults. The surgical treatment methods include artificial joint replacement (total hip and femoral head replacement) and internal fixation (proximal femoral nail anti-rotation and dynamic hip screw), but there is no clear evidence to evaluate the effect of them. OBJECTIVE: To compare the effect difference of arthroplasty and internal fixation in the treatment of intertrochanteric fracture in the elderly people. METHODS:We retrieved randomized controled studies on artificial joint replacement (total hip and femoral head replacement) and internal fixation (proximal femoral nail anti-rotation and dynamic hip screw) in the treatment of intertrochanteric fracture from 1990 to 2015. Meta-analysis was used to compare operation time, intraoperative bleeding volume, one-year postoperative Harris hip scores, and one-year postoperative complication between the arthroplasty and internal fixation. RESULTS AND CONCLUSION:(1) Twenty-one studies were included. (2) Intraoperative bleeding volume and one-year postoperative Harris hip scores were higher in the arthroplasty group than in the internal fixation group. (3) Operation time was similar between the arthroplasty and internal fixation groups. (4) Complication rate was lower in the arthroplasty group than in the internal fixation group. Incidence of complications was closed between the arthroplasty and internal fixation groups. (5) Results suggested that the effect of arthroplasty was better than internal fixation for treating intertrochanteric fracture in the elderly. However, prospective large-sample long-term randomized controled trials are needed for verification.

Background The establishment of diabetic animal model is a crucial step for the study about diabetic eye diseases. At present,the main modeling method include the injection of streptozotocin and alloxan. But the shortcoming of the former is an expensive price, and that of the later is high death rate of animals. Objective This experiment was to discuss the way which decrease the death of alloxan-injected animal and explore the effects of high blood glucose on the posterior capsular opacification (PCO). Methods Forty clean healthy male New Zealand white rabbits were randomly divided into 2 groups. 90mg/kg of alloxan were injected via ear vein once in 20 rabbits to create the diabetic animal models,and the equivalent amount of normal saline solution was injected at the same way as normal blood glucose group. The successful models were selected in the animals with the blood glucose level over 12. 0 mmol/L two weeks later, and PCO of lens were graded based on the method of Odrieh under the slit lamp. Extracapsular lens extraction was then performed on the right eye of rabbits in both groups, and the posterior capsules were obtained from these eyes at the 6th, 10th and 14th days after operation. The expression of proliferating cell nuclear antigen ( PCNA ) in posterior capsular lens epithelial cell was detected by immunohistochemistry. Results The modeling successful rate was 70% after injection of alloxan. The body weight of rabbits in high blood glucose group was significantly lowed and the blood glucose was significantly elevated in comparison with normal blood glucose group ( all P<0. 05). Two weeks after surgery ,2 eyes occurred 2 grade of PCO and only one eye showed the 1 grade of PCO in the high blood glucose group. However, 1 grade of PCO was found in 3 eyes in the normal blood glucose group. Biopsy revealed that PCNA was positively expressed in the cell nuclei of LECs in high blood glucose group rather than the normal blood glucose group from the 10th day after surgery. The proliferation index of PCNA was 0. 86±0. 04 and 0. 25±0. 03 respectively in high blood glucose group and normal blood glucose group, showing a significant difference between them (t = -16. 171 ,P = 0. 000). Conclusion Stable diabetic models of rabbits can be created by intravenous injection of 90 mg/kg alloxan. High blood glucose level is one of the important factors for the development of PCO.

This study examined the current changes of human ether-a-go-go-related gene (hERG) mutation derived from a LQT2 Chinese family with a highly penetrating phenotype. Mutation was identified and site-directed mutagenesis was performed to induce the mutation in wild-type (WT) hERG. WT hERG and mutated V535M were cloned and transiently expressed in HEK293 cells. At the 48th and 72nd h after transfection, membrane currents were recorded using whole cell patch-clamp procedures. An A>G transition at 1605 resulting in replacement of V535M was identified. Compared to WT, V535M mutation significantly decreased tail currents of hERG. At test potential of -40 mV after depolarizing at +50 mV, tail current densities were 83.35±7.06 pA/pF in WT and 50.38±7.74 pA/pF in V535M respectively (n=20, P<0.01). Gating kinetics of hERG revealed that V (1/2) of steady-state inactivation shifted to negative potential in the mutant (V (1/2,V535M): -61.81±1.7 mV vs. V (1/2, WT): -43.1±0.71 mV). The time constant of recovery from inactivation was markedly prolonged in the mutant compared to WT among test potentials. V535M hERG mutation demonstrated markedly decreased tail current densities, which suggests that V535M is a new loss-of-function mutation of hERG channel responsible for LQT2.

Background Growth factor-induced proliferation and migration of retinal pigment epithelium (RPE) cells are the major pathological changes of proliferative vitreoretinopathy (PVR).Arsenic trioxide ( As2O3 ) is an active ingredient of Chinese traditional medicines,which has an inhibition on proliferation and migration of tumor cells.However,it is not clear whether As2O3 could inhibit growth factor-induced proliferation and migration of RPE cells. Objective This study was to explore the effects of As2O3 on epidermal growth factor (EGF)-induced proliferation and migration of ARPE-19 cells. Methods RPE cell line (ARPE-19 cells) were cultured.Different concentrations of As2O3(0,0.5,1.0,2.0,5.0,10.0,20.0 μmol/L) were added in the culture plate to treat ARPE-19 cells with or without 10 mg/L EGF in serum-free group for 24 and 48 hours,respectively.The MTT colorimetric assay was used to check the cell viability and evaluate the drug toxicity.The effects of As2O3 on EGF-induced proliferation of ARPE-19 cells were analyzed to get an effective and avirulent concentrations of As2O3.The effects of As2O3 on EGF-induced migration of ARPE-19 cells were observed by scratch-wound assay and the Boyden chamber assay.Results MTT assay showed that the A values were gradually declined with the increase of As2O3 concentrations after As2O3 treatment without EGF for 24 hours and 48 hours ( Fgroup =38.269,P =0.000 ; Ftime =0.874,P =0.358 ).Compared with the control group,no significant differences were seen in the A values of ARPE-19 cells in 0.5-5.0 μmol/L groups (all P＞0.05).Meantime,As2O3 reduced the A values of ARPE-19 cell with 10 mg/L EGF in dose- and time-dependent manner ( Fgroup =152.155,P =0.000 ; Ftime =51.649,P =0.000 ).There were not significant differences in 10 mg/L EGF-induced cell growth after 0.5,1.0,2.0 μmol/L As2O3 was added for 24 and 48 hours ( Fgroup =2.215,P =0.126 ;Ftime =2.230,P =0.155).However,when 5.0-20.0 μmol/L As2O3 added,the A values of 10 mg/L EGF-induced ARPE-19 cells lowed,showing a significant difference in comparison with the control groups ( all P＜0.05),with the cellular inhibiting rate 12％,32％,37％ in 24 hours and 39％,44％ and 53％ in 48 hours.Scratch-wound assay showed that EGF-induced horizontal migration of ARPE-19 cells was slow after 0.5-2.0 μmol/L As2O3 treated,and the same results also appeared in cell lognitudinal migration by Boyden chamber assay,with the inhibitory rates 22％,33％ and 46％ respectively. Conclusions As2O3 is avirulent on ARPE-19 cells within definite concentration range.At ≤ 2.0 μmol/L concentrations,As2O3 dose not affect EGF-induced proliferation of ARPE-19 cells,but it suppresses EGF-induced cell migration.At ≥ 5.0 μmol/L concentrations,As2O3 plays an inhibitory role to EGF-induced proliferation of ARPE-19 cells.