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.

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.

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 analyze the incidence and risk factors for regional nodal failure(RNF) and chest wall recurrence(CWR) in T1 or T2 breast cancer patients(median age 44 year-range 26-72) with 1-3 positive axillary nodes treated with postmastectomy radiotherapy limited to the regional nodes.Methods From 1990 to 1999,320 patients were treated with postmastectomy(radical or modified radical) radiother- apy confined to the supraclavicular and internal mammary nodes with a median dose of 50 Gy in 25 fractions over 5 weeks.The median number of nodes examined was 8 (range 1-24).The median lymph nods rate (LNR) was 25% (range 5%-100%).Results The 5-year overall survival rate and disease free survival rate was 89.7% and 83.4%,respectively.The 5-year RNF and CWR was 7.9% and 5.7%,respectively. The 5-year RNF in patients with LNR＜30% and≥30% was 4.4% and 14.0% (P=0.002).The 5-year CWR in the subgroups with LNR＜30% and≥30% was 3.5% and 9.6% (P=0.018).In age≤35 year eld patients with LNR≥30%,the 5-year RNF and CWR was 40.0% and 20.0%.In T2 patients with LNR≥30%,the 5-year RNF and CWR was 15.8% and 12.2%.Age and LNR were independent prognostic factors for RNF+CWR,LNR was the only independent prognostic factor for CWR by multivariate analysis. Conclusions In T1 or T2 breast cancer patients with 1-3 positive axiliary nodes treated with radical or modified radical mastectomy,a relatively high incidence of chest wall recurrence is observed in the subgroup of patients with lymph nods rate of 30% or greater accompanied by a T2 primary tumor or age≤35 years old. Lymph nodes rate is the only significant prognostic factor of chest wall recurrence.For these patients,post- operative lymphatic drainage area and chest wall irradiation should be considered.

Objective To isolate and purify Thy 1 low Lin Sca 1 + bone marrow stem subset cells by magnetic activated cell sorting(MACS). Methods Thy 1 low Lin Sca 1 + cells from mouse bone marrow were collected through three processes by MACS.After Lin + cells were removed,Thy 1 low Lin Sca 1 + bone marrow stem cell subsets were harvasted.The purity of the cells was analysed by FACS and the reclaimation rate was counted. Results The purity and reclaimation rate of Thy 1 low Lin Sca 1 + cells were 72.36% and 82.43% respectively, which equaled to the level of isolation and purification of CD34 + cells by MACS. Conclusions It is effective to isolate and purify Thy 1 low Lin Sca 1 + bone marrow stem cell subsets by MACS, and the purity and reclaimation rate of the cells are high.

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.

Objective To evaluate the clinical application of CT-guided localization with combination of methylene blue and a Hookwire system for small pulmonary nodules (SPNs) before video-assisted thoracoscopic resection.Methods CTguided localization the SPNs before resection in 56 patients and 60 nodules,then underwent video-assisted thoracic surgery (VATS) resection.Among 56 patients,19 males and 37 females,aged from 35 to 81 years,mean age was (61.1 ±8.9)years.Results SPNs diameter (6.80 ±4.12) mm,distance from the parietal pleura (15.38 ±4.63) mm.CT-guided localization success rate was 100％,positioning time (10.76 ± 8.17) min,8.9％ (5/56) had micro pneumothorax aftet positioning,7.1％ (4/56) occurrence of needle tract bleeding,no conservative treatment.VATS resection rate was 100％.The pathology of 60 lesions were shown:Bronchiolo-alveolar carcinoma(BAC) were 33 lesions(55.0％),BAC and adenocarcinoma were 11 lesions(18.3％),Atypicaladenomatous hyperplasia (AAH) were 7 lesions (11.8％),Inflammation were 4 lesions (6.7％),Harmatoma were 3 lesions(5.0％),Tuberculoses were 2 lesions(3.3％).Conclusion CT-guided localization with combination of methylene blue and a Hookwire system before video-assisted thoracoscopic resection is a promising technique for small solitary pulmonary nodules.It could play an important role in accurate localization of small pulmonary nodules,and it is a safe technique with clinical application.