Search

To evaluate the transfection efficiency of cultured chondrocytes from individuals with microtia (microtia chondrocytes) with the recombinant adeno-associated virus vector rAAV2-hVEGFsub165/sub-IRES-EGFP and hVEGFsub165/subin vitro. To test whether VEGFsub165/subgene-modified microtia chondrocytes can enhance the survival and quality of tissue-engineered cartilage.The recombinant plasmid rAAV2-hVEGF165-IRES-EGFP was inserted into rAAV2 virus vectors to construct rAAV2-hVEGF165-IRES-EGFP using the AATMaxTM system. The second-passage microtia chondrocytes were divided into 3 groups in vitro: the Ctr group (without transfection), Exp1 group (transfected with rAAV2-IRES-EGFP), and Exp2 group (transfected with rAAV2-hVEGF165-IRES-EGFP). At 24 h, 48 h, 72 h and 7 d after transfection, cell viability was measured by MTT staining. Transfection efficiency was determined by the rate of fluorescence-positive cells. The mRNA expression of hVEG165 was detected by RT-PCR (reverse transcription PCR) and agarose gel electrophoresis, and the VEGF165 protein levels in the supernatant fluids were measured by ELISAs. The second passage microtia chondrocytes with (Exp) and without (Ctr) transfection of VEGFsub165/subgenes were mixed with 0.5 ml 30% Pluronic F-127 at 4 °C and then injected subcutaneously into the opposing side of the back of nude mice. Eight weeks after injection, the cartilage-like tissues of nude mice were harvested for morphological and histologic examination.Chondrocyte viability increased in a time-dependent manner but did not differ among the 3 groups at the same time point. The mRNA and protein levels of VEGF increased in a time-dependent manner in the 3 groups. The mRNA and protein levels of VEGF165 were much higher in the Exp 2 group than in the Ctr and Exp 1 groups at the same time point, but the levels were not significantly different between the Exp 1 and Exp 2 groups. Both the Ctr group and the Exp1 group formed mature cartilage with mature lacunar structures, metachromatic matrices, collagen, and elastic fibers, and the structure of neonatal cartilage was not significantly different between the 2 groups. However, the wet weight of the neonatal cartilage was much larger in the Exp group (127.4 ± 12.4 mg) than in the Ctr group (58.5 ± 12.2 mg, plt; 0.05). VEGF protein staining also showed a higher level in the Exp group.The HVEGFsub165/subgene was transfected efficiently into microtia chondrocytes using the recombinant adeno-associated virus vector rAAV2-hVEGFsub165/sub-IRES-EGFP. After transfection, the mRNA and protein levels of hVEGFsub165/subincreased in a time-dependent manner. VEGFsub165/subgene-modified microtia chondrocytes showed enhanced survival in vivo but did not improve the texture of tissue-engineered cartilage.This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description ofthese Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Alleviating the pressure due to increasing freight transportation traffic by using low-emission and innovative transportation methods can reduce a number of problems, such as transportation...

Preventing environmental deterioration and alleviating traffic congestion are becoming urgent problems in urban and transportation planning. Alleviating the pressure from increasing freight transportation traffic via low-emission and innovative transportation methods can reduce problems such as transportation network capacity limits and environmental pollution and contribute to the development of resource-efficient and sustainable cities in the future. Underground freight transportation systems (UFTSs) can improve service quality and transportation efficiency in urban logistics and alleviate traffic congestion and associated problems such as energy consumption and air pollution. Previous studies on urban UFTSs have focused mostly on technical feasibility and policy requirements. Studies providing a quantitative analysis of the effects of introducing a UFTS on an existing transportation network are scarce. In this study, the main goal is to create a quantitative method to analyze the effects of introducing a UFTS on the performance of a transportation network. Thus, a multi-objective programming model of an integrated aboveground-underground transportation network that considers transportation cost, time, and emissions is created. The Yangshan port in Shanghai, China is used as an example to assess whether a UFTS can significantly reduce the cost, time, distance and emissions of the aboveground freight container transportation. The weights on three objective functions are varied to analyze their effects on the solution. These results provide a reference for optimizing freight distribution plans when a UFTS is constructed and for implementing integrated aboveground-underground transportation systems in the future.

In light of the rapid development of underground spaces in China and the intense demands of professionals for underground space, the Ministry of Education of the People’s Republic of China has permitted the offering of the urban underground space engineering professional degree program in universities since 2001. This professional degree has now been established in more than 80 universities across China. In this study, the authors have analyzed and discussed the background, history, disciplinary source, learning goals, curricula, and core courses of this profession in great depth. The authors hope that our findings shall contribute to the advancement and development of global efforts to train professionals in urban underground space engineering, promote the development of this profession, and improve the quality of professional cultivation efforts in this field, which in turn drives the scientific development and utilization of underground spaces.

Underground logistics systems (ULSs) has been an important topic and research hotspot for urban underground space use in China, especially for implementation in new cities and districts, as it is a necessary element to be considered for city underground space planning. Since these newly planned areas will face many challenges, including increasing logistics volume, spatial constraints etc. if we use the existing freight transport modes similar to old cities. Furthermore, the rapid developments of Chinese new cities provides an opportunity to implement innovative ULS freight transport systems and technology that are efficient and sustainable. Recently, ULS has been included officially in the planning and study for Beijing’s Sub-Center, the Yangtze River Demonstration District and other new cities and districts. However, the real value of ULS will not materialize unless it is successfully implemented. This paper presents the exploration, observations and findings of a few proposed ULS planning and research works in the new districts mentioned. The suitability of goods and freight types in ULS, logistic flow demand forecasting and ULS network systems integrated with underground metro or infrastructure are discussed. Both dedicated and integrated ULS network systems are also compared and analyzed. Despite the many advantages, many challenges and difficulties remain in the planning integration, technology adaptation and implementation of ULSs in new districts and cities, thus, interdisciplinary collaborations is one of the key success factors to ensure ULS can be brought to fruition.

In recent years, underground pedestrian system (UPS) has been developed worldwide, especially within central megacity areas, but research on UPS characteristics is still in the qualitative description stage, and quantitative methods are lacking to assess the impact of UPS on resilience. Therefore, it is essential to quantitatively measure UPS resilience and thus be able to make decisions on how UPS can be improved. The authors propose topological analysis of UPS networks based on computational and functional graph representations, which represents a new approach to theoretical discussions and empirical evidence for planning and configuring a UPS, thereby contributing to urban resilience. These representations provide functional views in which nodes represent single underground spaces and links represent underground sideways. Various graph measures are computed for structural analysis, and results provide insights into the required physical properties for planning resilient UPS. Based on validations applied to two typical high-density areas in Nanjing Xinjiekou and Guangzhou Zhujiang New Town, China, the authors examine how different UPS network patterns influence resilience performance, then further discuss the social and economic factors affecting network patterns, and give suggestions.

Astrocytes have been implicated in epileptogenesis and seizure-induced brain injury. Pathological studies reveal a variety of structural abnormalities in astrocytes, such as vacuolization and astrogliosis. While in vivo imaging methods have demonstrated rapid changes in astrocytes under a variety of physiological and pathological conditions, the acute effects of seizures on astrocyte morphology in vivo and corresponding mechanisms of seizure-induced astrocytic injury have not been documented. In this study, we utilized in vivo two-photon imaging to directly monitor the acute structural effects of kainate-induced seizures on cortical astrocytes. Kainate seizures cause an immediate, but transient, vacuolization of astrocytes, followed over several days by astrogliosis. These effects are prevented by pre- or post-treatment with rapamycin, indicating the mTOR pathway is involved in mediating seizure-induced astrocyte injury. These finding have clinical implications for mechanisms of seizure-induced astrocyte injury and potential therapeutic applications with mTOR inhibitors.

Summary The question mark ear is a malformation involving a cleft between the helix and the earlobe. The upper portion of the ear appears protruding and the scapha is absent from the affected region. The severity of this ear malformation varies from a small notch in the helix to complete separation of the helix and the lobe. In this article, we classify the question mark ear according to the severity of the malformation and propose two different methods of correction. In addition, we review the aetiology of the question mark ear and hope this can enhance the understanding of the pathology. Methods In the repair of moderate question mark ears, the local chondrocutaneous flap is used. In severe cases, tissue expander and autogenous rib cartilage are applied to reconstruct the deficiency of the lower part of the ear. Results There were 32 cases of question mark ears from July 2003 to December 2007. Thirty cases were sporadic and two cases had familial history. Twenty-two moderate question mark ears were repaired using chondrocutaneous flap transposition and ten severe question mark ears were repaired using tissue expander and autogenous cartilage. Conclusions In moderate question mark ears, the corrected auricle showed a shape similar to that of the unaffected auricle, but the size was a little smaller. Most of the reconstructed severe question mark ears showed excellent contour.

Urban underground transportation projects are introduced to address problems of scarce green land and traffic pollution. As construction of urban underground transportation is still in its infancy, there is no definite quantitative measurement on whether the construction is beneficial and what influences it will place on the region in China. This study intends to construct a comprehensive evaluation method for evaluating social, economic and environmental benefits of urban underground transportation projects and proposes the concept, role and principle for evaluation of environmental and economic benefits. It figures out relationship between the environment and factors of city development. It also summarizes three relevant factors, including transportation, biophysics and social economy, and works out indicators to evaluate the influence of urban underground transportation construction. Based on Contingent Valuation Method (CVM), Cost of Illness Approach (CIA), Human Capital Approach (HCA), this paper constructs 13 monetization calculation models for social, economic and environmental benefits in response to seven aspects, namely, reducing noise pollution and air pollution, using land efficiently, improving traffic safety, reducing traffic congestion, saving shipping time and minimizing transportation costs.

Objective Seizures cause acute structural changes in dendrites, which may contribute to cognitive deficits that occur in epilepsy patients. Disruption of the actin cytoskeleton of dendrites likely mediates the structural changes following seizures, but the upstream signaling mechanisms activated by synchronized physiological activity to cause seizure‐induced dendritic injury are not known. In this study, we test the hypothesis that the mechanistic target of rapamycin complex 1 (mTORC1) pathway triggers structural changes in dendrites in response to seizures. Methods In vivo multiphoton imaging was performed in transgenic mice expressing green fluorescent protein in cortical neurons. The effect of rapamycin pre‐ and posttreatment was tested on kainate‐induced dendritic injury and cofilin‐mediated actin depolymerization. Results Kainate‐induced seizures caused acute activation of mTORC1 activity, which was prevented by the mTORC1 inhibitor, rapamycin. Rapamycin pretreatment, and to a lesser degree, posttreatment attenuated acute seizure‐induced dendritic injury and correspondingly decreased LIM kinase and cofilin‐mediated depolymerization of actin. Interpretation The mTORC1 pathway mediates seizure‐induced dendritic injury via depolymerization of actin. These findings have important mechanistic and translational applications for management of seizure‐induced brain injury.

Posttraumatic epilepsy is a major source of disability following traumatic brain injury (TBI) and a common cause of medically-intractable epilepsy. Previous attempts to prevent the development of posttraumatic epilepsy with treatments administered immediately following TBI have failed. Recently, the mammalian target of rapamycin complex 1 (mTORC1) pathway has been implicated in mechanisms of epileptogenesis and the mTORC1 inhibitor, rapamycin, has been proposed to have antiepileptogenic effects in preventing some types of epilepsy. In this study, we have tested the hypothesis that rapamycin has antiepileptogenic actions in preventing the development of posttraumatic epilepsy in an animal model of TBI. A detailed characterization of posttraumatic epilepsy in the mouse controlled cortical impact model was first performed using continuous video-EEG monitoring for 16 weeks following TBI. Controlled cortical impact injury caused immediate hyperactivation of the mTORC1 pathway lasting at least one week, which was reversed by rapamycin treatment. Rapamycin decreased neuronal degeneration and mossy fiber sprouting, although the effect on mossy fiber sprouting was reversible after stopping rapamycin and did not directly correlate with inhibition of epileptogenesis. Most posttraumatic seizures occurred greater than 10 weeks after TBI, and rapamycin treatment for one month after TBI decreased the seizure frequency and rate of developing posttraumatic epilepsy during the entire 16 week monitoring session. These results suggest that rapamycin may represent a rational treatment for preventing posttraumatic epilepsy in patients with TBI.

Tuberous Sclerosis Complex (TSC) is a genetic disease involving dysregulation of the mTOR pathway and resulting in disabling neurological manifestations, such as epilepsy. Animal models may recapitulate epilepsy and other behavioral features of TSC and are useful tools for investigating mechanisms of epileptogenesis and other neurological deficits in TSC. In this chapter, methods for performing video-electroencephalography (video-EEG) to characterize epilepsy and neurological dysfunction in rodent models are reviewed. In particular, technical aspects of surgical implantation of EEG electrodes, video-EEG recording, and analysis and interpretation of EEG data are detailed. These methodological approaches should be helpful in characterizing seizures and background EEG abnormalities not only in animal models of TSC but also in many rodent epilepsy models in general.

Tuberous Sclerosis Complex (TSC) is a genetic disease involving dysregulation of the mTOR pathway and resulting in disabling neurological manifestations, such as epilepsy. Animal models may recapitulate epilepsy and other behavioral features of TSC and are useful tools for investigating mechanisms of epileptogenesis and other neurological deficits in TSC. In this chapter, methods for performing video-electroencephalography (video-EEG) to characterize epilepsy and neurological dysfunction in rodent models are reviewed. In particular, technical aspects of surgical implantation of EEG electrodes, video-EEG recording, and analysis and interpretation of EEG data are detailed. These methodological approaches should be helpful in characterizing seizures and background EEG abnormalities not only in animal models of TSC but also in many rodent epilepsy models in general.

The industry is development very quickly in P.R.China, it need logistics services provided by third party logistics (3PL) firms to reduce the costs. Nowdays,the technology and management push logistics knowledge development, so the logistics employee knowledge needs to be renovated to a higher lever. This paper analysed the industry firm services demand to 3PL firms based on the surveyed datum, it indicated that setting up multilayer logistics high education system and building qualification certification system are a good way to improve the logistics employee quality.

Summary Objective One of the greatest challenges in facial plastic surgery is total auricular reconstruction, as it demands precise technique as well as artistic creativity. The crucial step to reconstruct satisfactory auricles is sculpting a three-dimensional cartilaginous framework, which determines the success of surgery. In this paper we describe our new techniques in fabricating the three-dimensional auricular framework. Methods To assemble satisfactory auricular frameworks, we designed four different methods according to the number of rib cartilages used. (1) Method I, the framework was made from the entire 6th, 7th and 8th rib cartilages; (2) method II, the framework was made from the partial 6th, the entire 7th and 8th rib cartilages; (3) method III, the framework was made from the entire 7th and 8th rib cartilages; (4) method IV, the framework was made only from the entire 7th rib cartilage. Results 332 patients, ranging in age from 5 to 33 years old, were operated on using autogenous costal cartilage between 2004 and 2006. All the reconstructed ears had a satisfactory three-dimensional configuration. The cranioauricular angle of the reconstructed ears was also similar to that of the opposite ears. Conclusions According to the status of the normal ear and the development of the cartilage in each patient, we have developed four methods to harvest and fabricate cartilage. Our operations on 332 patients with microtia suggest that our new technique is effective in fabricating cartilaginous framework. Our new technique facilitates the operation procedure and minimises the usage of cartilage.

To investigate the feasibility of reconstruction of the contracted eye socket by an application of the expanded forehead island skin flap with the supratrochlear and supraorbital arteries.From June 2002 to June 2005, 6 patients with the eye socket defects were treated with an expanded forehead island skin flap with the supratrochlear and supraorbital arteries. There were 4 males and 2 females, aged 16-42 years. The defects were caused by tumors in 2 patients, by trauma in 3, and by chemical burns in 1; the defects were in the left eyes of 4 patients and in the right eyes of the remaining 2 patients, with the illness course of 1 year to 4 years. All the patients first underwent the skin and soft tissue expanding operation on the donor forehead skin area; 1 month later, the transplant of the expanded forehead island skin flap with the supratrochlear and supraorbital arteries was performed to reconstruct the eye sockets. The flaps ranged in size from 8 cm x 5 cm to 10 cm x 6 cm. The appearance and functional recovery of the reconstructed eye sockets were observed after operation.The follow-up of all the patients for 1-3 years revealed that the skin flaps survived, with no visible contracture, and the fine sensory function was still present. The artificial eyes could be steadily placed in the reconstructed eye sockets. The donor areas were healed with no visible hyperplastic scars.Reconstruction of the eye socket with an expanded forehead island skin flap with the supratrochlear and supraorbital arteries is a feasible, effective and simply method, and the patient can have a concealed incision, a satisfactory appearance, and a fine sensory function.

Ear reconstruction is one of the most challenging surgeries faced by the reconstructive surgeon. Currently, the use of autogenous costal cartilage is still best practice for microtia reconstruction. However, the relative deficiency of thin skin for coverage of the cartilage framework remains a limiting factor. Since 1994, we have used tissue expander in the mastoid area and innovated a technique to fabricate cartilage framework. All these innovations may solve the deficiency of the skin and improve cartilaginous framework definition. In conclusion, tissue expander offers a non-hairbearing, thin, well-vascularised skin to envelope an erect, contour-accentuated framework. Most patients with microtia are satisfied with their ear reconstruction.