The Effect of Hemodynamic Remodeling on the Survival of Arterialized Venous Flaps

Objective

To evaluate the effect of hemodynamic remodeling on the survival status of the arterialized venous flaps (AVFs) and investigate the mechanism of this procedure.

Materials and Methods

Two 7 x 9 cm skin flaps in each rabbit (n=36) were designed symmetrically in the abdomen. The thoracoepigastric pedicle and one femoral artery were used as vascular sources. Four groups were included: Composite skin grafts group and arterial perfusion group were designed in one rabbit; AVF group and hemodynamic remodeling group by ligation of the thoracoepigastric vein in the middle were outlined in another rabbit. Flap viability, status of vascular perfusion and microvasculature, levels of epidermal metabolite and water content in each group were assessed.

Results

Highly congested veins and simple trunk veins were found using angiography in the AVF group; while a fairly uniform staining and plenty of small vessels were observed in the hemodynamic remodeling group. The metabolite levels of the remodeling group are comparable with those in the arterial perfusion group. There was no statistically significant difference in the percentage of flap survival between the arterial perfusion group and hemodynamic remodeling group; however, significant difference was seen between the AVF group and the hemodynamic remodeling group.

Conclusions

Under the integrated perfusion mode, the AVFs are in an over-perfusion and non-physiological hemodynamic state, resulting in unreliability and unpredictability in flap survival; under the separated perfusion mode produced by remodeling, a physiological-like circulation will be created and therefore, better flap survival can be expected.     

Rapid Genetic Analysis of Epithelial-Mesenchymal Signaling During Hair Regeneration

Hair follicle morphogenesis, a complex process requiring interaction between epithelia-derived keratinocytes and the underlying mesenchyme, is an attractive model system to study organ development and tissue-specific signaling. Although hair follicle development is genetically tractable, fast and reproducible analysis of factors essential for this process remains a challenge. Here we describe a procedure to generate targeted overexpression or shRNA-mediated knockdown of factors using lentivirus in a tissue-specific manner. Using a modified version of a hair regeneration model ^{5, 6, 11}, we can achieve robust gain- or loss-of-function analysis in primary mouse keratinocytes or dermal cells to facilitate study of epithelial-mesenchymal signaling pathways that lead to hair follicle morphogenesis. We describe how to isolate fresh primary mouse keratinocytes and dermal cells, which contain dermal papilla cells and their precursors, deliver lentivirus containing either shRNA or cDNA to one of the cell populations, and combine the cells to generate fully formed hair follicles on the backs of nude mice. This approach allows analysis of tissue-specific factors required to generate hair follicles within three weeks and provides a fast and convenient companion to existing genetic models.     

响应面法对小球藻Chlorella zofingiensis高产虾青素条件的优化

采用析因设计法(Plackett-burman)对影响Chlorella zofingiensis高产虾青素的相关因素进行评价,发现硝酸钠、光照强度、二价铁离子及醋酸钠浓度对虾青素产量影响显著.利用中心组合设计(central composite design)及响应面分析对影响虾青素产量的关键因素做进一步的优化,得到较佳的试验点为二价铁离子浓度0.41 mmol/L,硝酸钠浓度0.8 mmol/L,醋酸钠浓度37.1 mmol/L,光照强度650 E/m2×s.优化后虾青素产量从7.890mg/L提高到19.81mg/L,比优化前提高了2.5倍.     

Immobilization of Rhizopus oryzae in a modified polyvinyl alcohol gel for L(+)-lactic acid production

The production of L(+)-lactic acid (LA) by Rhizopus oryzae immobilized in polyvinyl alcohol (PVA) was investigated. To decrease diffusional resistance, we modified the PVA gel through the addition of sodium alginate and phosphate esterification. The production of L(+)-LA improved notably in the immobilized Rhizopus oryzae. Maximum L(+)-LA production (106.27 g/L), with a yield of 73.1 % and rate of 2.95 g/L·h, was obtained at a temperature of 38 °C, 6 % PVA, and 0.8 % sodium alginate. The immobilized R. oryzae was stable in 14 serial-batch cultures using non-growth medium. The immobilized beads also displayed good tolerance to low temperature and long-term storage at 4 °C with the preservation of biochemical properties.     

Sandwich‐Type Microporous Carbon Nanosheets for Enhanced Supercapacitor Performance

Sandwich‐type microporous hybrid carbon nanosheets (MHCN) consisting of graphene and microporous carbon layers are fabricated using graphene oxides as shape‐directing agent and the in‐situ formed poly(benzoxazine‐co‐resol) as carbon precursor. The reaction and condensation can be readily completed within 45 min. The obtained MHCN has a high density of accessible micropores that reside in the porous carbon with controlled thickness (e.g., 17 nm), a high surface area of 1293 m² g^?1 and a narrow pore size distribution of ca. 0.8 nm. These features allow an easy access, a rapid diffusion and a high loading of charged ions, which outperform the diffusion rate in bulk carbon and are highly efficient for an increased double‐layer capacitance. Meanwhile, the uniform graphene percolating in the interconnected MHCN forms the bulk conductive networks and their electrical conductivity can be up to 120 S m^?1 at the graphene percolation threshold of 2.0 wt.%. The best‐practice two‐electrode test demonstrates that the MHCN show a gravimetric capacitance of high up to 103 F g^?1 and a good energy density of ca. 22.4 Wh kg^?1 at a high current density of 5 A g^?1. These advanced properties ensure the MHCN a great promise as an electrode material for supercapacitors.     

敲除Nrf2促进高脂饮食诱导小鼠肝脏NF-kB的活化

目的：氧化应激和炎症反应是NASH进展的关键因素,同时二者之间存在着密切关系,而转录因子Nrf2和NF-kB分别是氧化应激和炎症信号通路的关键调控靶点,因此,研究Nrf2对高脂饮食诱导小鼠肝脏NF-kB信号通路的影响,对探讨NASH进展具有重要的意义。方法：雄性野生型（WT）和Nrf2基因敲除（Nrf2-/-）ICR小鼠各10只,随机分为WT对照组（Control）、Nrf2-/-对照组（KO）、WT高脂饮食组（HFD）和Nrf2-/-高脂饮食组（KOHFD）（n=5）。喂养8周后,观察肝脏光镜下改变,检测肝脏GSH、MDA、TNFα和IL-6水平。Western-Blot检测肝脏NF-kB蛋白表达水平,观察敲除Nrf2对肝脏NF-kB活性作用的影响。结果：1.光镜下观察,Control组与KO组小鼠肝脏结构无明显变化,HFD组小鼠肝脏呈现大片脂肪沉积和炎症细胞浸润,KOHFD组小鼠肝脏则呈现明显的大泡性变性,且炎症细胞浸润较HFD组明显加重;2.与Control组相比,KO组小鼠肝脏MDA轻度升高,GSH轻度降低,但无明显差异,而HFD组和KOHFD组小鼠肝脏MDA显著升高（P〈0.05）,GSH显著降低（P〈0.05）,且KOHFD组MDA明显高于HFD组（P〈0.05）,GSH明显低于HFD组（P〈0.05）。3.ELISA结果显示,与Control组相比,KO组小鼠肝脏TNFα和IL-6分泌轻度增加,而HFD组和KOHFD组小鼠肝脏TNFα与IL-6水平显著升高（P〈0.05）,且KOHFD组小鼠肝脏TNFα与IL-6显著高于HFD组（P〈0.05）;4.Western-Blot结果显示,Control组和KO组之间无明显差异,而KOHFD组和HFD组小鼠肝脏胞核NF-kB蛋白表达水平显著升高,且KOHFD组高于HFD组。结论：敲除Nrf2可以显著加重高脂饮食诱导的小鼠肝脏氧化应激水平,进而促进NF-kB的活化,从而为通过以Nrf2为靶点治疗NASH提供重要的实验依据。     

不同萃取体系对小球藻藻渣成分的影响

考察5种萃取体系（A：正己烷,B：正己烷/乙醇,C：正己烷/异丙醇,D：氯仿/甲醇,E：氯仿/乙醇）对小球藻（Chlorella phyrenoidosa）油脂的提取效果及藻渣成分的影响。实验结果表明：不同的萃取体系下,油脂得率为D（12.27%）、E（8.87%）、C（7.71%）、B（6.80%）、A（3.91%）,藻渣蛋白含量为A（52.60%）、E（46.23%）、B（40.19%）、C（39.52%）、D（32.52%）,藻渣碳水化合物含量为A（23.28%）、E（16.15%）、B（13.24%）、D（13.50%）、C（9.06%）;藻渣色素含量为A（1.75%）、E（1.29%）、B（1.14%）、C（0.96%）、D（0.58%）;藻渣灰分含量为D（3.63%）、E（2.94%）、C（2.23%）、B（2.25%）、A（1.48%）。综合考虑微藻生物柴油的生产及藻渣的可利用性,V（氯仿）/V（乙醇）=1是一种油脂萃取效果较好,藻渣营养成分损失较小的小球藻油脂萃取体系。     

Expression and epigenetic regulation of angiogenesis-related factors during dormancy and recurrent growth of ovarian carcinoma

The initiation of angiogenesis can mark the transition from tumor dormancy to active growth and recurrence. Mechanisms that regulate recurrence in human cancers are poorly understood, in part because of the absence of relevant models. The induction of ARHI (DIRAS3) induces dormancy and autophagy in human ovarian cancer xenografts but produces autophagic cell death in culture. The addition of VEGF to cultures maintains the viability of dormant autophagic cancer cells, thereby permitting active growth when ARHI is downregulated, which mimics the “recurrence” of growth in xenografts. Two inducible ovarian cancer cell lines, SKOv3-ARHI and Hey-ARHI, were used. The expression level of angiogenesis factors was evaluated by real-time PCR, immunohistochemistry, immunocytochemistry and western blot; their epigenetic regulation was measured by bisulfite sequencing and chromatin immunoprecipitation. Six of the 15 angiogenesis factors were upregulated in dormant cancer cells (tissue inhibitor of metalloproteinases-3, TIMP3; thrombospondin-1, TSP1; angiopoietin-1; angiopoietin-2; angiopoietin-4; E-cadherin, CDH1). We found that TIMP3 and CDH1 expression was regulated epigenetically and was related inversely to the DNA methylation of their promoters in cell cultures and in xenografts. Increased H3K9 acetylation was associated with higher TIMP3 expression in dormant SKOv3-ARHI cells, while decreased H3K27me3 resulted in the upregulation of TIMP3 in dormant Hey-ARHI cells. Elevated CDH1 expression during dormancy was associated with an increase in both H3K4me3 and H3K9Ac in two cell lines. CpG demethylating agents and/or histone deacetylase inhibitors inhibited the re-growth of dormant cancer cells, which was associated with the re-expression of anti-angiogenic genes. The expression of the anti-angiogenic genes TIMP3 and CDH1 is elevated during dormancy and is reduced during the transition to active growth by changes in DNA methylation and histone modification.     

Role of recombinant plasmid pEGFP-N1-IGF-1 transfection in alleviating osteoporosis in ovariectomized rats

Decreased levels of serum insulin-like growth factor-1 (IGF-1) have been proven to cause osteoporosis. Gene transfer of IGF-1 offers an attractive technology to treat skeletal metabolic disorders including osteoporosis, but the viral vectors are limited by their high antigenicity and immune response. Our purpose was to investigate the expression of a non-invasive vector, recombinant plasmid enhanced green fluorescent protein-N1 (pEGFP-N1) that transferred IGF-1 gene into ovariectomized (OVX) rats in vivo and evaluate the effect of this therapy on osteoporosis. OVX or sham operations were performed in 60 female, 7-month-old unmated SD rats. 12 weeks after OVX operation, the vectors were transfected to the 10-month-old rats and experimental data were detected from 48 h to 7 week after transfection. Our results showed that remarkable expression of fluorescence and serum IGF-1 was observed in the rats transfected by recombinant plasmids, indicating that IGF-1 gene was successfully transferred to OVX rats by injecting the vector through hydrodynamic method via the tail vein. The bone metabolism index including serum alkaline phosphatase, the histomorphometric parameters of lumbar vertebra including trabecular area percentage, trabecular thickness, trabecular number and trabecular separation, and the bone mineral density (BMD) and biomechanical parameters of lumbar vertebra including BMD, maximum condensing force, crushing strength in OVX rats transfected by pEGFP-N1-IGF-1 were improved remarkably compared with OVX+pEGFP-N1 rats, indicating that the transfection of recombinant plasmid pEGFP-N1-IGF-1 played a significant role in alleviating osteoporosis in rats induced by OVX. This encouraged a potential approach of IGF-1 gene therapy to the treatment of osteoporosis.