NFT系统和松树皮基质栽培对草莓干物质积累的影响

在温室采用 NFT系统和松树皮基质栽培法培植草莓 (seascape和 pajaro)。分别测定其叶片数、鲜重和干重 ,结果数、鲜重和干重 ,分蘖数、鲜重和干重 ,根长和干重及其地上部分与地下部分的比值。在 NFT系统和松树皮基质中 ,seascape草莓营养器官的干物质积累差异明显 ;被清除果实后 ,其叶面积的生长和新的坐果率成负相关 ,叶面积的同期增长速率较大 ;seascape和 pajaro草莓生殖器官的干物质积累无差异 ;pajaro草莓的地上和地下部分干物质积累无明显差异。在 NFT系统中 ,不同时间定植的 pajaro,其干物质积累差异不显著 ,但定植时间晚影响其坐果 ;seascape干物质积累量小于树皮基质中的积累量。 NFT和松树皮基质栽培方式对 seascape和 pajaro草莓根的影响较大。

脱细胞鱼皮基质作为新型组织工程支架的研究进展

目的分析脱细胞鱼皮基质作为新型组织工程支架材料的最新研究进展,讨论该材料用于医学临床的可行性及风险管理。方法基于对脱细胞鱼皮基质研究最新进展的广泛调研,综合分析其研发、应用现况,提出应重点关注的关键问题。结果脱细胞鱼皮基质作为新型细胞外基质有巨大的临床应用潜力,但我国在该领域的研究几乎空白,国外对该类产品的转化医学研究刚刚兴起,主要集中于人工皮肤、外科补片、创伤敷料等方面。结论脱细胞鱼皮基质作为组织工程支架新产品的开发具有良好的临床可行性和必要性,但作为新策略、新产品尚需进行大量的应用基础研究。

脱细胞鱼皮基质的制备、改性与应用前景

脱细胞基质(ACM)是指不同来源的组织或器官经过脱细胞处理后,去除细胞等抗原性较强的组分,保留由胶原蛋白组成的纤维网状结构。因为其较低的免疫原性与较高的生物相容性,常被用于再生医学和组织修复中。脱细胞鱼皮基质作为一种新型生物支架材料,与传统的哺乳动物脱细胞真皮基质相比,具有治疗效果好且不容易会引起炎症反应等优点。因此,主要阐述脱细胞鱼皮基质的制备、改性及其应用前景,并对其制备过程中存在问题进行讨论。

Isolation, Cuture and Biological Characteristic Analysis of Stromal and Glandular Epithelial Cells of Buffalo (Bubalus bubalis) Endometrium

[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.

Possible involvement of integrin signaling pathway in the process of recovery from restraint stress in rats

Objective To search novel genes or pathways involved in the recovery process after restraint stress in rats. Methods We compared the hypothalamus transcriptional profiles of two different recovery patterns （fast recovery vs slow recovery） from restraint stress in rats using oligonucleotide microarray, the recovery pattern was determined by the decrement of plasma adrenocorticotropic-hormone （ACTH） and corticosterone levels during one hour recovery period after stress. A real-time quantitative RT-PCR was applied to validate the differential expressed genes. Results Analysis of the microarray data showed that most of genes were not differentially expressed between fast recovery group and slow recovery group. Among the differentially expressed genes we found that talin, together with serine/threonine protein phosphatase PPl-beta catalytic subunit （PP-1B） and integrin α-6 precursor （VLA-6） genes, were at least 1.5 fold upregulated in the fast recovery group, while junctional adhesion molecule 1 （F11r） was 1.5 fold down-regulated in the fast recovery group. Conclusion The results implied that integrin signaling pathway may be involved in the recovery from restraint stress in rats. The present study provided a global overview of hypothalamus transcriptional profiles during the process of recovery from the restraint stress in rats. The integrin signaling pathway seems to be involved in the recovery process, which deserves further study to clarify the integrin-mediated recovery mechanism after restraint stress.

Study on Multiplication, Rooting and Transplanting of Tissue Culture Plantlets of Rhododendron chrysanthum Pall

[Objective] This study aimed to shorten the multiplication culture and root- ing culture periods of Rh. chrysanthum Pall. [Method] The Rh. chrysanthum Pall tis- sue culture plantlets collected from Changbai Mountain were used as material, and the effects of different hormone combinations and coconut milk on the proliferation and differentiation of Rh. chrysanthum Pall tissue culture plantlets were investigated. In addition, the rooting medium and transplanting matrix for Rh. chrysanthum Pall tissue culture plantlets were explored. [Result] The medium composed of modified MS, iBA （3 mg/L） and ZT （1.5 mg/L） was the optimum medium for subculture mul- tiplication of Rh. chrysanthum Pall tissue culture plantlets. The multiplication multiple and average plant height were significantly improved by adding coconut milk into the medium （150 mg/L）. [Conclusion] For Rh. chrysanthum Pall tissue culture plantlets, the optimum rooting culture medium was composed of modified MS （1/4） and IBA （5.0 mg/L）, and the tissue culture plantlets began to root 8 d after the inoculation. The root induction treatment was carried out after a 15-d sand culture, and the suitable matrix was composed of tufty soil, humus soil and perlite （2：1：1） with a survival rate of 95.66%.

Proteomic analysis of pancreatic intraepithelial neoplasia and pancreatic carcinoma in rat models

AIM:To detect the proteomic variabilities of pancreatic intraepithelial neoplasia(PanIN)and pancreatic carcinoma(PC)induced by 7,12-dimethylbenzanthracene(DMBA) in rat models and to identify potential biomarkers.METHODS:Sixty adult male Sprague Dawley rats were randomized into three groups.The rats had DMBA implanted into their pancreas for one(n=20)or two months(n=20)or assigned to the normal group(n =20).The rats were killed after one or two months,and were evaluated histopathologically.Three tissue samples from each group of rats with either normal pancreas,PanIN(PanIN-2)or PC were examined by 2D-DIGE.The different expression spot features were analyzed by matrix-assisted laser desorption/ionizationtime of flight/time of flight(MALDI-TOF/TOF)tandem mass spectrometry.The expression of enolase 1,a differentially expressed protein,was identified by immu-nohistochemistry.RESULTS:There was significant difference in the proportions of neoplastic changes between the 1-and 2-mogroups(P=0.0488).There was an increase in the frequency of adenocarcinomas in the 2-mo group compared with the 1-mo group(P=0.0309).No neoplastic changes were observed in any of the animals in the normal group.Enolase 1,pancreatic ELA3B,necdin,Hbp23,CHD3,hnRNP A2/B1,Rap80,and Gnb2l1 were up-regulated in the PanIN and PC tissues,and CEL,TPT1,NME2,PCK2,an unnamed protein product,and glycine C-acetyltransferase were down-regulated in the PanIN and PC tissues.The immunohistochemical results showed that enolase 1 expression was up-regulated in the pancreatic cancer tissues of rats and humans.CONCLUSION:The pancreatic protein expression changes induced by DMBA suggest potential molecular targets for the early diagnosis and treatment of PC.

Targeting of RhoE inhibits epithelial-mesenchymal transition during colorectal cancer cell migration

Objective Despite microRNA （miR-200b） being proved to promote the proliferation of colorectal cancer （CRC） cells, the relationship between miR-200b and epithelial-mesenchymal transition （EMT） of CRC cells remains poorly understood. The aim of the study was to investigate the relationship between miR-200b and EMT during CRC cell migration. Methods The effect of miR-200b on EMT-associated markers E-cadherin and vimentin was evaluated by western blot in CRC cells （SW620 and HT-29） by treatment with miR-200b mimics and inhibitors. A lucifer- ase reporter assay was employed to detect downstream targets of miR-200b. Transwell migration assays were used to detect CRC cell migration. Results Westem blots revealed that treatment with miR-200b mimics led to up-regulation of E-cadherin and down-regulation of vimentin, metalloproteinase （MMP）-9, and MMP-2, whereas treatment with miR- 200b inhibitor exhibited opposite effects on expression of E-cadherin and vimentin. Luciferase reporter assays demonstrated that RhoE （RND3） was targeted by miR-200b. Two predicted target sites of miR-200b were present in the 3＇-UTR of RhoE. Predicted target site 1 was from nucleotides 1584 to 1591, and site 2 was from nucleotides 1729 to 1735. RhoE knockdown cell lines were also established to investigate the impact of RhoE and miR-200b on EMT and cell migration. RhoE knockdown enhanced the effect of miR- 200b mimics, up-regulating E-cadherin and down-regulating vimentin. RhoE knockdown also inhibited cell migration. Furthermore, miR-200b mimic treatment further promoted the inhibitory effect of RhoE knock- down on cell migration.

Epithelial-mesenchymal transition and cancermetastasis

Epithelial-mesenchymal transition （EMT） is initially considered as a physiological phenomenon during the embryogenesis of mammals, as well as a basic biological event maintaining the stability of the vital body. Recent researches indicated that EMT plays a critical role in various tumors progression, through which epithelial cancers invade and metastasize. The cell characteristics are changed during EMT, in which the cells lose cell-cell and cell-matrix interactions and apical polarity, reorganize their cytoskeleton, and become isolated, motile, as well as resistant to anoikis, then become spindle-shaped mesenchymal cells. This review lays emphasis on studying the cell morphogenesis, makers and molecular mechanism regulation about EMT, discussing the relationship between the EMT and the cancer development and metastasis.

Melanocortin-1 receptor gene variants in four Chinese ethnic populations

There is strong relationship between melanocortin-1 receptor (MCIR) gene variants and human hair color and skin type. Based on a sequencing study of MC1R gene in 50 individuals from the Uygur, Tibetan, Wa and Dai ethnic populations, we discuss the occurrence of 7 mc1r variants consisting of 5 nonsynonymous sites (Val60Leu, Arg67Gln, Val92Met, Arg163Gln and Ala299Val) and 2 synonymous sites (C414T and A942G), among which C414T and Ala299Val were reported for the first time. Confirmation and analysis were the made of 122 individuals at three common point mutations (Val92Met, Arg163Gln, A942G) using PCR-SSCP. The frequency of Arg163Gln variant varies in the four ethnic populations, with percentage of 40%, 85.0%, 66. 2% and 72.7%, respectively, while those of Val92Met and A942G are roughly similar in these four populations. The different environments, migration and admixture of various ethnic groups in China might have impact on the observed frequency of Arg163Gln.

Significance of glucocorticoid receptor expression in colonic mucosal cells of patients with ulcerative colitis

AIM: Glucocorticoid (GC) resistant ulcerative colitis (UC) remains a serious disease and is difficult to manage. Although the molecular basis of GC insensitivity is still unknown, GC receptors (GRAAA and GRp) may play an important role in it. This study was aimed to investigate the relationship between the expression of GRa and GRp in colonic mucosal cells of patients with UC, the efficacy of GC therapy and the intensity of inflammation. METHODS: Twenty-five cases of UC were classified into: GC sensitive (n = 16) and GC resistant (n - 9) cases. Patients consisted of mild (n = 6), moderate (n = 8) and severe (n = 11) cases. GRa and GRp expression in colonic mucosal specimens were investigated by immunohistochemistry, and compared between GC resistant and sensitive groups, and also among various degrees of inflammation. RESULTS: All cases were positive for GRa and GRp expression. Both positive association between GRa expression and the response of UC to GC and strong negative association between GRp expression and the response of UC to GC were identified. There was no significant association between GRa/GRp expression and the degree of inflammation of UC. CONCLUSION: These findings suggest that both GRa and GRp may play an important role in the action of GC, and that GRp functions as a dominant negative inhibitor of GRa. Expression of GRa and GRp in colonic mucosal cells of patients with UC may serve as predictors of glucocorticoid response, but can not function as markers of inflammatory intensity.

Esophageal tissue engineering:A new approach for esophageal replacement

A number of congenital and acquired disorders require esophageal tissue replacement.Various surgical techniques,such as gastric and colonic interposition,are standards of treatment,but frequently complicated by stenosis and other problems.Regenerative medicine approaches facilitate the use of biological constructs to replace or regenerate normal tissue function.We review the literature of esophageal tissue engineering,discuss its implications,compare the methodologies that have been employed and suggest possible directions for the future.Medline,Embase,the Cochrane Library,National Research Register and ClinicalTrials.gov databases were searched with the following search terms:stem cell and esophagus,esophageal replacement,esophageal tissue engineering,esophageal substitution.Reference lists of papers identified were also examined and experts in this field contacted for further information.All full-text articles in English of all potentially relevant abstracts were reviewed.Tissue engineering has involved acellular scaffolds that were either transplanted with the aim of being repopulated by host cells or seeded prior to transplantation.When acellular scaffolds were used to replace patch and short tubular defects they allowed epithelial and partial muscular migration whereas when employed for long tubular defects the results were poor leading to an increased rate of stenosis and mortality.Stenting has been shown as an effective means to reduce stenotic changes and promote cell migration,whilst omental wrapping to induce vascularization of the construct has an uncertain benefit.Decellularized matrices have been recently suggested as the optimal choice for scaffolds,but smart polymers that will incorporate signalling to promote cell-scaffold interaction may provide a more reproducible and available solution.Results in animal models that have used seeded scaffolds strongly suggest that seeding of both muscle and epithelial cells on scaffolds prior to implantation is a prerequisite for complete esophageal replacement.Novel approaches need to be designed to allow for peristalsis and vascularization in the engineered esophagus.Although esophageal tissue engineering potentially offers a real alternative to conventional treatments for severe esophageal disease,important barriers remain that need to be addressed.

Preliminary study of therapeutic effects of panax notoginside and methylprednisolone on pulmonary fibrosis in rats

Objective: To study the therapeutic effects of panax notoginside (PN) and methylprednisolone (MP) on pulmonary fibrosis in rats. Methods: Bleomycin was introduced into the bronchial tree of 75 Wistar male rats through a tracheal incision to establish a rat model of pulmonary fibrosis (PF). The rats were equally divided into 3 groups: PF group, PN treated group and MP treated group. Five rats of each group were killed 1, 3, 7, 14 and 28 d after the administration of bleomycin and the specimens of lung tissue and plasma were collected for the determination of content of collagen Ⅰ and Ⅲ with immunohistochemical method and the level of MIP-1α and MCP-1 with ELIZA. Results: The severity of PF, the content of collagen Ⅰ and Ⅲ and the level of MIP-1α and MCP-1 were significantly decreased in PN and MP treated groups than in PF group (P<0.05). Conclusion: PN and MP are effective to control the development of PF induced with bleomycin in rats.

Effect of quercetin on expression of matrix metallo - proteinases and tissue inhibi tor of matalloproteinase -1 in cultured rat hepatic stellate cells

Objective: To study the effects of quercetin (QU) on matrix metallo-proteinases (MMPs) , the tissue inhibitor of matalloproteinase-1 (TIMP-1) , procollagen I and 2 proteoglycans (decorin and biglycan) mRNA expression in cultured rat hepatic stellate cell line HSC-T6 cells. Methods: Cells were treated with different concentrations of QU (12. 5, 25, 50 μmol/L) or drug solvent (0. 1 % Me2SO) for 24 h. mRNA expression was determined by reverse transcription polymerase chain reaction (RT-PCR). Results: QU (12.5 - 50 μmol/L) enhanced collagenase (rat MMP-13) and membrane typel-MMP (MMP-14) mRNA expression, decreased procollagen I mRNA expression in a concentration-dependent manner, but did not affect gelatinase-A (MMP-2) , TIMP-1, decorin and biglycan expression. Conclusion: QU may decrease matrix deposition and increase matrix degradation, which might be beneficial to liver fibrosis.

Vascular endothelial growth factor 165b expression in stromal cells and colorectal cancer

AIM:To characterize the implications of vascular endothelial growth factor(VEGF)-A in stromal cells and colorectal cancer and the expression of VEGF-A splice variants.METHODS:VEGF-A expression in tumor and stromal cells from 165 consecutive patients with colorectal cancer was examined by immunohistochemistry.The association between VEGF-A expression status and clinicopathological factors was investigated.Twenty freshfrozen samples were obtained for laser capture microdissection to analyze the splice variants of VEGF-A.RESULTS:VEGF-A was expressed in 53.9% and 42.4% of tumor and stromal cells,respectively.VEGF-A expression in tumor cells(t-VEGF-A) was associated with advanced clinical stage(stage 0,1/9;stage 1,2/16;stage 2,32/55;stage 3,38/66;stage 4,16/19,P < 0.0001).VEGF-A expression in stromal cells(s-VEGF-A) increased in the earlier clinical stage(stage 0,7/9;stage 1,6/16;stage 2,33/55;stage 3,22/66;stage 4,5/19;P = 0.004).Multivariate analyses for risk factors of recurrence showed that only s-VEGF-A expression was an independent risk factor for recurrence(relative risk 0.309,95% confidence interval 0.141-0.676,P = 0.0033).The five-year disease-free survival(DFS) rates of t-VEGF-A-positive and-negative cases were 51.4% and 62.9%,respectively.There was no significant difference in t-VEGF-A expression status.The five-year DFS rates of s-VEGF-A-positive and-negative cases were 73.8% and 39.9%,respectively.s-VEGFA-positive cases had significantly better survival than s-VEGF-A-negative cases(P = 0.0005).Splice variant analysis revealed that t-VEGF-A was mainly composed of VEGF165 and that s-VEGF-A included both VEGF165 and VEGF165b.In cases with no venous invasion(v0),the level of VEGF165b mRNA was significantly higher(v0 204.5 ± 122.7,v1 32.5 ± 36.7,v2 2.1 ± 1.7,P = 0.03).The microvessel density tended to be lower in cases with higher VEGF165b mRNA levels.CONCLUSION:s-VEGF-A appears be a good prognostic factor for colorectal cancer and includes VEGF165 and VEGF165b.

The effect of alphastatin peptide suppressing the hypoxia-induced vasculogenic mimicry formation of glioma

Objective:To investigate the vasculogenic mimicry formation induced by hypoxia in Ⅱ-Ⅲ human glioma cell and the effect of alphastatin peptide suppressing the hypoxia-induced vasculogenic mimicry formation and the mechanism.Methods:MTT,Transwell and three-dimentional culture were used to detect the proliferation,migration and tubule formation of SHG44.The expression of vascular endothelial growth factor-α(VEGF-α),erythropoietin-producing hepatocellular carcinoma-A2 (EphA2) and matrix metalloproteinases 2 (MMP2) was detected by RT-PCR and Western blotting analysis.Results:The OD 490 in hypoxia group was 0.60±0.06 and in control group was 0.46±0.05.The number of cell migration was 178.71±18.81 in hypoxia group and 85.86±17.92 in control group.The tubule formation was 56.80±12.21 in hypoxia group and 4.20±2.62 in control group.The proliferation,migration and tubule formation in hypoxia group were significantly higher than that in control group.The expression of VEGF-α,EphA2 and MMP2 was upregulated in hypoxia.When various concentrations of alphastatin (100,1 000,10 000 nmol/L) were added to hypoxia group,the numbers of cell migration were 142.57±12.12,92.71±17.68,30.00±7.72 and the tubule formation were 47.71±10.58,18.86±8.40,8.43±5.62.The cell migration and tubule formation were significantly suppressed by alphastatin in a dose-dependent manner.In alphastatin group,the phosphorylation of EphA2 protein (P=0.037,F=4.629) and activation of MMP2 protein (P=0.005,F=9.331) were significantly suppressed but there was no change in VEGF-α protein.Conclusion:Ⅱ-Ⅲ human glioma cell is able to form vasculogenic mimicry induced by hypoxia and alphastatin peptide can suppress the hypoxia-induced vasculogenic mimicry.VEGF-α induced EphA2 phospharilation and MMP2 activation maybe the key pathway to form vasculogenic mimicry.

New record of Lobophora rosacea(Dictyotales; Phaeophyceae) from the South China Sea

Lobophora rosacea C.W. Vieira, Payri et De Clerck is reported from the South China Sea for the first time. Our specimens are very similar to L. rosacea recently described from New Caledonia, not only in morphology but also in rbcL and cox3 gene sequences. The fan-shaped thallus grows erectly, attaching to the substrate by a basal holdfast. The thallus is composed of a single layer of large medullary cells and three to four layers of cortical cells on both sides of the medulla. Mature sporophytes are detected, with sporangium sori scattered on both surfaces of the thallus.

In vitro effect of p2l^(WAF-1/CIP1) gene on growth of human glioma cells mediated by EGFR targeted non-viral vector GE7 system

Objective: To construct the EGFR targeted non-viral vector GE7 system and explore the in vitro effect of p21WAF-1/CIPI gene on growth of human glioma cells mediated by the GE7 system. Methods: The EGFR targeted non-viral vector GE7 gene delivery system was constructed. The malignant human glioma cell line U251MG was transfected in vitro with β-galactosidase gene ( reporter gene) and p21WAF-1/CIPI gee (therapeutic gene) using the GE7 system. By means of X-gal staining, MTS and FACS, the transfection efficiency of exogenous gene and apoptosis rate of tumor cells were examined. The expression of p21WAF-1/ CIPI gene in transfected U251MG cell was examined by immunohistochemis-try staining. Results: The highest transfer rate of exogenous gene was 70% . After transfection with p21WAF-1/CIPI gene, the expression of WAF-1 increased remarkably and steadily; the growth of U251MG cells were inhibited evidently. FACS examination showed G1 arrest. The average apoptosis rate was 25.2%. Conclusion: GE7 system has the ability to transfer exogenous gene to targeted cells efficiently, and expression of p21WAF-1/CIPI gene can induce apoptosis of glioma cell and inhibit its growth.

Application of modified polyethylene glycol hydrogels in the construction of tissue engineered heart valve

To enhance the adhesion of seeding-cells to the biomaterial scaffolds, the PEG-hydrogels were modified. Porcine aortic valves were decellularized with Triton X-100 and trypsin. The cells were encapsulated into the PEG-hydrogels to complete the process of the cells attaching to the acellular porcine aortic valves. Herein, the autologous mesenchymal stem cells （MSCs） of goats were selected as the seeding-cells and the tendency of MSCs toward differentiation was observed when the single semilunar TEHV had been implanted into their abdominal aortas. Furthermore, VEGF, TGF-β1, and the cell adhesive peptide motif RGD were incorporated. Light and electron microscopy observations were performed. Analysis of modified PEG-hydrogels TEHV＇s （PEG-TEHV） tensile strength, and the ratio of reendothelial and mural thrombosis revealed much better improvement than the naked acellular porcine aortic valve （NAPAV）. The data illustrated the critical importance of MSC differentiation into endothelial and myofibroblast for remodeling into native tissue. Our results indicate that it is feasible to reconstruct TEHV efficiently by combining modified PEG-hydrogels with acellular biomaterial scaffold andautologous MSCs cells.

Theoretical basis of a beneficial role for vitamin D in viral hepatitis

Abnormal bone metabolism and dysfunction of the calcium-parathyroid hormone-vitamin D axis have been reported in patients with viral hepatitis. Some studies suggested a relationship between vitamin D and viral hepatitis. Genetic studies have provided an opportunity to identify the proteins that link vitamin D to the pathology of viral hepatitis (i.e., the major histocompatibility complex class Ⅱ molecules, the vitamin D receptor, cytochrome P 450 , the renin-angiotensin system, apolipoprotein E, liver X receptor, toll-like receptor, and the proteins regulated by the Sp1 promoter gene). Vitamin D also exerts its effects on viral hepatitis via non-genomic factors, i.e., matrix metalloproteinase, endothelial vascular growth factor, prostaglandins, cyclooxygenase-2, and oxidative stress. In conclusion, vitamin D could have a beneficial role in viral hepatitis. Calcitriol is best used for viral hepatitis because it is the active form of the vitamin D 3 metabolite.