红肉脐橙(Citrus sinensis L.)果肉中特征色素提取方法探索

红肉脐橙是目前为止惟一果肉呈粉红色或红色的脐橙,其呈色色素尚未见报道。为深入研究其果肉中特征色素的构成及在果实生长发育和贮存过程中色素含量的变化,本研究在对该色素初步定性的基础上对提取方法进行了探索。在分析红肉脐橙果肉中色素萃取液的紫外可见吸收光谱时发现,该色素有类胡萝卜素的特征吸收峰,将萃取液进行薄层层析则进一步证实其内含有番茄红素。在色素提取的前处理方式上,本研究认为冷冻干燥样品的浸提效果较真空干燥的样品稳定(变异系数小),且浸提率相差无几,所以选择冷冻干燥作为合适的样品前处理方式。色素浸提条件的正交实验结果表明,最佳浸提条件为50℃、1.5h内浸提4次,浸提的料液比为1:60。此外,浸提温度、浸提时间、浸提料液比和浸提次数对浸提率都有显著影响,影响最大的因素是浸提温度,最佳温度为50℃,随温度降低浸提率显著降低。其次是侵提时间的长短,最佳浸提时间为1.5h,延长或缩短0.5h都会使浸提率下降。浸提次数对浸提率的影响是最小的。     

柑橘类果实汁胞的红色现象及其呈色色素

柑橘类果实中不乏汁胞为红色类型者,如红肉葡萄柚(Citrus paradisii Macf.)的粉红色是番茄红素所致,血橙(C.sinensis L.)的血红色来源于花青素,此外,还有红色的红江橙(C.sinensis L.)、橙红色的元红(C.reticulata Blan-co)和紫红色的五布红心柚(C.grandis L.)等,它们因色泽鲜艳而具备特殊的市场潜力,但呈色色素尚未见报道。在符合审美观的同时,红色汁胞品种还具有特殊的营养价值,如类黄酮、类胡萝卜素等色素富集可软化血管、抗癌、抗氧化及消除体内自由基等。综合有关文献,首次将国内外主要红色汁胞柑橘品种依宽皮柑橘、甜橙、柚、葡萄柚及杂柑等分类进行综述,分析可能的呈色色素,阐明导致红色性状产生的两类色素—花青素、类胡萝卜素生物合成途径     

In vitro growth inhibition effects of rhHGF/cHGF on SMMC-7721 human HCC cell line

AIM:To examine the effects of recombinant human hepatocyte growth factor(rhHGF) and native calf HGF(cHGF) on SMMC-7721 human hepatocellular carcinoma(HCC) cell line. METHODS:Human HCC cell line culture, photometric assay, and flow cytometric assay were used in this study .RESULTS:A similar type of dose-dependent cell growth inhibition effect on SMMC-7721 human HCC cells by rhHGF(5-20 μg/L) as well as by cHGF(25-100 mg/L) had been found, with the maximal effect at the highest concentration used. Approximately over 50% of the cells treated with rhHGF(5 μg/L, 10 μg/L, 20 μg/L) accumulated in the quiescent G₀/G₁ phase of the cell cycle over incubation periods for 3 d. CONCLUSION:The growth of SMMC-7721 human HCC cells was strongly inhibited by both rhHGF and cHGF. This might be because the cells exposed to HGF became arrested in the G₀/G₁ phase.     

Adult rat and human bone marrow mesenchymal stem cells differentiate into neurons with Musk's polypeptide

AIM: To investigate the differentiation from adult rat and human bone marrow mesenchymal stem cells (BMMSCs) into neuron with musk polypeptide (Mu-P).METHODS: Adult rat and human BMMSCs were induced with Mu-P.Neuron-specific enolase (NSE),neurofilament (NF),Nestin,glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry.RESULTS: Simple methods with Mu-P induced adult rat and human BMMSCs exhibiting a neuronal phenotype,expressing Nestin at 3 hours to 5 hours,and expressing NE and NF at 5 hours to 7 days.But the neuron-like cells didn't express the glial astrocyte marker GFAP.CONCLUSION: Adult rat and human BMMSCs can be induced to differentiate into neurons with Mu-P.     

Inhibitory effect of berberine on the activation and proliferation of T lymphocytes

AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 μmol/L and 50 μmol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 μmol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G₀/G₁ phase to S and G₂/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.     

Protein expression of cyclin D2 and p16 in proliferation and differentiation of cultured cardiac myocytes

AIM:To investigate the protein expression of cyclin D₂ and p16 in proliferation and differentiation of cultured cardiac myocytes.METHODS:One-day-old Sparague-Dawley rats were used. Cardiac myocytes(CM) were collected by a trypsin-dispersal method and cultured. Cell growth line and fluorescence activated cell sorting (FACS) were used to investigate the proliferation of CM. Ultra-thin sections were made to observe the ultrastructure of CM under transmission electron microscope. The expression of cyclin D₂ and p16 in CM were measured using immunocytochemistry and image analysis.RESULTS:①Results of cell growth line and FACS analysis showed that cultured CM could proliferate in the first 3 cultured days, but the ability decreased quickly, concomitant with differentiation. CM was obseved quiescent in cell cycle three days later. The ultrastructure of CM showed the large amount of myofilaments and mitochondrion. ②The protein expression of cyclin D₂ in 3,4,5 day CM group was 0.89 times(P＜0.05),0.80 times (P＜0.05) and 0.56 times (P＜0.01) of that in 1 day group, respectively. The expression of p16 in CM was increased during the culture process, 2,3,4,5 day group were 1.63 times, 1.72 times, 1.99 times and 2.84 times (P＜0.01) of that in 1 day group, respectively.CONCLUSION:Cultured neonatal rat cardiac myocytes could proliferate during the first 3 days after incubation, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D₂ and p16 play the key roles in CM postnatal development. Downregulation of cyclin D₂ and upregulation of p16 may induce CM differentiation.     

低温胁迫下弓葵幼苗膜脂过氧化及保护酶活性的变化

 低温胁迫下弓葵( Butia capitata Becc) 幼苗叶片的MDA 含量逐渐增加, 膜脂过氧化作用增强。- 8 ℃条件下的膜脂过氧化作用明显强于2 ℃。细胞膜透性在2 ℃条件下变化不大, - 8 ℃时则随低温胁迫时间延长而急剧上升, 细胞膜受到伤害。- 8 ℃胁迫下细胞保护酶SOD、POD 和CAT 活性短期(6 h) 内升高,然后下降, 24 h 以后3 种保护酶受到低温胁迫的严重抑制。在2 ℃胁迫下, SOD 活性在6 h 内变化不大, 随后下降; CAT活性变化趋势与- 8 ℃时相似, 但变化幅度较小; POD 虽也呈现先升后降的趋势, 但降幅明显小于升幅, 至48 h 时POD 活性仍维持较高水平。2 ℃低温胁迫不是抑制而是促进POD 活性的提高。     

红阳猕猴桃全红芽变系的RAPD分析

 以10 bp 的随机引物, 通过PCR 技术对猕猴桃全红芽变系86-3 及对照品种红阳的基因组DNA进行扩增, 以探讨芽变系遗传物质的变化。以86-3 母株与红阳为材料对300 条单引物进行了筛选, 得到在两个样品间的扩增有多态性片段10 条引物( S107、S84、S157、S27、S188、AY17、AY5、R11、AV9、AM11) 。10 条引物共扩增出148 个片段, 多态性片段为20 个, 占13.5 %。两样品间相似系数为0.7619 , 遗传距离为0.1429 , 表明86-3 果肉颜色的变化与遗传物质的改变密切相关。用4 种引物S107、S84、S157、AY17 分别对芽变系单株及对照进行RAPD 扩增, 不同树龄、不同砧木的芽变单株与对照间的特异性片段同引物筛选中所得结果一致, 可用所得引物对全红芽变系嫁接株的果肉颜色进行早期预测。     

肥城桃两品系果实细胞壁成分和水解酶活性的比较

 随着果实发育成熟，肥城桃两品系果实中细胞壁含量不断下降。可溶性果胶从8月1～16日明显增加，这时果肉硬度下降最快。两品系果实的细胞壁中离子结合果胶和纤维素含量下降比例不同，这可能是造成两品系成熟软化特性不同的主要因素之一。果实发育后期‘白里肥城桃’和‘红里肥城桃’纤维素酶活性变化的不同，意味着纤维素酶在肥城桃果实软化中起重要作用。肥城桃果实中没有检测到内切PG活性，成熟后期外切PG活性上升较快。