光合菌HP1胞外多糖对乌鳢肌肉营养成分的影响

唐家毅; 段元慧; 汪小军; 钱雪桥

doi:10.19303/j.issn.1008-0384.2019.05.009

光合菌HP1胞外多糖对乌鳢肌肉营养成分的影响

doi: 10.19303/j.issn.1008-0384.2019.05.009

唐家毅^{1, 2,},
段元慧¹,
汪小军²,
钱雪桥^1, ,

1.
广东海大集团股份有限公司畜牧水产研究中心/农业部微生态资源养殖利用重点实验室, 广东广州 511400
2.
华南理工大学环境与能源学院, 广东广州 510006

基金项目:

广东省应用型科技研发及重大科技成果转化专项 2017B020236004

详细信息

作者简介:
唐家毅(1982-), 男, 博士, 研究方向:微生物应用技术(E-mail:jiaqian2001@163.com)

通讯作者:
钱雪桥(1967-), 男, 博士, 教授级高工, 研究方向:水生动物营养(E-mail:qxq@haid.com.cn)

中图分类号: S949
计量
- 文章访问数: 1074
- HTML全文浏览量: 171
- PDF下载量: 23
- 被引次数: 0
出版历程
- 收稿日期: 2019-02-27
- 修回日期: 2019-04-17
- 刊出日期: 2019-05-28

Nutritional Enhancement of Polysaccharides from Photosynthetic Bacteria HP1 on Ophiocephalusargus Meat

1.
Animal Husbandry and Fisheries Research Center of Guangdong Haid Group Co., Limited, Key Laboratory of microecological resources and utilization in breeding industry, Ministry of Agriculture Guangzhou, Guangdong 511400, China
2.
College of Environment and Energy, South China University of Technology, Guangzhou, Guangdong 510006, China

摘要

摘要: 目的筛选产多糖光合菌，为开发廉价易制备的益生菌多糖及应用提供参考。方法从虾塘中筛选一株产多糖的光合菌HP1，进行生理生化鉴定和16S rDNA序列比对。通过醇沉的方式从发酵液中制备光合菌HP1粗多糖，拌喂乌鳢30 d，以黄芪多糖作为参照，分别对比各个试验组对乌鳢肌肉营养成分的影响。结果 HP1为粪红假单胞菌Rhodopseudomonasfaecalis。HP1胞外多糖组对乌鳢肌肉pH值没有显著影响（P>0.05），但能显著降低肌肉失水率并促使肌纤维直径变细（P < 0.05）。拌喂HP1胞外多糖对乌鳢粗蛋白含量无显著影响（P>0.05），但对乌鳢肌肉中的氨基酸组成有显著影响（P < 0.05），分别提高乌鳢肌肉中的总氨基酸、必需氨基酸、呈味氨基酸含量8.7%、11%、9%。另外，拌喂HP1胞外多糖也分别提高了乌鳢肌肉的粗脂肪含量、二十碳五烯酸（EPA）和二十二碳六烯酸（DHA）总含量10.7%和12.8%，显著提高肌肉中n-3与n-6系列脂肪酸比值。结论拌喂HP1胞外多糖不仅提高乌鳢肌肉的食用口感，也提升了乌鳢肌肉的营养价值，其效果与黄芪多糖拌喂的效果相似。
- 乌鳢 /
- 粪红假单胞菌 /
- 胞外多糖 /
- 肌肉营养
Abstract: Objective Enhancement on the development and application of exopolysaccharides produced economically from photosynthetic bacteria fermentation was investigated. Method A strain of polysaccharide-producing photosynthetic bacteria, HP1, was isolated from a shrimp aquaculture pond. Crude polysaccharides were prepared from fermentation broth and precipitated by ethanol. Using astragalus polysaccharides for comparison, effects of inclusion of the polysaccharides in feed on nutrients in the meat of Ophiocephalus argus fed for 30 d were examined. Result The physiological and biochemical characteristics as well as 16S rDNA sequencing of HP1 identified it to be Rhodopseudomonas faecalis. The polysaccharides produced from HP1 fermentation had no significant effect on muscle pH (P>0.05), but significantly decreased the drip loss and muscle fiber diameter of the fish after the feeding (P < 0.05). In the meat, the crude protein did not differ significantly by the diet change (P>0.05), but the amino acids did (P < 0.05). Compared with control without polysaccharide addition in the forage, the contents of total, essential, and delicious amino acids in the fish muscles increased by 8.7%, 11%, and 9%, respectively, crude fats by 10.7%, EPA+DHA by 12.8%, and the ratio of n-3/n-6. Conclusion It appeared that a diet included the exopolysaccharides not only improved the eating quality but also enhanced the nutritional value of O. argus with effects comparable to astragalus polysaccharides.
- Ophiocephalus argus /
- Rhodopseudomonas faecalis /
- exopolysaccharide /
- muscle nutrient content

HTML全文

图 1 菌株HP1

注：A为菌落形态，B为菌体形态。

Figure 1. Morphology of HP1

Note:A.Colonial morphology, B.Cell.

下载: 全尺寸图片幻灯片

表 1 乌鳢肌肉感官评价参考标准

Table 1. Reference standards for sensory evaluation on O. argus meat

项目 Items	最好9~10分 Best (9-10 points)	较好6~8分 Better (6-8 points)	一般3~5分 Ordinary (3-5 points)	最差0~2分 Worst (0-2 points)
弹性Elasticity	肉质弹性好	肉质有弹性	肉质微有弹性	肉质杂乱无弹性
嫩度Tenderness	咀嚼时鲜嫩无残渣	咀嚼时鲜嫩，微有残渣	咀嚼时有粗糙感，有少量残渣	咀嚼时有粗糙感，有较多残渣
鲜味Fresh	鱼香味浓郁，鲜美	鱼香味较浓郁，较鲜	鱼香味较淡，微有鲜味	鱼香味极淡，无鲜味
甜味Sweet	甜味明显	微有甜味	甜味极淡	无甜味

下载: 导出CSV

表 2 菌株HP1生理生化特征

Table 2. Physiological and biochemical characteristics of HP1

项目 Items	结果 Results
最适温度 Optimum temperature	34~37℃
最适pH值 Optimum pH	6.5~8.5
2%耐盐试验 2% Salt tolerance test	+
5%耐盐试验 5%Salt tolerance test	+
7%耐盐试验 7% Salt tolerance test	―
明胶液化试验 Gelatin liquefaction test	―
甲基红试验 Methyl red test	―
过氧化氢酶试验 Catalase test	+
甘露醇 Mannitol	+
酪素 Casein	―
淀粉 Starch	―
柠檬酸盐 Citrate	―
葡萄糖 Glucose	+
乙酸钠 Sodium acetate	+
丁酸钠 Sodium butyrate	+
丙酸盐 Propionate	+

下载: 导出CSV

表 3 菌株HP1的16S rDNA序列在GenBank的比对结果

Table 3. 16S rDNA sequence alignment of HP1 in comparison to GenBank database

编号 Serial numbers	菌株 Strain	序列覆盖率 Sequence coverage/%	相似性 Similarity /%
NR_024971.1	Rhodopseudomonas faecalis	100	99.44
HQ912770.1	Rhodopseudomonas sp.	100	99.09
HQ154127.1	Rhodopseudomonas faecalis	100	99.02
EU410078.1	Rhodopseudomonas faecalis	100	98.95

下载: 导出CSV

表 4 拌喂HP1胞外多糖后乌鳢肉质感官评价

Table 4. Sensory evaluation on meat of O. argus fed with HP1 exopolysaccharides

项目 Items	C1	C2	C3
弹性Elasticity	5.0±0.2b	5.3±0.1b	6.2±0.3a
嫩度Tenderness	4.8±0.1b	6.3±0.5a	6.6±0.2a
鲜味Fresh	4.3±0.2b	5.7±0.1a	5.8±0.1a
甜味Sweet	4.2±0.3b	5.5±0.1a	5.9±0.3a
注：每组中同行平均值后不同字母表示差异显著(P＜0.05)。表 5~8同。 Note：Data with different lowercase letters on the same line indicate significant difference at P＜0.05. The same as Table 5-8.

下载: 导出CSV

表 5 HP1胞外多糖对乌鳢肌肉理化性质影响

Table 5. Effect of HP1 exopolysaccharides on physicochemical properties of O. argus meat

项目 Items	C1	C2	C3
肌纤维直径Diameter of muscle fiber/μm	75.3±2.1a	53.8±1.2b	56.7±2.8b
肌肉pH值pH of muscle	4.71±0.09	4.67±0.07	4.74±0.12
肌肉失水率Water loss rate of muscle /%	22.8±1.5a	18.4±1.1b	18.3±0.9b

下载: 导出CSV

表 6 HP1胞外多糖对乌鳢肌肉成分的影响

Table 6. Effect of HP1 exopolysaccharides on nutrient composition of O. argus muscles

项目 Items	C1	C2	C3
肌肉粗脂肪Crude fat of muscle /%	7.52±0.35b	7.74±0.19b	8.48±0.53a
肌肉粗蛋白质Crude protein of muscle /%	88.69±6.83	88.20±5.27	87.12±5.15
单尾末重Weight of single one/g	647±23	651±18	655±19

下载: 导出CSV

表 7 HP1胞外多糖后对乌鳢肌肉氨基酸组成的影响

Table 7. Effect of HP1 exopolysaccharides on amino acids in O. argus muscles

(单位/%)
种类 Species	氨基酸名称 Names of amino acid	C1	C2	C3
甜味氨基酸Sweet amino acid	苏氨酸Thr	3.56±0.08b	3.92±0.03a	3.94±0.06a
	脯氨酸Pro	3.49±0.04	3.51±0.11	3.60±0.18
	赖氨酸Lys	7.19±0.07b	7.99±0.13a	7.93±0.15a
	丝氨酸Ser	3.64±0.15	3.70±0.10	3.70±0.07
	甘氨酸Gly	3.88±0.12b	4.30±0.09a	4.27±0.05a
	丙氨酸Ala	4.38±0.05b	4.89±0.17a	4.87±0.12a
鲜味氨基酸Fresh amino acid	天冬氨酸Asp	8.13±0.12b	8.75±0.19a	8.88±0.24a
	谷氨酸Glu	10.42±0.26b	11.69±0.16a	11.53±0.12a
含硫氨基酸Sulfur containing amino acid	胱氨酸Cys	0.94±0.15	1.03±0.18	1.08±0.10
	蛋氨酸Met	2.07±0.18b	2.39±0.17a	2.38±0.07a
其他氨基酸Other amino acid	组氨酸His	2.25±0.04	2.28±0.12	2.38±0.08
	缬氨酸Val	3.82±0.05b	4.17±0.09a	4.13±0.06a
	异亮氨酸Iso	3.68±0.12b	4.03±0.05a	4.00±0.12a
	亮氨酸Leu	6.13±0.17b	6.88±0.12a	6.85±0.14a
	酪氨酸Tyr	2.45±0.05	2.52±0.09	2.48±0.06
	苯丙氨酸Phe	3.45±0.13b	3.92±0.04a	3.93±0.08a
	精氨酸Arg	4.78±0.05	4.83±0.21	4.84±0.16
	氨基酸总和∑TAA	74.26±0.11b	80.8±0.12a	80.79±0.11a
	人体必需氨基酸总和∑TEAAHB	29.9±0.10b	33.3±0.09a	33.16±0.08a

下载: 导出CSV

表 8 HP1胞外多糖后对乌鳢肌肉脂肪酸组成的影响

Table 8. Effect of HP1 exopolysaccharides on fatty acids in O. argus muscles

(单位/%)
脂肪酸 Fatty acid	C1	C2	C3
十五烷酸C15:0 Pentadecylic acid C15:0	0.34±0.01	0.32±0.02	0.31±0.02
棕榈酸C16:0 Cetylic acid C16:0	23.23±0.27a	22.47±0.36b	22.09±0.48b
棕榈烯酸C16:1 Palmitoleic acid C16:1	3.70±0.05	3.55±0.18	3.56±0.13
十七烷酸C17:0 Daturic acid C17:0	0.34±0.02	0.32±0.02	0.31±0.03
硬脂酸C18:0 Octadecanoic acid C18:0	5.27±0.24	4.95±0.17	5.13±0.21
反式油酸C18:1T Trans oleic acid C18:1T	0.11±0.01	0.11±0.02	―
顺式油酸C18:1 Cis oleic acid C18:1T	32.44±0.67a	31.61±0.85a	30.05±0.58b
顺式亚油酸C18:2 Cis linoleic acid C18:2	24.24±0.58	23.66±0.62	23.56±0.76
α亚麻酸C18:3N3 α-linolenic acid C18:3N3	0.22±0.02	0.32±0.01	0.31±0.01
γ亚麻酸C18:3n6 γ-linolenic acid C18:3n6	2.47±0.36	2.26±0.43	2.30±0.56
花生酸C20:0 Arachic acid C20:0	0.34±0.02	0.32±0.01	0.31±0.02
花生一烯酸C20:1 Cis-11-Eicosenoicacid C20:1	1.91±0.13a	1.94±0.09a	1.68±0.08b
二十一烷酸C21:0 n-heneicosanoic acid C21:0	0.56±0.01	0.65±0.05	0.52±0.02
γ-花生三烯酸C20:3N6 γ-Carbonium C20:3N6	0.11±0.01	0.11±0.01	―
花生四烯酸C20:4N6 Arachidonic acid C20:4N6	0.56±0.04	0.43±0.08	0.52±0.01
γ-花生三烯酸C20:3N3 γ-Carbonium C20:3N3	0.22±0.01b	0.43±0.02a	―
EPA C20:5 Eicosapentaenoicacid C20:5	1.01±0.25	0.75±0.12	1.05±0.23
DHA Docosahexaenoic acid C20:5	5.61±0.21b	4.95±0.28c	6.28±0.16a
EPA+DHA	6.62±0.23b	5.70±0.19c	7.33±0.18a
饱和脂肪酸Saturated fatty acid	30.08±0.13a	29.03±0.16b	28.67±0.18c
单不饱和脂肪酸Monounsaturated fatty acid	38.16±0.21a	37.21±0.29b	35.29±0.26c
多不饱和脂肪酸Polyunsaturated fatty acid	26.93±0.32a	26.24±0.35b	26.17±0.23b
高不饱和脂肪酸Highly unsaturated fatty acid	7.51±0.1b	6.67±0.11c	7.85±0.13a
n-3	7.07±0.12b	6.45±0.11c	7.64±0.13a
n-6	3.14±0.14a	2.80±0.17b	2.82±0.29b
n-3:n-6	2.25	2.31	2.71

下载: 导出CSV

参考文献(20)

[1]	李小勤, 胡斌, 冷向军, 等.VE对草鱼成鱼肌肉品质和抗氧化性能的影响[J].水生生物学报, 2009, 33(6):1132-1139. http://d.old.wanfangdata.com.cn/Periodical/ssswxb200906018 LI X Q, HU B, LENG X J, et al. Effect of supplemental VE on meat quality and anti-oxidant capacity of adult grass carp[J]. ActaHydrobiologicaSinica, 2009, 33(6):1132-1139.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/ssswxb200906018
[2]	王清滨, 王秋举, 杨翼羽, 等.牛磺酸对投喂高脂饲料草鱼幼鱼生长、肌肉品质及抗氧化能力的影响[J].西北农林科技大学学报(自然科学版), 2015, 43(7):49-65. http://d.old.wanfangdata.com.cn/Periodical/xbnydxxb201507008 WANG Q B, WANG Q J, YANG Y Y, et al. Effect of taurine on growth, muscle quality and antioxidant capacity of juvenile grass carp (Ctenopharyngodonidella) fed with high-fat diets[J]. Journal of Northwest A & F University (Natural Science Edition), 2015, 43(7):49-65.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/xbnydxxb201507008
[3]	王吉桥, 祁彩霞, 程爱香, 等.黄芪、熟地和山楂等中草药对黄颡鱼生长和消化的影响[J].水产学杂志2008, 21(1):34-41. http://d.old.wanfangdata.com.cn/Periodical/scxzz200801006 WANG J Q, QI C X, CHENG A X, et al. Effects of astragalus, ripe and hawthorn on the growth and digestibility of juvenile yellow catfish[J]. Chinese Journal of Fisheries, 2008, 21(1):34-41.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/scxzz200801006
[4]	向枭, 陈建, 周兴华, 等.黄芪多糖对齐口裂腹鱼生长、体组成和免疫指标的影响[J].水生生物学报, 2011, 35(2):291-299. http://d.old.wanfangdata.com.cn/Periodical/ssswxb201102013 XIANG X, CHEN J, ZHOU X H, et al. Effect of astragalus polysaccharides on growth, body composition and immune index in SchizothoraxPrenanti[J]. Acta Hydrobiologica Sinica, 2011, 35(2):291-299.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/ssswxb201102013
[5]	吴旋.四种中草药多糖对黄颡鱼生长、体成分及部分生理生化指标的影响[D].天津: 天津农学院, 2011. http://cdmd.cnki.com.cn/Article/CDMD-10061-1011191695.htm WU X. Effects of four kinds of Chinese herbal polysaccharides on the growth performance, body composition and part of physiological and biochemical indicator in yellow catfish (Pelteobagrus fulvidraco)[D]. Tianjin: Tianjin Agriculture University, 2011.(in Chinese) http://cdmd.cnki.com.cn/Article/CDMD-10061-1011191695.htm
[6]	袁桂秀, 林晨, 江振友, 等.改良焦性没食子酸法培养厌氧菌[J].暨南大学学报, 1999, 20(4):24-25. http://www.cnki.com.cn/Article/CJFDTOTAL-JNDX199904006.htm YUAN G X, LIN C, JIANG Z Y, et al. Cultivating anaerobic bacteria by modifying pyrogallic acid method[J]. Journal of Jinan University (Medicine Edition), 1999, 20(4):24-25.(in Chinese) http://www.cnki.com.cn/Article/CJFDTOTAL-JNDX199904006.htm
[7]	刘国红, 刘波, 陈峥, 等.一株产低温蛋白酶芽孢杆菌FJAT-24893的筛选与鉴定[J].福建农业学报, 2015, 30(10):954-957. http://www.fjnyxb.cn/CN/Y2015/V30/I10/954 LIU G H, LIU B, CHEN Z, et al. Screening and phylogenetic analysis on a protease-producing Bacillus species[J]. Fujian Journal of Agricultural Sciences, 2015, 30(10):954-957.(in Chinese) http://www.fjnyxb.cn/CN/Y2015/V30/I10/954
[8]	RAZA W, WU H S, SHEN Q.R. Use of response surface methodology to evaluate the effect of metal ions (Ca²⁺, Ni²⁺, Mn²⁺, Cu²⁺) on production of antifungal compounds by Paenibacilluspolymyxa[J]. Bioresource Technology, 2010, 101:1904-1912. doi: 10.1016/j.biortech.2009.10.029
[9]	郭振.改变水体盐度对吉富罗非鱼肉质的影响[D].天津: 天津农学院, 2014, 13-14. http://cdmd.cnki.com.cn/Article/CDMD-10061-1014225318.htm GUO Z. The effects of salinity changes on the muscle quality of Oreochromisniloticus[D]. Tianjin: Tianjin Agriculture University, 2014, 13-14.(in Chinese) http://cdmd.cnki.com.cn/Article/CDMD-10061-1014225318.htm
[10]	郑永标, 江枝和, 李三暑, 等. 868发酵液提取物对甲鱼白细胞吞噬能力的影响[J].福建农业学报, 2001, 2(16):43-45. http://www.fjnyxb.cn/CN/Y2001/V/I2/43 ZHENG Y B, JIANG Z H, LI S S, et al. Effect of 868 extraction on phagocytosis of leucocyte of Trionyxsinensis[J]. Fujian Journal of Agricultural Sciences, 2001, 2(16):43-45.(in Chinese) http://www.fjnyxb.cn/CN/Y2001/V/I2/43
[11]	RAVI K P, PAPORI G, ABU T K, et al. Structural analysis and properties of dextran produced by Leuconostocmesenteroides NRRL B-640[J]. Carbohydrate Polymers, 2009, 76:30-35. doi: 10.1016/j.carbpol.2008.09.018
[12]	LU Z Q, JIN M L, HUANG M, et al.Bioactivity of selenium-enriched exopolysaccharides produced by Enterobacter cloacaeZ0206 in broilers[J]. Carbohydrate Polymers, 2013, 96:131-136. doi: 10.1016/j.carbpol.2013.03.063
[13]	REHFELDT C, FIEDLER I, DIETL G, et al. Myogenesis and postnatal skeletal cell growth as influenced by selection[J]. Livestock Production Science, 2000, 66(2):177-188. doi: 10.1016/S0301-6226(00)00225-6
[14]	石英, 冷向军, 李小勤.杜仲叶提取物对异育银鲫生长、血清非特异性免疫和肌肉品质的影响[J].浙江大学学报(农业与生命科学版), 2008, 34(2):200-206. http://d.old.wanfangdata.com.cn/Periodical/zjdxxb-nyysm200802013 SHI Y, LENG X J, LI X Q, et al. Effect of eucommiaulmoidesoliver leaf extract on growth, serum non-specific immune response and meat quality of crucian carp[J]. Journal of Zhejiang University (Agric & Life Science Edition), 2008, 34(2):200-206.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/zjdxxb-nyysm200802013
[15]	舒琥, 周莹, 肖丹, 等.野生三斑石斑鱼肌肉营养成分与品质的评价[J].安徽农业科学, 2011, 39(21):12902-12905. http://d.old.wanfangdata.com.cn/Periodical/ahnykx201121096 SHU H, ZHOU Y, XIAO D, et al. Evaluation of nutrient components and nutritive quality in muscle of wildEpinephelustrimaculatus[J]. Journal of Anhui Agricultural Science, 2011, 39(21):12902-12905.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/ahnykx201121096
[16]	邴旭文, 蔡宝玉, 王利平.中华倒刺鲃的肌肉营养成分与品质的评价[J].中国水产科学, 2005, 12(2):211-215. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=zgsckx200502018 BING X W, CAI B Y, WANG L P. Evaluation of nutritive quality and nutritional components in Spinibarbussinensismuscle[J]. Journal of Fishery Sciences of China, 2005, 12(2):211-215.(in Chinese) http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=zgsckx200502018
[17]	胡兵, 刘军, 侯永清, 等.黄芪多糖对异育银鲫肌肉组成成分的影响[J].中国饲料, 2008(11):37-44. http://d.old.wanfangdata.com.cn/Periodical/zgsl200811017 HU B, LIU J, HOU Y Q, et al. Effect of astragalus polysaccharides on nutritional ingredient in muscle of carassiusauratusgibelio[J]. China Feed, 2008(11):37-44.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/zgsl200811017
[18]	李卓佳, 林黑着, 郭志勋.中草药对斑节对虾生长、饲料利用和肌肉营养成分的影响[J].南方水产, 2007, 3(2):20-24. http://d.old.wanfangdata.com.cn/Periodical/nfsc200702004 LI Z J, LIN H Z, GUO Z X. Effect of traditional Chinese medicines on growth, feed utilization and biochemical composition in muscle of juvenile shrimp Penaeusmonodon[J]. South China Fisheries Science, 2007, 3(2):20-24.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/nfsc200702004
[19]	王树海, 王文健, 汪雪峰, 等.黄芪多糖和小檗碱对3T3-L1脂肪细胞糖代谢及细胞分化的影响[J].中国中西医结合杂志, 2004, 24(10):926-928. http://d.old.wanfangdata.com.cn/Periodical/zxyjh200410017 WANG S H, WANG W J, WANG X F, et al. Effect of astragalus polysaccharides and berberine on carbohydrate metabolism and cell differentiation in 3T3-L1 adipocytes[J]. Chinese Journal of Integrated Traditional and Western Medicine, 2004, 24(10):926-928.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/zxyjh200410017
[20]	王苗苗, 王海磊, 罗庆华.鳡鱼肌肉营养成分测定及评价[J].食品科学, 2014, 35(15):238-242. http://d.old.wanfangdata.com.cn/Periodical/spkx201415048 WANG M M, WANG H L, LUO Q H, et al. Analysis and evaluation of nutrient components in muscle of yellow cheek carp[J]. Food Science, 2014, 35(15):238-242.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/spkx201415048