Complementary acupionts (CAs)[1-2], refer to the two acupoints respectively representing yin and yang,located in the head, neck, trunk, limbs, and other body parts, selected at the same time to play a synergistic effect. It’s characterized by treating yin for yang disorders and treating yang for yin disorders[3]. CAs are convenient for clinical application with good therapeutic effect[4-5]. Mast cells (MCs) are correlated with the meridian phenomenon, and play an important role in stimulation and regulation during acupuncture.Mechanical stimulation can degranulate MCs to release many active substances, such as bradykinin (BK),histamine (HIS), and prostaglandin (PG)[6-8]. The MCs stimulate multi-targeted cellular network communication by mobilizing, recruiting, degranulation and release of multiple active substances. Although CAs show satisfactory clinical efficacy, the synergistic mechanism is not clear[9].

In this study, electroacupuncture (EA) was performed to stimulate the CAs, Yinlingquan (SP 9) and Yanglingquan (GB 34). The MCs and the changes in released HIS and BK were observed to explore the basic mechanism of synergistic action of CAs, thus to provide ideas and basis for profound study and clinical application of the combination theory of CAs.

1 Materials and Methods

1.1 Laboratory animals and groups

Forty healthy female Wsitar rats, weighing 190-200 g,were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. During the experiment, the processing of animals completely conformed to the Guiding Opinions on the Treatment of Experimental Animals.

3.2.2 分时段收取停车费用。适当调节停车需求，根据停车时间的长短进行分时段、分地段收费，可以实行弹性收费制度，鼓励短时停车以提高停车位的周转率。在此基础上，也可以对有不同需求的群体进行不同的收费标准，对长时间占用车位的人适当提高收费标准，从而确保停车场的利用处于相对均衡的状态。

Grouping methods: Using random number table method, the rats were randomly divided into an acupuncture at Yanglingquan (GB 34) group (group A),an acupuncture at Yinlingquan (SP 9) group (group B),an acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group AB), an acupuncture at points beside Yanglingquan (GB 34) and Yinlingquan(SP 9) group (group CD) and a normal control group(group K), 8 rats in each group.

1.2 Instruments and reagents

At the same time, it is suggested that the changes in MCs recruitment and degranulation in tissues of the acupoints and the changes in serum HIS and BK levels can be used as reference indicators to quantify the acupuncture effect, and provide some experimental and theoretical basis for the clinical evaluation of acupuncture efficacy.

1.3 Experimental methods

HIS is a vasoactive amine compound in the tissues of human body. It exists widely in the tissues of the human body and is highest in MCs. HIS is also present in the tissues of skin, peripheral nerves, lungs, liver and gastric mucosa. In most cases, HIS is released by MCs after trauma or external specific antigen stimulation. Studies have shown that, HIS in the peripheral nerve is a pain-inducing substance that can cause physical pain and antagonist of acupuncture analgesia; in the central nervous system, it is an alleviation that plays a role in analgesic effect and synergetic acupuncture analgesia[17].Previous studies have shown that the 5-HT or HIS was positive in basophilic granules released from MCs in tissues of the acupoints after stimulation by acupuncture and moxibustion[18].

小型水库数量较多的乡镇 （街道），建立健全水库管理组织，对辖区内乡镇（街道）所有和农村集体经济组织所属的小型水库实行区域化集中管理。水库工程日常维修养护即可委托水利工程维修养护公司，也可由水库管理单位自行组织实施。

1.4.1 Serum HIS and BK levels

Yinlingquan (SP 9): On the medial side of the cruris,opposite to Yanglingquan (GB 34).

Group A: Acupuncture at bilateral Yanglingquan(GB 34).

Group B: Acupuncture at bilateral Yinlingquan (SP 9).Group AB: Acupuncture at bilateral Yanglingquan(GB 34) and Yinlingquan (SP 9).

Specimens from rats in each group were collected 2 h after the 7th day. Hair around the CAs was shaved,firstly with an electric hair clipper, and then with an electric shaver. The skin (about 1 cm × 1 cm) was marked with the acupoint as the center. The anesthetized rats using intraperitoneal injection of 3.5%chloral hydrate [10 mL/(kg·bw)] were sacrificed by dislocation after blood collection via abdominal aorta.

Group CD: Points 3 mm away from bilateral Yanglingquan (GB 34) and Yinlingquan (SP 9) were used as control points and subjected to acupuncture[11].

Group K: Rats were fixed using the same method as rats in the other 4 groups without acupuncture stimulation.

Needles of 0.35 mm in the diameter and 40 mm in length were used for acupuncture. The handles were connected to the G6805-Ⅱ mode EA device after needling qi was obtained. Sparse-dense wave,frequency of 2 Hz/100 Hz and current of 1 mA were used to keep the needle handles slightly tremulous while the rats kept quiet. Rats were continuously stimulated by EA for 20 min each time. Experimental interventions were conducted on the 1st, 3rd, 5th and 7th days when the experiment started, for 4 times in total. Rats were housed as normal overnight after each intervention. All acupuncture interventions were conducted by trained professionals started from 3 p.m.The intervention order was group A, group B, group AB,group CD and group K.

1.4 Observation items and testing methods

胡四一：最严格水资源管理制度控制指标的确定，既考虑了我国国情和水情，也参照了水资源可持续利用的国际先进水平。所谓最严格的水资源管理目标，实际体现在“三条红线”上。“红”就意味着是最严格，“线”是一个管控的目标，最严格的水资源管理制度的核心实际上是最严格的水资源管控目标。“三条红线”是我们水资源开发利用的一个底线，一旦突破这样的底线，经济社会发展就要受损，生态环境就要受到严重影响。

Yanglingquan (GB 34): Five millimeter above and lateral to Zusanli (ST 36).

The abdominal aorta blood was harvested and centrifuged at 3 500 r/min immediately after anesthesia.The serum was collected and the levels of HIS and BK were determined by ELISA according to the kit instruction manual.

1.4.2 The number and the degranulation rate of MCs in tissues of the acupoint regions

Tissues (including the skin, subcutaneous tissue and a small amount of muscle, about 1 cm × 1 cm × 0.4 cm) of the acupoint regions were collected. The skin and subcutaneous tissues of the acupoint regions were collected in group A and group B; the skin and subcutaneous tissues of Yinlingquan (SP 9),Yanglingquan (GB 34) and those between the two acupoints were collected in group K, group AB and group CD. The collected tissues were wrapped with foil,put into the labeled frozen tube, and stored at –80 ℃refrigerator. Sections (thickness of 15 μm) were sliced in a constant cold box for immune-histochemistry staining against acetylcholinesterase with TBO counterstaining[12-13], followed by a gradient of alcohol dehydration, xylene transparent and neutral gum seal.Four sections were randomly selected for each rat, and 3 fields were observed for each section under optical microscope (× 100). The number of MCs and number of degranulated MCs were recorded in each field.

Statistics of the MCs numbers and degranulated MCs numbers: The MCs number for each slice was the total number of MCs in 3 fields per slice, and the average MCs number of the 4 slices was designated as the MCs number of each rat.

Degranulation rate = Number of degranulated MCs ÷MCs number × 100%.

1.5 Statistical method

Data were analyzed using SPSS 19.0 version statistical analysis software. Measurement data were presented as mean ± standard deviation (x±s), and the differences between groups were compared using one-way analysis of variance. If the variance was homogeneous, the least significant difference (LSD)method was used for comparison; if the variance was heterogeneous, then the Tamhane method was used for comparison. P＜0.05 indicated that the difference was statistically significant.

2 Results

2.1 General conditions of the rats

Before the intervention, the rat’s body weight in each group was 190-200 g and showed no statistically significant difference among groups (P=0.41), indicating the comparability. After the intervention, the rat’s body weight in each group was 200-220 g and showed no statistically significant difference among groups (P=0.89),(Table 1).

Table 1. Changes in rat’s body weight of each group (x±s, g)

Group n Before intervention After intervention Group K 8 192.33±2.08 209.00±4.58 Group A 8 194.00±2.35 211.60±3.13 Group B 8 195.40±2.07 209.80±3.77 Group AB 8 195.00±2.74 210.20±4.44 Group CD 8 195.00±2.00 210.60±3.05

2.2 Number and degranulation of MCs in tissues of the acupoints

The morphological observation of tissue sections in this experiment showed that different sizes and different numbers of MCs were distributed in dermis,subcutaneous tissue and muscle of the acupoints in each group (Figure 1). The shape of MCs was regular,mostly round or oval; the cell membrane was integral;the cytoplasm of MCs was filled with heterogenous particles; rare cells around showed scattered blue particles; degranulation was not obvious, around acupoints in rats of group K and group CD. In group A, B,and AB, the numbers and the volumes of MCs were increased; the cell edges were irregular; the cell membrane became thinner and ruptured; and a large number of blue particles were distributed around the cells with obvious degranulation.

Data statistics: Compared with group K, the MCs numbers in group AB, group A, group B and group CD were significantly increased (all P＜0.05), which was highest in group AB and statistically significantly different from the other groups (all P＜0.05). MCs numbers showed no statistically significant difference between group A and group B (P＞0.05). Compared to group K, the MCs degranulation rates of the other 4 groups all increased (all P＜0.05). MCs degranulation rate of group AB was the highest, there were statistical differences between group AB and the other 3 groups(group A, group B and group CD); MCs degranulation rate was not significantly different between group A and group B (P＞0.05), nor between group CD and group K(Figure 2).

Figure 1. Morphological structure and distribution of MCs in the tissues of the acupoints in each group (TBO staining, × 100)

Figure 2. MCs number and degranulation rate in tissues of the acupoints in each group

Note: Compared with group K, 1) P＜0.05; compared with group A, 2) P＜0.05; compared with group B, 3) P＜0.05; compared with group CD, 4) P＜0.05

2.3 Comparison of histamine and bradykinin levels

The order from high to low of histamine level in rats’serum: group AB ＞ group B ＞ group A ＞ group CD ＞group K; the bradykinin level in group AB was significantly higher than those in the other groups (all P＜0.05); the difference in bradykinin level between group A and group B was not statistically significant(P＞0.05); the difference in bradykinin level between group CD and group K was not statistically significant(P＞0.05), (Table 2).

Table 2. Comparison of histamine and bradykinin levels in rat’s serum of each group (x±s, ng/mL)

Note: Compared with group K, 1) P＜0.05; compared with group A, 2) P＜0.05; compared with group B, 3) P＜0.05; compared with group CD, 4) P＜0.05

Group n Histamine Bradykinin Group K 8 25.99±4.57 772.55±83.75 Group A 8 53.13±3.301) 925.16±88.861)Group B 8 56.95±9.011)2) 941.86±67.551)Group AB 8 69.15±9.801)2)3)4) 1 038.06±57.911)2)3)4)Group CD 8 35.85±5.291)2)3) 776.17±32.442)3)

3 Discussion

Human body consists of yin and yang, thus the pathogenesis is nothing more than imbalance between yin and yang (excess or deficiency). ‘Complementary acupiont’ is a new theory of piont combination based on the meridian pathways, the relationship between meridians and points, meridian theory, especially the yin and yang theory[3]. Yinlingquan (SP 9) and Yanglingquan (GB 34) are relative in yin and yang, and a pair of CAs widely used clinically[14]. Therefore,Yanglingquan (GB 34) and Yinlingquan (SP 9) were selected to study.

It has been proven that MCs are involved in the initiation and conduction of acupuncture effect information[15], thus are one of the key factors in acupuncture signal cascade amplification. After acupuncture stimulation, the MCs in the tissues ofthe acupoints are activated and degranulated to release various active substances such as HIS, BK,5-hydroxytryptamine (5-HT), PG and substance P (SP),which transform the physical stimulation of acupuncture into biological information to produce acupuncture effect, through the conduction and integration of the complex systems of nerve, endocrine,immune network, thus to play a regulatory role in target organs[16]. Therefore, the MCs number and degranulation rate, and the levels of histamine and bradykinin released by MCs in tissues of the acupoints were selected to observe the changes of body after acupuncture at the CAs.

Groups A, B, AB and CD: After the rats were fixed, the acupoints of rats were located according to the Experimental Acupuncture Science[10]. Yanglingquan(GB 34) and Yinlingquan (SP 9) were selected, and the acupuncture sites were marked with picric acid. i

赵永金[1]对于影响清代帖学式微因素的探讨比较全面。他认为，早在清初碑派尚未形成气候之时，帖学已呈现出衰微之势，考其原因主要有5个方面：①经典作品价值取向的转移；②对近世名家的盲目崇拜；③刻帖泛滥与金石学复兴对取法的影响；④书法教育与品评的缺失；⑤清早期书论的悖误。这5个因素无疑都影响了帖学的发展。这些因素出现在清初书坛，决不是偶然的巧合，而是当时社会环境、当时人们的观念意识和审美理想在艺术活动中社会心理情绪的真实反映，包涵着深刻的历史因素。

当新赛季开打前一个多月，吴庆龙接过凯撒的教鞭时，整个夏训已经结束，球队的外援组合、内援引进没有丝毫变化余地。名义上出资人、俱乐部和主教练都换了，但球队残缺不全的人员结构和糟糕的备战状态却是从前任那里全盘接收过来的，西王和吴庆龙注定要为高速和凯撒还债。

Conflict of Interest

The results of this experiment showed that acupuncture could increase local MCs recruitment and degranulation rate, which was most significant after acupuncture at CAs of Yinlingquan (SP 9) and Yanglingquan (GB 34), and thus confirmed the‘amplifier’ of acupuncture effect[20]. These results were in line with the previous results and experimental hypotheses. Acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9) group was significantly higher than acupuncture at Yanglingquan (GB 34) group,acupuncture at Yinlingquan (SP 9) group, and acupuncture at points beside Yanglingquan (GB 34) and Yinlingquan (SP 9) group, indicating that the acupuncture effect in acupuncture at CAs group was better than acupuncture at single acupoint and non-acupoints. Therefore, this experiment basically concluded that the increase of MCs recruitment and degranulation rate was one of the possible factors for the synergism of CAs combination. The statistical results showed that the serum levels of HIS and BK in rats after acupuncture were increased, which showed a positive correlation between them; and the serum histamine level reached the highest after acupuncture at CAs of Yinlingquan (SP 9) and Yanglingquan (GB 34). It is basically considered that the increase of serum HIS and BK may be one of the reasons for the synergy of CAs.

In this experiment, we found that the changes in morphology and recruitment trends of MCs, HIS and BK levels were related to the synergistic features of CAs.The synergism of acupuncture at CAs was confirmed via experimental study, which provided a new idea and basis for further exploring the mechanism of synergism,and the wider clinical application of CAs.

推荐理由:《疯狗浪》是一部感人至深的动物小说，讲述小狗沫沫与主人船花、大公狗黑风之间的故事，作品以动物的名义重新诠释“爱”，带给孩子们直击心灵的感动。这部作品再次刷新了曹文轩的文学高度和文学成就，以崭新的面貌面向读者，带给读者一个不一样的曹文轩。

Thioacetylcholine (Tokyo Chemical Industry, Japan);toluidine blue O (TBO, AMRESCO, USA); cryostat slicer(Thermo Fisher Scientific, UK); Multiskan MK3 enzyme analyzer (Thermo, Finland); enzyme-linked immunosorbent assay (ELISA) kit (Shanghai Enzyme Biotech Co., Ltd., China); G6805-Ⅱ EA instrument(Qingdao Xin Sheng Industrial Co., Ltd., China).

BK is an essential inflammatory mediator, converted from prokinin under the action of kallikrein after external irritant. The role of BK in the body is extremely broad. On the one hand, it can sensitize the sensory neurons and trigger the body's inflammatory response;on the other hand, it also involves in glandular secretion,smooth muscle contraction and other physiological activities. BK has played an integral role in the production and conduction of pain, and acupuncture analgesic process[19].

The authors declared that there was no potential conflict of interest in this article.

上面两例中，句子后面括号里的动词可以出现，也可以省略。在描写语体中，很多动词都可以省略，形成了所谓的体词性谓语句。当然体词性谓语句也是很复杂的一个句型范畴，并不仅仅完成描写语体功能，还可以有其他的功能，比如“鲁迅绍兴人”、“今天星期一”等表示处所和时间的判断句。但我们注意到典型的动词谓语句一般不属于描写语体，如上文例(1)(2)中的核心动词，是不能省略的。

建立增减挂钩节余指标储备机制的实践和思考（王志卿） .............................................................................3-35

This work was supported by Project of Natural Science Foundation of Hebei Province (河北省自然科学基金项目, No. H2014406047); Key Program of Science and Technology of Hebei Higher Education (河北省高等学校科学技术研究重点项目, No. ZD20131001).

高温颜色釉艺术作品的主要艺术形式表现为色彩的多样式，颜色釉陶艺作品都是经过高温烧制而成，在窑炉里产生极具天然性的肌理效果，由于高温颜色釉作品制作难度较大，烧坏率比较高，烧制出来的效果又极具特色，而且每次烧制的效果都不一样，因此，作品极具偶然性，有种可遇不可求的特性。

Statement of Human and Animal Rights

The treatment of animals conformed to the ethical criteria in this experiment.

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