Supported by Fundamental Study on Rules and Influencing Factors of Specificity of Acupoint Effect (2012CB518502).

7890A-5975C气相色谱-质谱联用仪，美国Agilent公司；Nicolet 380傅里叶变换红外光谱仪，美国Thermo公司；UV-2550双光束紫外光谱仪，日本岛津公司；SGZ-B系列便携式浊度计，上海悦丰仪器仪表有限公司；SCIENTZ-18N真空冷冻干燥机，宁波新芝生物科技股份有限公司；低速离心机，安徽中科中佳科学仪器有限公司；比色管，天津玻璃仪器有限公司；1 cm密封石英比色皿，德国莱斯公司。

1 Introduction

According to studies of Mu Xiang etal., intradermal microvascular groups in the acupoint area have synchronous vasomotion phenomenon, significantly different from the non-acupoint area; on this basis, they stated that the essence of acupoint is the microvascular groups with synchronous vasomotion function within the skin in the acupoint area[1]; further study indicated that acupuncture can significantly increase the amplitude of microvascular vasomotion within the skin in the acupoint area[2], increasing the amplitude of microvascular vasomotion in the lesion area can significantly improve the therapeutic effect of acupuncture on intervertebral disc disease (IVDD) of dogs[3]. However, to perform acupuncture, it is necessary to fully understand the specific acupoint, grasp acupuncture skills, which is clinically difficult. In YellowEmperor’sInnerCanon, there is record that acupuncture and moxibustion are often combined with herbs. Acupoint administration effectively combines acupuncture and moxibustion and has been widely used in clinical practice since the ancient times. Acupoint can be used as drug administration window, absorbing drugs and taking a therapeutic effect. Previous studies of our laboratory have proved that the introduction of ginsenoside Rb1 by the iontophoresis device can effectively increase the amplitude of microvascular vasomotion in acupoint area. Therefore, we try to prepare ginsenoside Rb1 Babu agent, and explore effects of ginsenoside Rb1 Babu agent on amplitude of microvascular vasomotion, so as to develop a topical drug patch that can replace the acupuncture treatment.

第二种迹象：微小的日常。一些艺术家已经抛开了所谓宏大叙述，转向了更加贴近自身的、细微的日常景观，展开更加深入、细腻的表达，提供了前所未有的独特视角。

2 Materials

2.1 Medicines and reagents Ginsenoside Rb1 (Shanghai Tauto Biotech Co., Ltd., CAS: 41753-43-9); sodium polyacrylate (batch number: S108369); sodium carboxymethyl cellulose (batch number: C104986), Arabic gum (batch number: A108975), gelatin (batch number: G108396), kaolin (batch number: 38120), lanolin (batch number: L116310), glycerol (batch number: G116203), azone (batch number: A104374), bought from Aladdin Reagent (Shanghai) Co., Ltd.); anhydrous ethanol (Beijing Chemical Works, batch number: 20090601); saline injection (Hebei Tiancheng Pharmaceutical Co.,Ltd., batch number: 090710116).

2.2 Instruments Intelligent transdermal apparatus (Tianjin Jingtuo Instrument Sci & Tech Co., Ltd.); PeriFlux System 5000 Laser Doppler Flowmeter (Sweden Perimed AB); PROBE407 laser probe, PH07-4 probe holder, PF 105-3 adhesive (Sweden Perimed AB); WQ6-F30 acupuncture meridian locator (Donghua Electronic Instrument Factory in Haidian District, Beijing); UT39 series of new digital multimeter (Uni-Trend Electronic (ShangHai) Limited of China); Boyu BY3005S single DC steady power supply (Beijing Boyu Xunming Technology Co., Ltd.).

where R iscumulativepermeationrate (%); Cnand Cidenotethedrugconcentration (μg/mL) measuredatthe n and i samplingpointsrespectively; V and Videnotethevolume (mL) ofdiffusioncellandthe i-thtimesampling; m isthetotalmassofthedrug (μg). Takingthecumulativepermeationrate (R) asordinateandthetime (t) asabscissa, weplottedachartandobtainedthecumulativepermeationrate-timecurve, asshowninFig.5. Withtheextensionoftransdermaltime, thecumulativepermeationrateofginsenosideRblincreased.

农村产业融合是一场新的产业技术革命，是中国农业发展的必然选择[1]。近年，东营市作为国家级现代农业示范区，农业农村经济发展方式转变迅速推进，农业产业化集群初具规模，农村一、二、三产业融合发展。但是，一、二、三产业的融合发展遇到了许多问题和矛盾，亟需解决。加快东营市农村一、二、三产业的融合发展势在必行、刻不容缓。

2.4 Experimental subjects Eight 20-30 year-old healthy volunteers.

3 Preparation of Babu agent

3.1 Preparation of Babu agent containing drugs Precisely weighed 0.75 g of sodium polyacrylate, 0.3 g of sodium carboxymethyl cellulose, 7.5 g of Arabic gum, and 7.5 g of gelatin, mixed them well and dispersed in water with 20 mL formula amount and swelled for 3 h. At the same time, dispersed 3 g of kaolin in 19 mL of glycerin and mixed well. Thereafter, in a boiling water bath, stirred the mixture of the swollen sodium polyacrylate, and added the glycerin mixed with kaolin and 20 mL of water and 1 g of lanolin, and stirred for 20 min. The resulting clear gel was the desired Babu agent matrix.

Previous experiment has proved that when using 5% azone as a transdermal enhancer, the permeation of drug in this Babu agent was optimal. Selected 0.5% drug concentration. Specifically, weighed 25 mg of ginsenoside Rb1, added it into the mixture of 1 mL of ethanol and 0.25 mL azone, after it was fully dissolved, added 5 g matrix that has been dissolved in water bath; after well mixing, dispersed it on an acupoint blank patch, cooled down, and stored at 4℃ for use.

模拟小试结果初步验证了植物能够在掺混有机肥料的底泥中生长良好。因此，在江西德兴铜矿水龙山排土场选择10000m2的区域进行了中试试验。按模拟小试最佳结果，底泥控制在体积30%与有机肥料配比，并与排土场表层30cm的土壤掺混，进行土壤改良。按2株/m2的用量直栽乔灌木袋苗与撒播植物种子、土壤种子库，种植完成后，用稻草覆盖遮荫。中试试验取得了良好生态恢复效果，进一步证实底泥添加有机肥料调配制成的人工基质能够满足植被重建的需求（见图2）。

Before the experiment, started the Laser Doppler Flowmeter, connected the probe, and preheated for 20 min. Fixed the Probe 407 of the Laser Doppler Flowmeter onto the acupoint, and it should be noted that the probe should be perpendicular to the area to be tested. When the changes of graphic movement became stabilized, we started recording. After the first test was over for 3 d, proceeded to the next test.

4 Theinvitro transdermal permeation of ginsenoside Rbl Babu agent

4.1 Preparation ofinvitro mouse skin Selected Kunming variety male mice with body weight of 18-22 g, removed abdomen hair with the hair removal cream, fed one day; before experiment, killed through dislocating the cervical spine, fixed, took the abdomen skin, removed the subcutaneous fat, capillaries and mucosa, selected intact skin, cleaned repeatedly with saline, put into the refrigerator at low temperature for use.

4.2 Transdermal permeation test The test adopted Franz diffusion device (the volume of receiving chamber volume was 15 mL, effective contact area was 0.785 cm2); added 37℃ preheated saline in the receiving chamber, made the skin epidermis face inward and corneum face outward, pasted Babu agent outside the corneum, fixed the device, conducted the test at constant temperature of 37℃ and stirring speed of 300 r/min, took samples at 0.5, 1, 2, 3, and 4 h, each time 2 mL, and supplemented with the same volume of saline. Then, filtered the collected samples by 0.22 μm microporous membrane, and carried out high performance liquid chromatography (HPLC) analysis to detect the drug transdermal permeation.

4.3 HPLC analysis of percutaneous fluid Chromatographic conditions for ginsenoside Rb1 detection: chromatographic column (Waters C18,150 mm × 4.6 mm, 5 μm), the column temperature of 25℃, the flow rate of 1 mL/min, the injection volume of 100 μL. The mobile phase was acetonitrile -0.1% phosphoric acid solution (34∶66) and the detection wavelength was 203 nm.

5 Detection of amplitude of microvascular vasomotion

In this study, we recruited eight healthy college students as experimental subjects. Each volunteer was applied with ginsenoside Rb1 Babu agent and blank patch, respectively, and recorded as drug group and control group.

5.1 Selection and positioning of acupoint In the process of experiment, ginsenoside Rb1 Babu agent has to be pasted to the acupoint. Therefore, we selected two acupoints (Quchi acupoint and Shousanli acupoint) with the same meridian. Quchi acupoint was used for pasting, and Shousanli acupoint was used for detection.

大数据给创新培养提供了积极的思路，但是与数据相联系，人们很容易就会联想到信息和数据安全问题。学生的信息一旦被泄露，将给他们的人身安全和日常生活带来极大威胁和困扰。如何确保学生的个人信息安全与隐私、如何防范数据资料不被商业化利用，是我们在对面教育类数据时不可回避的问题，同样是出于隐私和安全的考察，教育数据在很多高校内部通常被视作为需要保密的资料，教师和部门以外的人员要想获取数据存在困难。华中师范大学的一学者在开展一课题研究时，需要用到学生的图书借阅数据，最后是通过多方沟通及书面保证才顺利拿到数据。研究人员都能理解职能部门的这一规定，但在客观上也确实不利于开展大数据的研究。

这是我人生中第一次替父亲抽号。那是2011年的夏天，巨浪牧场2号楼竣工。农场场部机关会议室人声鼎沸、座无虚席。父亲早已激动得不知迈哪条腿走路，还是母亲比较淡定：“二楼好，更上一层楼嘛！”

5.1.2 Low resistance point color development method of acupoint. The position of acupoint to be detected was roughly determined according to the anatomical location, scrubbed with clean water, and dried. Gently placed the acupoint electrode (copper sheet with 20 mm diameter and 0.5 mm thickness) at the acupoint to be detected, connected with the negative pole of Boyu BY3005S single DC steady power supply, reference electrode (two stainless steel sheet with size of 40 mm × 20 mm and thickness of 0.2 mm) padded with saline soaked gauze clipped ears of experimental subjects, connected in series with UT39 new digital multimeter (2 mA grade). Switched on the power, observed the readings of digital multimeter, the current readings would gradually increase over time, the output voltage of power supply should be controlled at any time to control the current intensity. When the current reached 0.3 mA, started counting 60 s, cut off the power supply, applied the color developer at the acupoint electrode action area, then there would display the rose red dot. The position of this rose red dot was the acupoint.

5.2 Detection of amplitude of microvascular vasomotion The experimental subjects were eight healthy college students. Before the experiment, they should avoid any activity that may affect the skin perfusion, and should adapt to the environment for 20 min in a quiet room with temperature of 25℃. To avoid artifacts caused by the measurement of the target or body movement, subjects should keep quiet as much as possible during the experiment.

5.1.1 Preparation of acupoint developer. We boiled 100 mL of distilled water, add 9 g of methyl cellulose, stirred at the same time, added 0.8 g of phenolphthalein reagent, fully stirred, then added ethanol solution with 1 mL of 15% methyl paraben and 1 mL of 2.5% ethylparaben, and added 2 mL of 10% benzalkonium bromide, cooled down for use.

研究结果显示：与基础组相对比，分析组患者的不良反应率低，依从率和满意率高，图像质量优（P＜0.05）。原因分析：临床护理路径在现代护理理念基础上，继承传统护理优点，优化护理不足之处，围绕患者为中心，从病情、心理、生理等方面，设计护理方案，开展护理工作，有效确保临床疗效，促使护理质量提高，让患者更加满意护理服务。临床护理路径规范作为护理人员执行工作的标准，护理人员严格要求自己按照护理计划实施工作，有效弥补护理人员素质参差不齐、能力不足的缺陷，为患者提供更加优质、舒适的护理服务。

3.2 Preparation of blank Babu agent Used 5 g of blank matrix, 1 mL of ethanol and 0.25 mL of azone by the operation method in Section 3.1.

6 Image processing and data analysis

From each image for amplitude of microvascular vasomotion, selected five segments of data and calculated the changes in amplitude of acupoint. Changes in amplitude = PUmax-PUmin.

The data were processed by Prism statistical software; the measurement data were expressed as mean ± standard deviation (s); comparison between groups was analyzed by t test, P<0.05 denoted statistically significant difference, while P<0.01 denoted extremely statistically significant difference.

7 Experimental results

7.1 Results of theinvitro transdermal permeation of ginsenoside Rbl Babu agent The ginsenoside Rb1 standard product was analyzed by liquid phase, as shown in Fig.1. The ginsenoside Rb1 appeared at about 4.3 min. Invitro transdermal liquid-phase analysis of Babu agent not containing ginsenoside-free Rb1 was shown in Fig.2. According to Fig.2, Babu agent component did not interfere with the determination of ginsenoside Rb1. Through invitro transdermal liquid analysis of ginsenoside Rb1 Babu agent, the drug has permeated the skin at 0.5 h, as shown in Fig.3 and Fig.4.

Fig.1 Ginsenoside Rb1 standard product

Fig.2 Chromatogram for Babu agent not containing ginsenoside Rb1

Fig.3 Ginsenoside Rb1 permeated the skin at 0.5 h

Fig.4 Ginsenoside Rb1 permeated the skin at 1 h

The microvascular vasomotion is complex cyclical biological wave with unique amplitude and frequency[4]. In many diseases, such as diabetes, hypertension and obesity, the microvascular vasomotion declines. Previous studies have shown that acupuncture can significantly increase the amplitude of microvascular vasomotion within the skin in the acupoint area, thus increasing the amplitude of microvascular vasomotion is of potential significance for the treatment of diseases. However, the application of acupuncture requires professional staff and special tools, which brings many inconveniences to the patients. In addition, some patients may have a certain fear of acupuncture treatment, and some patients even faint during acupuncture. According to TCM theories, acupuncture and moxibustion are often combined with herbs, TCM treatment and acupuncture treatment mechanism should be the same. Acupoint can be used as drug administration position, absorbing drugs and taking a therapeutic effect. Acupoint administration effectively combines acupuncture and moxibustion and has been widely used in clinical practice since the ancient times[5-7].

2.3 Experimental animals Kunming variety male mice with body weight of 18-22 g (Beijing Xinglong Experimental Animals Co., Ltd.).

Fig.5 RelationshipbetweencumulativepermeationrateofginsenosideRb 1andtime

7.3 Detection results of amplitude of microvascular vasomotion We found that after the subjects pasting the ginsenoside Rb1 Babu agent, the amplitude of microvascular vasomotion was significantly increased (P<0.01). However, in the control group, after pasting the Babu patch, the amplitude of microvascular vasomotion of some subjects may also show significant increase (P<0.01), but the amplitude of increase was not significant as pasting the Babu patch containing drugs. Statistical charts for the changes in amplitude of microvascular vasomotion of some subjects were illustrated in Fig.6.

8 Discussions

7.2 Cumulative permeation rate of ginsenoside Rb1 Calculation formula for cumulative permeation rate at different time:

本组8例患者中，男6例，女2例；年龄28～62岁，平均（44.24±11.26）岁；高处坠落伤3例，交通伤5例。患者均有不同程度的疼痛和髋关节活动受限。所有患者术前行骨盆正位、入口位、出口位DR片及骨盆CT扫描三维重建检查并行患侧股骨髁上牵引。手术时间平均为伤后（12.32±1.44） d。

Fig.6 Changes in amplitude of microvascular vasomotion of some subjects

Ginsenoside Rb1 has excellent therapeutic effect on the central nervous system, cardiovascular system, and immune system and has anti-tumor, anti-liver heat ischemia-reperfusion injury, and hypoglycemic effect[8]. Previous studies have proved that the introduction of ginsenoside Rb1 by the iontophoresis device can effectively increase the amplitude of microvascular vasomotion in acupoint area[9], in other words, acupoint administration of ginsenoside Rb1 can achieve acupuncture effect. Therefore, in this experiment, we tried to develop ginsenoside Rb1 acupoint patch agent, use the acupoint patch agent to replace the acupuncture treatment, so that patients can easily accept the treatment, finally achieve the same therapeutic effect of acupuncture.

Babu agent is a form of drug taking hydrophilic polymer material as matrix mixed with different prescriptions to give play to the clinical therapeutic effect. Its affinity with the skin is good, it is non-allergic, drug loading is high, especially suitable for external prescriptions of traditional Chinese medicine with large dose and complex composition, and can satisfy the requirements of patients for comfortable pasting, convenient, low allergic and irritation, safe and effective use[10]. Therefore, it is feasible to use Babu agent as the drug carrier for invitro transdermal administration.

As the first step in the development of Babu agent, the prescription design of matrix is extremely crucial. Whether the matrix prescription design is scientific and reasonable will directly affect the performance of Babu agent. The biggest barrier to Babu agent is whether the drug can permeate the skin[11]. Therefore, it is necessary to study the invitro transdermal performance using Franz diffusion device. The results showed that Babu agent can make ginsenoside Rb1 permeate through the skin. With the extension of transdermal time, the cumulative permeation rate of ginsenoside Rbl also increases.

According to research findings, after the subjects pasting the ginsenoside Rb1 Babu agent, the amplitude of microvascular vasomotion was significantly increased (P<0.01). With the same effect after acupuncture, the increase in amplitude of microvascular vasomotion will generate beneficial effect on the body, it is helpful for blood flow, and suitable motion can promote cells to release various factors, favorable for cell metabolism and maintenance of cell structure, and such motion can be transmitted through cells and interstitial substance, and regulate the fusion reaction of particles in tissue fluid and exchange of substances between cells[12]. However, in the control group, after pasting the Babu patch, the amplitude of microvascular vasomotion of some subjects may also show significant increase (P<0.01), and the amplitude of increase was not as significant as pasting the Babu patch containing drugs. This is mainly because the transdermal enhancer has certain effect on the amplitude of microvascular vasomotion, but the effect is not as significant as the ginsenoside Rb1. The effects of drug-containing Babu agent on the amplitude of microvascular vasomotion is significantly different in different persons, but due to individual difference, the degree of influence may be different.

Using the people to carry out the experiment is mainly because people have control and can be free of external interference, and can keep a calm and steady stance over a period of time, which is difficult to achieve for other animals. Anesthesia can stabilize animals, but it can inhibit the amplitude of microvascular vasomotion[13-15]. Components of acupoint Babu agent are not nonpoisonous, need small invitro dose, and are not toxic to human body. In the future, Rb1 Babu agent is mainly used for the treatment of primary hypertension. Through overall consideration, we selected human beings as the experimental subjects. The experimental results indicate that the drug delivery system of ginsenoside Rbl Babu agent can release the drug into the acupoint, increase the amplitude of microvascular vasomotion, and achieve the effect of acupuncture. Therefore, ginsenoside Rbl Babu agent can replace the acupuncture clinically to treat diseases.

References

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由此揭示了个体发展与全体发展的互相促进、互为因果的关系，即发展不仅仅限于满足全体人民的物质文化需要，同样更要满足广大普通个体的生理与心理需求，才能达到社会的和谐。同样，在这段对科学发展观的经典论述中也寻不到社会或国家的影子。