Lumbar intervertebral disc herniation (LIDH) is a common and frequently-occurring disease with clinical symptoms of low back pain and/or sciatica[1]. Clinical and experimental studies have shown that electroacupuncture (EA) has confirmed therapeutic effects on LIDH. LIDH patients showed improved or alleviated clinical symptoms, such as lumbar and leg pain, limited motion and sciatica, after EA treatment[2-4], but the underling mechanism is still controversial. The clinical symptoms, physical signs, degree of intervertebral disc herniation and the nerve root compression are not exactly the same in LIDH patients. Therefore, the theory of nervous mechanical compression cannot fully explain the mechanism of LIDH[5]. Serum immunoglobulin G(IgG) and immunoglobulin M (IgM) are important products of the immune response in the body. The occurrence or increase of IgG and IgM in LIDH patients may be the results of autoimmune reaction in the prominent intervertebral disc[6]. Immunoassay for LIDH patients showed increased positive rate of IgG,suggesting that the immune response might be an important cause of LIDH[7]. In this study, we focused on the autoimmune reaction of LIDH to observe the changes of thigmesthesia function, gait function,expression levels of serum IgG and IgM in experimental rabbits with LIDH after EA at Huantiao (GB 30) and Weizhong (BL 40), thus to explore the related immunological mechanisms of EA for LIDH .

变压器是可以变换交流电压、电流和阻抗的一种静止电气设备。它的工作原理如下图所示，主要是利用电磁感应的原理，从一个电路向另一个电路传递电能或传输信号的一种电器。具体说来，变压器是由磁芯和线圈组成的，而线圈有两个或两个以上的绕组。其中，连接电源的绕组称之为初级线圈，剩下的绕组称之为次级线圈。当初级电圈中有交流电流时，磁芯就会产生交变磁通，此时，次级线圈就会感应到电流，两者产生电磁联系。根据电磁感应，交变磁通通过两个绕组就能感应出相应的电流。再当变压器空载时，初级电压(U1)/次级电压(U2)=初级圈数(N1)/次级圈数(N2)，电压的变化就会出现。

1 Experimental Materials

1.1 Experimental animals

A total of 40 healthy adult New Zealand purebred rabbits were provided by the Experimental Animal Center of Hunan Provincial People's Hospital Clinical Research Center (general level), half male and half female with body weight of 1.5-2.0 kg. Each rabbit was housed separately in the second laboratory of Experimental Animal Center in Hunan Provincial People's Hospital Clinical Research Center. Rabbits were fed at the temperature of 20-25 ℃, and humidity of 50%-70%, with sufficient light and free access to standard diet and drinking water. The handling of animals during the experiments was in line with the requirements of the Guiding Opinions on the Treatment of Experimental Animals issued by the Ministry of Science and Technology of the People's Republic of China[8].

1.2 Materials and reagents

Hwato Brand acupuncture needles of 0.30 mm in diameter and 25 mm in length (Suzhou Medical Appliance Factory, China); G6805-2 mode EA instrument (Qingdao Hua Qing Instrument Factory,China); IgG and IgM ELISA kit (Wuhan Xinqidi Biological Technology Co., Ltd., China).

1.3 Statistical methods

All data were analyzed using SPSS 19.0 software for Windows. First of all, normality test and homogeneity of variance test were performed. The data in normal distribution were presented as mean ± standard deviation (x±s). Paired t-test was used for intra-group comparison before and after intervention. One-way ANOVA was used to compare the differences among multiple groups. The least significant difference (LSD)was used when the variance was homogeneous and Tamhane's T2 method was used when the variance was not homogeneous. P＜0.05 indicated a statistical significance.

2 Experimental Methods

2.1 Animal groups

Using the quadratic random method, the rabbits were divided into a blank control group, a model group,an EA at acupoint group and an EA at non-acupoint group according to random number table method, with 10 rabbits in each group. All rabbits were fasted for 1 day, while with free access to water. Except for rabbits in the blank control group, rabbits in the other three groups were subjected to LIDH modeling[9]. Lumbar intervertebral disc CT examination was performed for the randomly selected rabbit samples after the successful modeling. Intervention started according to the grouping from the second day after modeling.

地理定位信息系统的主要原理是将与地理相关的内容进行电子化的转换，使其以动态的图形的形式呈现出来。这种信息系统在旅游管理中的应用主要体现在对游客所在的位置进行实时监控[1]。旅游管理人员通过这样的方式掌握游客的位置信息，可以为游客提供安全保障。所以，在旅游管理中应用地理定位信息系统，不仅能使游客感受到更加优质和完善的服务，还能为促进旅游管理信息化建设提供推动力。

2.2 Modeling method and standards for identification of the successful modeling

2.2.1 Modeling method

A LIDH model maker, reformed from a maxillary sinus puncture needle of 1.6 mm in diameter according to the literature, was used to establish the LIDH models[9].The median incision next to the left lower abdomen was performed under aseptic conditions to expose both the anterior longitudinal collateral ligament before the intervertebral space and the area behind the intervertebral space.

The LIDH model maker was inserted by 7.80-8.00 mm to the right with a 45° angle at the L6-L7 intervertebral space. Syringe was fixed and the intervertebral disc tissue in the syringe was push to the front of the posterior longitudinal ligament with a caput planum wick-in-needle, resulting in the right side LIDH model at L6-L7. Stitched the wounds, and the rabbits were sent back to the animal feeding room.

2.2.2 Standards to determine the success of models

据了解，《安徽省淮河流域综合治理规划纲要》期限为2011—2020年，重大项目展望至2030年及以后，规划区涉及10个省辖市的52个县区，2010年人口规模为4035万人，占全省的59%。规划提出，要以防洪保安、供水保障、生态保护为重点，加快推进淮河流域防洪、防旱、防污、供水综合治理，到2020年，基本建成与城镇化、工业化、农业现代化“三化同步”相适应的综合水利工程和水资源保障体系。

H社区志愿者消防队主要由社区成员、社区保安以及一些社区居民组成。 平时发生小型火灾，微型消防站第一时间需要到位，然后该社区物业管理公司的保安以及志愿者消防队的成员也会及时跟进，努力做到小型火灾自己扑灭，大型火灾尽量控制。 同时，通过调查了解，F市有几家消防器材公司会不定期地进入社区进行消防安全知识宣讲，但是主要以宣传公司产品为主，居民参与度较低。

Observation of lower limb nervous function: The lower limb nervous function of rabbits was observed and compared before and after modeling. Rabbit gait was scored according to the recommended neurological function criteria in the literature[10].

There were no statistically significant differences in neurological function scores of the lower extremities among groups before modeling. Except for the blank control group, the neurological function scores of rabbits’ lower extremity in the other three groups after modeling were significantly lower than those before modeling (P＜0.01), indicating that the gait function in rabbits was weakened after successful modeling.

CT examination: Lumbar CT examination for modeling rabbits was performed after anesthesia before and after modeling, respectively (Figure 1).

Rabbit's nucleus pulposus herniation to the internal spinal canal after modeling indicated the intervertebral disc herniation and successful modeling.

2.3 Point positioning

总之，语言中的文化负载词具有两大特点：(1)文化独特性。文化负载词只存在于特定的社会和生态环境中，没有特定的文化背景，就失去了存在的意义和价值。(2)等价性的困难。在两种不同的文化中，译者很难或几乎不可能找到两个完全等同的文化负载词。中华民族有上下五千年的漫长历史和灿烂的文化。在漫长的历史长河中，无数具有民族特色的词语被创造出来，在任何一个国家或民族中都找不到。只有不断深入地挖掘这些中国文化负载词，才能最大限度地消除中西文化的差异和冲突，实现顺利的跨文化交际。

Huantiao (GB 30): At the 1/3 middle and outside intersection of the line between the highest point of the femur greater trochanter and the sacral fissure of rabbit.

EA at non-acupoint group: Control points of Huantiao(GB 30) and Weizhong (BL 40) were selected. Negative pole was connected to the control point of Huantiao(GB 30), and positive pole was connected to the control point of Weizhong (BL 40). Acupuncture methods and EA parameters were the same as those in the EA at acupoint group.

Figure 1. CT findings before and after modeling

Control point of Huantiao (GB 30): Non-acupoint,2 cm above Huantiao (GB 30).

EA at acupoint group: Rabbits were fixed using the rabbit fixing box. Huantiao (GB 30) and Weizhong (BL 40)on the affected side were selected. After shearing and sterilizing, acupuncture needles of 0.3 mm in diameter and 25 mm in length were inserted directly by a depth of 1 cm. A group of output wires of the G6805-2 EA instrument were respectively connected to Huantiao(GB 30) and Weizhong (BL 40). Negative pole was connected to Huantiao (GB 30), and positive pole was connected to Weizhong (BL 40). Sparse-dense wave was used. The frequency of sparse wave was 30 Hz and the dense wave was 100 Hz. The current of 0.5-1.0 mA was used to keep the hind limbs of rabbits slightly tremulous.Each stimulus lasted for 20 min.

Control point of Weizhong (BL 40): Non-acupoint,0.5 cm medial to Weizhong (BL 40).

2.4 Treatment methods for rabbits in each group

Blank control group: Rabbits were fixed for 20 min using the rabbit fixing box without acupuncture treatment.

Model group: Rabbits were fixed for 20 min using the rabbit fixing box without acupuncture treatment.

Weizhong (BL 40): In the depression of the posterior knee joint.

（1）加强文化知识培训：文化程度对施工人员的影响是巨大的，尤其是对其自身施工的安全性将带来巨大的影响，主要体现在无法精准的掌握先进的施工工具，对安全管理认知度较低。因此本工程通过加强文化知识的培训，使其可以参加安全生产知识培训，促进安全意识的形成和完善。

2.5 Specimen collection and treatment

2.6.1 Thigmesthesia function test for the limbs

Point positioning was performed according to the commonly-used animal point positioning method in the Experimental Acupuncture Science[11].

2.6 Observed items

All rabbits were anesthetized with 20% urethane[4 mL/(kg·bw)]. Four milliliter of carotid artery blood was collected and kept at room temperature for 2-3 h,then centrifuged at 3 000 r/min for 15 min at 4 ℃.The supernatant was stored at -20 ℃ after aliquoted in EP tubes for later measurement.

Thigmesthesia function score was conducted by gently touching the affected toes with a cotton swab,according to the neurological function standards in the literatures[10]. 0 point: no any reaction; 1 point: slight reaction of affected limbs; 2 points: affected limbs had flexion or extension reaction, but the reaction was slow.3 points: reaction of affected limbs was quicker, but slightly worse than the healthy side; 4 points:thigmesthesia of the affected side was sensitive and showed no difference compared with the healthy side.2.6.2 Determination of limb gait function

The gait of rabbit was observed for gait score with reference to the literature[10]. 0 point: affected limbs were panplegia without autonomic activities; 1 point:affected limbs were semiplegia with muscle contraction and slight joint movement; 2 points: affected limbs showed poor strength, slow joint movement and walking instability; 3 points: affected limbs only showed interphalangeal joint dyskinesia; 4 points: recovered to normal exercise.

2.6.3 Serum IgG and IgM tests

Serum IgG and IgM were detected by enzyme-linked immunosorbent assay according to the kit instructions.

体育产业的崛起，为体育特色小镇发展提供了重要的资金支持，也为体育特色小镇发展提供了重要的人员循环参与的动力保障。当前，国家对体育事业的大力支持是体育产业发展的黄金时期，也是体育特色小镇建设的绝佳机遇。

3 Results

3.1 Results of thigmesthesia function

固定资产投资减去折旧等于当年的固定资产增量。如果假定技术不发生变化，用非工业部门当年的增加值相对于上一年的增量除以固定资产增量，就等于非工业产业单位增加值所需的直接固定资产含量。

3.2 Results of gait scores

There was no significant difference in gait score among groups before modeling (P＞0.05). The gait scores before and after modeling in the same group were compared: except for the blank control group, the gait scores of rabbits in the other three groups after modeling were significantly lower than those before modeling (all P＜0.01), indicating that the walking function was weakened after modeling. Comparison of the gait scores before and after treatment in the same group: the gait score after treatment in EA at acupoint group was significantly higher than that before treatment (P＜0.01), but no statistically significant intra-group differences in gait scores were found in other groups (all P＞0.05). Comparison of gait score difference after treatment among groups: EA at acupoint group was significantly higher than the blank control group, model group and EA at non-acupoint group (all P＜0.01), suggesting that the walking function of LIDH rabbits was significantly improved by EA at Huantiao (GB 30) and Weizhong (BL 40), thus showing a positive therapeutic effect on dysfunction of walking function in LIDH rabbits (Table 2).

Table 1. Comparison of rabbits’ thigmesthesia function scores (x±s, point)

Note: Compared with the score before modeling in the same group, 1) P＜0.01; compared with the score after modeling in the same group, 2)P＜0.01; compared with the blank control group, 3) P＜0.01; compared with the model group, 4) P＜0.01; compared with the EA at non-acupoint group, 5) P＜0.01

Group n Before modeling After modeling After treatment Difference before and after treatment Blank control 10 3.89±0.31 3.61±0.11 3.98±0.04 0.37±0.09 Model 10 3.87±0.32 1.80±0.421) 2.11±0.47 0.31±0.11 EA at acupoint 10 3.87±0.32 1.75±0.351) 3.63±0.462) 1.88±0.203)4)5)EA at non-acupoint 10 3.90±0.31 1.76±0.421) 2.14±0.47 0.38±0.11

Table 2. Comparison of rabbits’ gait scores (x±s, point)

Note: Compared with the score before modeling in the same group, 1) P＜0.01; compared with the score after modeling in the same group, 2)P＜0.01; compared with the blank control group, 3) P＜0.01; compared with model group, 4) P＜0.01; compared with the EA at non-acupoint group, 5) P＜0.01

Group n Before modeling After modeling After treatment Difference before and after treatment Blank control 10 3.92±0.16 3.61±0.11 3.98±0.04 0.37±0.09 Model 10 3.90±0.18 1.95±0.441) 2.24±0.40 0.29±0.07 EA at acupoint 10 3.93±0.16 1.87±0.321) 3.81±0.382) 1.94±0.183)4)5)EA at non-acupoint 10 3.93±0.16 1.91±0.201) 2.30±0.21 0.39±0.10

3.3 Serum IgG and IgM levels

Serum IgG and IgM levels in the model group were significantly higher than those in the blank control group (all P＜0.01). The serum levels of IgG and IgM in the EA at acupoint group were significantly lower than those in the model group and EA at non-acupoint group(P＜0.05). Serum IgG and IgM levels in EA at non-acupoint group were not statistically different compared with those in the model group, suggesting that the EA at Huantiao (GB 30) and Weizhong (BL 40)could inhibit serum IgG and IgM expressions in LIDH rabbits (Table 3).

There was no significant difference in thigmesthesia score among groups before modeling (P＞0.05). The thigmesthesia function scores of the same group before and after modeling were compared: except for the blank control group, the thigmesthesia scores of the other three groups after modeling were significantly lower than those before modeling (all P＜0.01),indicating that the thigmesthesia function of the rabbits after modeling was weakened. Comparing of the thigmesthesia function scores before and after treatment in the same group: thigmesthesia function score was significantly improved in the EA at acupoint group after treatment (P＜0.01), while there was no statistically significant difference in other groups before and after treatment (all P＞0.05). Comparison of thigmesthesia score differences after treatment among groups: the EA at acupoint group was significantly higher than the blank control group, model group and EA at non-acupoint group (all P＜0.01), suggesting that the thigmesthesia function was significantly improved by EA at Huantiao (GB 30) and Weizhong (BL 40), thus showing a positive therapeutic effect of EA at Huantiao(GB 30) and Weizhong (BL 40) on dysfunction of thigmesthesia function in LIDH rabbits (Table 1).

Table 3. Comparison of the levels of IgG and IgM expressions(x±s, μg/mL)

Note: Compared with the blank control group, 1) P＜0.01;compared with the model group, 2) P＜0.01; compared with the EA at non-acupoint group, 3) P＜0.01

Group nIgG IgM Blank control 10 8.04±1.15 6.72±1.39 Model 10 12.37±3.851) 10.33±3.141)EA at acupoint 10 9.01±2.832)3) 7.44±1.762)3)EA at non-acupoint 10 11.89±2.79 9.98±2.87

4 Discussion

LIDH falls under the category of ‘Bi-impediment syndrome’, ‘low back and leg pain’ or ‘Wei-flaccidity syndrome’ in Chinese medicine. It can be caused by either ‘obstruction’ or ‘malnourishment’. Obstruction occurs when exogenous wind, cold and dampness or traumatic injuries causes qi stagnation or blood stasis.Malnourishment occurs when long-lasting pain damages the liver and kidney or the failure of kidney qi to nourish the muscles or sinews due to constitutional liver and kidney deficiency. Consequently, the onset,development and treatment of LIDH are closely associated with meridians, qi, blood and functions of the Zang-fu organs.

Studies have found that the degree of lumbar disc herniation and nerve root compression, severities of clinical symptoms and physical signs in LIDH patients are not exactly the same[5]. Therefore, some people have questioned the opinion that compression and stimulationto the nerve root by the herniated nucleus pulposus are the main cause of low back and leg pain in LIDH patients[12]. It is believed that the mechanism of LIDH is not completely explained by mechanical compression of the nerve root alone, which may be closely related to the inflammatory response caused by biochemical substances in diseased intervertebral disc tissues[13] and autoimmune reactions[14-15]. Autoimmune response is also an important factor that causes low back pain and sciatica[14]. Intervertebral disc is an avascular tissue in the body. The nucleus pulposus is surrounded by the fibrous ring to make it isolated from the outside, thus has certain auto-immunogenicity, so the nucleus pulposus can be called ‘hidden antigen’. If the fibrous ring is ruptured, the nucleus pulposus will bulge from the ruptured fibrous ring and expose to the body's immune system, which can cause autoimmune reactions, lead to low back and leg pain, and other symptoms[14,16]. A study found that during the early phase of nucleus pulposus herniation in the noncompressive LIDH rats, T cell-mediated immune response caused nerve root injury, leading to radicular pain[17]. After LIDH modeling, IgG level in the body of rats was increased. Scraping treatment inhibits the autoimmune response induced by the nucleus pulposus and the inflammatory response mediated by autoimmune response, therefore, the immune abnormalities recover to the normal[18]. IgG and IgM are immunoglobulins in the body that reflect the body's immune status. The severity of LIDH is closely related to the expression level of IgG and IgM in the body[19-22].IgM shows stronger antigen binding ability and is the earliest appeared antibody during the initial humoral immune response. IgG is synthesized and secreted by the plasma cells in spleen and lymph nodes, and is the main antibody produced during the secondary humoral immune response. It is a high-affinity antibody that plays an important role in immune defense. Serum IgG and IgM are significantly elevated in LIDH patients, and acupuncture combined with herb-partitioned moxibustion can modulate humoral immunity in LIDH patients, leading to the IgG and IgM levels in patients to the normal[23]. Increased IgG and IgM levels in LIDH patients are thought to be the result of autoimmune response in intervertebral disc tissue[18]. Acupuncture can effectively reduce the levels of blood IgG and IgM in LIDH patients, decrease the secretion of immunoglobulin, and reduce the deposition of immune complexes in the lumbar intervertebral discs and surrounding tissues, thereby reducing the local inflammatory response in the intervertebral discs[24].

Area of the low back and leg pain caused by LIDH is similar to paths of the Bladder Meridian and Gallbladder Meridian. The two branches from the lumbus and back of the Bladder Meridian both meet at Weizhong (BL 40). Weizhong (BL 40) is the He-Sea point of the Bladder Meridian, and the common acupoint used in the treatment of low back and leg pain.Huantiao (GB 30) is a crossing acupoint of the Bladder Meridian and Gallbladder Meridian, and can be used to treat low back and leg pain related to the Bladder Meridian and Gallbladder Meridian. Gallbladder controls bone-induced diseases, and bladder controls tendon-induced diseases. Acupuncture at Huantiao(GB 30) and Weizhong (BL 40) has the role to smooth the tendons and help the joints, used in the treatment of low back and leg pain, bones and muscles pain of lower limbs and other related diseases.

国外的循证医学研究表明，抗病毒成分“金刚烷胺”对感冒病毒基本无效，感冒病毒对它耐药严重，因此国外不再把这个成分添加到复方感冒药中。中国药监部门已要求含“金刚烷胺”的感冒药修改说明书，对于可用于儿童、也可用于成人的氨酚烷胺胶囊，将“5岁以下儿童应在医师指导下使用”修订为“5岁以下儿童不推荐使用”，在【禁忌】项中增加“因缺乏新生儿和1岁以下婴儿安全性和有效性的数据，新生儿和1岁以下婴儿禁用本品”。

There are a lot of records about the treatment of low back and leg pain by Weizhong (BL 40) and Huantiao(GB 30) in the TCM classics from different ages.Therefore, Huantiao (GB 30) and Weizhong (BL 40) are the commonly used acupoints in the treatment of low back and leg pain. In this study, we observed the effects of EA at Huantiao (GB 30) and Weizhong (BL 40) on the expression of serum IgG and IgM in LIDH rabbits. We found that the thigmesthesia and walking functions of rabbits were decreased, and the levels of IgG and IgM in serum were increased significantly after modeling.After EA at Huantiao (GB 30) and Weizhong (BL 40),thigmesthesia and walking functions of rabbits were improved significantly, and serum IgG and IgM levels were significantly lower than those in the model group and EA at non-acupoint group; thigmesthesia and walking functions, and serum IgG and IgM levels in EA at non-acupoint group had no significant difference compared with those in the model group, suggesting that the autoimmune reaction may be related to the condition of LIDH, which is consistent with that being reported in domestic and foreign literatures[16,18,24],indicating that EA at Huantiao (GB 30) and Weizhong(BL 40) in regulation of LIDH rabbit serum immune response has a relative acupoint specificity effect. Based on the results above, it is concluded that the improvement of clinical symptoms, including thigmesthesia and gait score, of rabbits after treatment indicates that EA at Huantiao (GB 30) and Weizhong(BL 40) benifits LIDH rabbits. Prominent nucleus pulposus in LIDH rabbits can cause autoimmune response. EA at Weizhong (BL 40) and Huantiao (GB 30)can regulate the abnormal autoimmune reactions caused by autoantigen exposure due to the prominent nucleus pulposus in LIDH rabbits. Inhibition of the excessive immune response to nucleus pulposus autoantigen, thereby inhibiting of the initiation and progression of various inflammatory responses and alleviating clinical symptoms, may be one of the mechanisms during the treatment of LIDH. However,due to the small sample size, the limited experimental observation time (two courses) to detect the LIDH rabbit serum IgG and IgM levels in this trial, the real effect of EA at Huantiao (GB 30) and Weizhong (BL 40)on serum IgG and IgM levels in LIDH rabbit with a large sample size or under different experimental observation times remain to be further explored.

Conflict of Interest

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

最后，小微企业的发展离不开充足的资金支持，银行等金融机构要转变经营理念，结合当下大众创业、万众创新的时代潮流，为小微企业提供更为便捷的金融服务，解决小微企业发展的资金限制，优势互补，合作共赢。

This work was supported by Scientific Research Project of Hunan Provincial Administration of Traditional Chinese Medicine (湖南省中医药管理局科研资助项目, No.201378).

Statement of Human and Animal Rights

3.3 悬挂式土壤改良机工作原理 土壤改良机的运动流程见图3。由22.05 kW(30PS)拖拉机(张家口地区应用较广)提供动力，动力由液压油泵传输至液压马达和液压缸。液压马达带动钻头旋转，同时液压缸驱动升降架及钻坑部件垂直下行20～25 cm以形成深坑。螺旋钻罩的出土口定向排出钻土，坑深由位置传感器控制。深坑形成后液压缸驱动钻坑部件回程，待螺旋钻回到指定位置时，排肥机构中的步进电机精确转动实现定量出肥，肥料依靠重力和弯型排肥管独特的结构精准落入深坑，土壤改良机移动至下一工位的过程中覆土板带动少量薄土实现覆盖，完成深坑定量施肥。

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

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