Homing of the Stem Cells from the Acupoint ST-36 to the Site of a Spinal Cord Injury: A Preliminary Study

Homing of stem cells (SCs) to desired targets such as injured tissues remains a lingering problem in cell-based therapeutics. Studies on the biodistribution of intravenously administered SCs have shown the inefficacy of blood vessels as the homing path because most of the injected SCs are captured in the capillary beds of the lungs. We considered an alternative administration method using the acupuncture meridians or the primo vascular system. We injected SCs at the acupoint Zusanli (ST-36) below the knee of a nude mouse with a spinal cord injured at the thoracic T9-10 vertebrae. The SCs migrated from the ST-36, along the sciatic nerve, the lumbar 4-5, and then the spinal cord to the injury point T9-10. The SCs were not randomly scattered but were rather well aligned like marathon race runners, along the primo vascular system route toward the injury point. We observed the SCs at 1, 3, 6, 9, 12, and 15 hours after injection. The fast runners among the injected SCs took about 6 hours to reach the sciatic nerve, about 9 hours to reach the lumbar 4-5, and about 15 hours to reach the injury point T9-10.     

Primo-vessels and Primo-nodes in Rat Brain,Spine and Sciatic Nerve

We report a method using Trypan blue staining to detect primo-vessels in the nervous system on internal organs or in the skin of rat. We applied this technique to visualize the primo-vessels and primo-nodes in the brain, spinal cord and sciatic nerve of a rat. Primo-vessels and primo-nodes were preferentially stained at nerves, blood vessels, or fascia-like membranes and turned blue after the spread and washing of Trypan blue. The physiological role of the primo-vessels within the nervous system is an important question warranting further investigation.     

Primo-Vessels as New Flow Paths for Intratesticular Injected Dye in Rats

After intratesticular injection of a chromium hematoxylin and fluorescent nanoparticle solution, we found a novel flow path in the abdominal cavity consisting of primo-vessels and primo-nodes. This flow path formed a network that crossed over the surfaces of abdominal organs, and generally linked to the greater omentum and adjacent visceral peritoneum. Some of these structures terminated at organs such as the small intestine and the urinary bladder; occasionally, the network entered the parenchyma of organs. The semitransparent primo-vessels and nodes were wholly or partially stained dark-blue by chromium hematoxylin. Injected nanoparticles were also observed in primo-vessels and nodes as well as the parenchyma of organs which were the site of primo-vessel termination. Transmission electron microscopy showed that the primo-vessels consist of many sinuses (4–6 μm), surrounded by collagen fibers, specific granules, cellular remnants, immune cells, extracellular matrices, and hematopoietic cells. These histological features are completely different from blood and lymph vessels indicating that primo-vessels are novel structures that allow the flow of dye.     

手针刺激“足三里”穴局部肥大细胞活动与外周神经放电的相关性研究

目的:探讨穴位处肥大细胞的激活及穴区胶原与支配该区域的神经束放电变化之间的关系。方法:雄性SD大鼠30只,随机分为空白组、对照组、手针足三里组、手针+色甘酸钠组(穴区肥大细胞拮抗)、手针+胶原酶组(穴区胶原纤维溶解),每组6只。动物麻醉后,分离左侧"足三里"穴区坐骨神经干,手针提插刺激大鼠"足三里"穴20min,记录刺激前、刺激时和刺激后神经束诱发放电信号。对穴位组织进行组织学观察,统计肥大细胞脱颗粒率。结果:与对照组、手针+色甘酸钠组和手针+胶原酶组相比,手针足三里组诱发的坐骨神经放电活动明显增强(P<0.01);同时手针足三里组肥大细胞脱颗粒率显著升高(P<0.01),而手针+色甘酸钠组和手针+胶原酶组肥大细胞脱颗粒率与对照组相比差异无统计学意义(P>0.05)。结论:手针刺激大鼠"足三里"穴位可诱发支配该区域的外周神经束放电;穴位区注射色甘酸钠可抑制肥大细胞脱颗粒,针刺引起的外周神经放电减弱;穴位区注射胶原酶阻断针刺应力在穴位组织中传输,可以抑制肥大细胞脱颗粒,同样,针刺引起的外周神经放电也减弱。     

粘合吻合法与神经导管修复神经的实验研究

目的探讨应用粘合吻合法及神经导管修复神经损伤的可行性,并评估其修复效果。方法SD大鼠50只,随机分为粘舍吻合组与缝合组,每组25只。以右侧坐骨神经建立修复模型。粘合组用α-氰基丙烯酸酯医用粘合剂与优化外形的神经导管修复神经断端,缝合组用9—0的显微缝合线做神经外膜缝合6针。分别记录手术操作时间。通过测量术后2,4,8,12周坐骨神经指数,以及电生理检测、腓肠肌湿质量恢复率和组织学观察评估修复效果。结果粘合纽操作时间较缝合组明显缩短,坐骨神经指数、电生理检测、腓肠肌湿质量及组织学观察2组间比较均无显著性差异。结论应用粘合吻合法与神经导管修复神经缺损保证修复效果的同时,大大缩短了手术时间,是一种简便、有效的神经修复方法,具有一定的临床应用前景。     

Study of novel threadlike structures on the intestinal fascia of dogs

The primo-vascular system was visualized in the mesentery surrounding the small intestine of a dog using Trypan blue. This structure, which was first observed in a rat, formed a network as primo-vessel branches were joined to primo-nodes. Other characteristic features of the primo-vascular system, such as bundles of tubes with fibrous extracellular matrix in a primo-vessel and a broken-line alignment of rod-shaped nuclei along the primo-vessel, were observed. Blood vessels, lymph vessels, and primo-vessels were present in the same mesentery, and they could clearly be distinguished by histological differences.     

Putative primo-vascular system in mesentery of rats

Primo-vessels have been observed in the rat abdominal cavity as floating thread like structures on and not adhering to fascia-wrapped internal organs. To date their presence, locations, and lengths have been irregular and unpredictable, and their identification not regularly repeatable, thus they have remained a nagging enigma in primo-vascular system research for several years. In this work, locations were found where primo-vessels were regularly present and observed repeatedly. These vessels were not floating or freely movable but lay in a regular position in the mesentery in the abdominal cavity of the rat, being observed between the cecum and small intestine and between the colon and mesentery root. The difference between a lymph vessel and a primo-vessel is described in anatomical and histological aspects. In addition, trypan blue was found to enter primo-vessels through the surrounding membranes and filled spaces between fibers comprising the primo-vessels. It is conjectured that the previously observed floating primo-vessels had anomalously and irregularly emerged, for some unknown physiological reasons, from primo-vessels normally located in the fascia-like mesentery.     

大鼠"承山"穴和坐骨神经之间的神经解剖学联系

Objective:The aim of this study is to assess the neural interconnection between the acupoint"Chéngshān(承山BL57)"and sciatic nerve from the peripheral nervous sys...     

Electroacupuncture improves neuronal function by stimulation of ascending peripheral nerve conduction in rats with spinal cord injury

OBJECTIVE: To establish a method of transient sciatic nerve blockade and to examine the involvement of the ascending peripheral nerve pathway in the therapeutic effect of electroacupuncture at Zusanli(ST 36) in rats with spinal cord injury(SCI).METHODS: We examined the transient effect of daily lidocaine administration into the posteromedial part of the greater trochanter on sciatic nerve function using electrophysiological examination and histopathology of the sciatic nerve. Rats were divided into three groups: an SCI group(SCI without treatment), an SCI with electroacupuncture treatment(SCI-EA) group, and an SCI with nerve block and electroacupuncture(SCI-NB-EA) group(nerve block was achieved by lidocaine administration to transiently block the ascending peripheral nerve pathway). Behavioral tests and electrophysiological examinations were performed to evaluate recovery of neurological function.RESULTS: Sciatic nerve conduction was normal immediately before daily lidocaine administration.Histopathological analysis also indicated normal sciatic nerve, confirming that lidocaine nerve blockade was suitable and reversible for transiently eliminating nerve transmission. Neurological function in the SCI-EA group was superior to that in the SCI group, while no differences were found between the SCI and SCI-NB-EA groups.CONCLUSION: Electroacupuncture treatment can promote recovery of neurological function. Facilitation of nerve conduction may play an important role in this recovery.     

Effects of acupuncture on potential along meridians of healthy subjects and patients with gastric disease

The purpose of this study was to investigate the differences in bio-potential in the stomach meridians of 20 normal healthy control subjects (NHC) and 20 patients with gastric disease (PGD). We measured changes in potential between ST-39 and ST-37 on the stomach meridian in response to invasive insertion of a needle at ST-36. The response patterns of NHC were regular in both the left and right meridians, whereas the response patterns of PGD were irregular. The potential of the NHC was significantly higher than that of the PGD group. The potential values in the left and right meridians of NHC were the same. However, the potential in the right meridian was significantly higher than that in the left meridian of PGD (p < 0.001). This study indicates that there may be changes in Qi or in the electrical properties of meridians according to health status, reflected in the regularity of the response to acupuncture or in the level of potential along meridians. Therefore, it may be possible to make a diagnosis from the state of a meridian by comparing levels of potential difference and its regularity.     

Neuroprotective effect of saponin rich extract of Acorus calamus L. in rat model of chronic constriction injury (CCI) of sciatic nerve-induced neuropathic pain

Ethnopharmacological relevance

Traditionally, Acorus calamus has been used for the treatment and management of headache, migraine, body ache and severe inflammatory pain in the Unani, Ayurveda and Indian system of medicine.

Aim of the study

Present study focuses on the evaluation of saponin rich extract of Acorus calamus (SRE-AC) in chronic constriction injury (CCI) of sciatic nerve induced neuropathic pain and neuronal functional changes in rats.

Materials and methods

The pain sensitive tests, i.e., thermal and mechanical hyperalgesia and sciatic functional index test, were performed on the different days, i.e., days 0, 1, 7, 14, and 21. The motor and sensory nerve conduction velocity was also measured on the 21st day. Tissue total protein, superoxide anion generation, total calcium, myeloperoxidase and TNF-α levels were estimated to assess biochemical changes. Histopathological evaluations were also performed. SRE-AC (20 and 40 mg/kg) and pregabalin (10 mg/kg, serving as a positive control) were administered orally for 14 consecutive days from the day of surgery.

Results

CCI produced significant (P<0.05) increase in thermal and mechanical hyperalgesia, rise in sciatic functional index, decrease in nerve conduction velocity, along with biochemical and histopathological changes. Oral administration of SRE-AC and pregabalin significantly (P<0.05) ameliorated CCI-induced nociceptive pain threshold, sciatic functional and electrophysiological changes in a dose dependent manner. Further, tissue biochemical and histopathological changes were also attenuated.

Conclusion

SRE-AC has shown ameliorative effect in CCI-induced neuropathic pain which may be attributed to its multiple actions including anti-oxidative, anti-inflammatory and neuroprotective actions.     

针刺结合改良神经外膜束膜联合缝合术治疗周围神经损伤的临床观察

目的观察针刺联合改良神经外膜束膜联合缝合术治疗周围神经损伤的临床疗效。方法将60例(72条神经)周围神经损伤患者,随机分为治疗组与对照组,每组30例。对照组行改良神经外膜束膜联合缝合术及常规康复治疗。治疗组在对照组治疗措施基础上,加用针刺治疗。两组疗程均为6个月,观察神经功能疗效及电生理疗效,比较神经电生理指标(传导速度与波幅)的变化情况。结果 1治疗组、对照组神经功能优良率分别为86.11%、69.44%;组间神经功能疗效比较,差异有统计学意义(P0.05)。2治疗组、对照组神经电生理总有效率分别为91.67%、75.00%;组间电生理疗效比较,差异有统计学意义(P0.05)。3治疗前后组内比较,两组神经传导速度、波幅差异均有统计学意义(P0.05);组间治疗后比较,神经传导速度、波幅差异有统计学意义,治疗组优于对照组(P0.05)。结论针刺结合改良神经外膜束膜联合缝合术治疗周围神经损伤的疗效满意,有助于患者神经功能的恢复。     

The hypoglycemic activity of Lithocarpus polystachyus Rehd. leaves in the experimental hyperglycemic rats

Ethnopharmacological relevance

Leaves of Lithocarpus polystachyus Rehd. are used for the treatment of disorders such as diabetes, hypertension, and epilepsy in folk medicine of South China. The possible antidiabetic effects of the leaves were investigated in experimental type 2 and type 1 diabetic rats.

Materials and methods

Type 2 diabetic rats received orally three different extracts of Lithocarpus polystachyus Rehd. leaves for 4 weeks (aqueous extract [ST-1], ethanol extract [ST-2], flavonoid-rich fraction [ST-3]). At the end of the experiment biochemical parameters were tested and livers and pancreases were excised for histological study. After the comparison of the pharmacological test results of the three extracts, the one which showed the best bioactivity was further studied to confirm its antidiabetes effect on both type 2 and type 1 diabetic rats.

Results

Compared to ST-1 and ST-2, ST-3 had better effects on regulation of blood glucose, glycosylated serum protein, cholesterol, triglyceride, malondialdehyde, superoxide dismutase and attenuation of liver injury in type 2 diabetic rats (p < 0.01 or p < 0.05). ST-3 administration for four weeks also significantly reduced the fasting serum insulin and C-peptide level and improved the insulin tolerance (p < 0.05). In type 1 diabetic rats, ST-3 supplement for three weeks caused significant reduction in fasting blood glucose, total cholesterol, triglyceride, urea nitrogen, creatinine and liver mass, along with significantly inhibiting the decline of insulin level compared to diabetic control (p < 0.05 or p < 0.01).

Conclusion

The flavonoid-rich fraction of Lithocarpus polystachyus Rehd. leaves (ST-3) had better beneficial effect than that of the ethanol or aqueous extract in experimental diabetic rats, which means that the bioactivity of the herbal leaves is probably due to the presence of flavonoids. The results also strongly suggest that the antidiabetic effect of ST-3 was possibly through multiple mechanisms of action including blood lipid and antioxidant mediation. The results indicated that the aqueous flavonoid-rich fraction of Lithocarpus polystachyus Rehd. leaves possessed significant protective activity in type 2 and type 1 diabetes.     

不同频率电针对大鼠坐骨神经损伤后神经组织形态学与骨骼肌肌电图的影响

目的:观察不同频率电针对再生过程中神经组织形态学以及骨骼肌肌电图(EMG)的影响,寻找较适宜的电针参数.方法:构建坐骨神经截断后神经再生室,以5 Hz和100 Hz电针刺激患侧"环跳""足三里"和"三阴交",每次治疗10 min,隔日1次,每周治疗3次;并设对照组观察.各组治疗时间为20 w.以神经纤维银染以及超微结构观察电针治疗组和对照组的再生组织形态学变化;应用Biopac多导电生理仪,测定腓肠肌EMG.结果:电针刺激能明显促进坐骨神经损伤后神经组织形态及失神经支配腓肠肌功能的恢复,其中以5 Hz电针组治疗效果最佳.结论:电针是促进周围神经损伤再生的重要手段.     

穴位注射与针刺拔罐结合治疗坐骨神经痛

坐骨神经痛是指沿坐骨神经通络及其分布区的疼痛,分原发性和继发性两类。前者是由感染受寒,中毒等原因直接损害坐骨神经所引起的,后者是有小中经通络近组织的病变,对坐骨神经产生刺激,压迫粘连或破坏而造成的,根据神经受损的部位又分为根性和干性。其中以干性多见。     

艾灸对原发性坐骨神经痛大鼠坐骨神经干及脊髓腹角GAP-43表达的影响及其神经修复作用

目的:观察艾灸"环跳"穴对原发性坐骨神经痛大鼠坐骨神经干和脊髓腹角(L₄~L₆)生长相关蛋白-43(GAP-43)表达的影响,探讨艾灸改善原发性坐骨神经痛的作用机制。方法:将48只健康雄性SD大鼠随机分为正常组、假手术组、模型组和艾灸组,每组12只。模型组和艾灸组大鼠采用坐骨神经慢性压迫性损伤(CCI)术建立原发性坐骨神经痛大鼠模型。实验第8天,艾灸组大鼠于患侧"环跳"穴行温和灸5~10 min,每日1次,连续14 d。分别于实验第1、7、14、21天测定各组大鼠坐骨神经功能指数(SFI)。实验第21天,HE染色法观察大鼠脊髓腹角和坐骨神经干形态;免疫组织化学法和实时荧光定量PCR法检测大鼠脊髓和坐骨神经干GAP-43蛋白和mRNA的表达。结果:假手术组大鼠实验第7、14、21天SFI与正常组比较差异无统计学意义(P>0.05);与假手术组比较,模型组大鼠实验第7、14、21天SFI降低(P<0.01);与模型组比较,艾灸组大鼠实验第14、21天SFI升高(P<0.01)。正常组和假手术组大鼠脊髓腹角内神经元细胞排列整齐,尼氏体分布均匀,坐骨神经轴突髓鞘结构清晰可见;模型组大鼠脊髓腹角内可见神经元细胞变形和破溃,尼氏体数量少,坐骨神经轴突脱髓鞘改变;艾灸组大鼠脊髓腹角内可见神经元细胞排列较规则,尼氏体增多,坐骨神经轴突部分脱髓鞘改变。与正常组比较,假手术组大鼠坐骨神经干GAP-43 mRNA和蛋白表达、脊髓腹角GAP-43蛋白表达升高(P<0.01);与假手术组比较,模型组大鼠脊髓腹角和坐骨神经干GAP-43 mRNA和蛋白表达升高(P<0.01);与模型组比较,艾灸组大鼠脊髓腹角和坐骨神经干中GAP-43 mRNA和GAP-43蛋白表达升高(P<0.01)。结论:艾灸"环跳"穴可改善原发性坐骨神经痛大鼠坐骨神经功能,可能与上调脊髓腹角和坐骨神经干GAP-43表达,增强坐骨神经的自我修复能力有关。     

手法治疗坐骨神经痛

坐骨神经是指坐骨神经病变,沿坐骨神经通路,即腰,臀部,大腿后,小腿后外侧合足外侧发生的疼痛症。它多发于男性青壮年,此病给患者的生活工作带来许多不便。     

Visualization of Acupuncture Meridians in the Hypodermis of Rat Using Trypan Blue

Bonghan Kim claimed that the primo-vessel (Bonghan duct) and the primo-node (Bonghan corpuscle) are anatomical structures corresponding to acupuncture meridians and acupoints, respectively, but this claim has not been confirmed. Recently we found that Trypan blue preferentially stained primo-vessels and primo-nodes on the surfaces of internal organs, not staining other structures like blood or lymph vessels or nerves. In this work, we applied Trypan blue to visualize the putative acupuncture meridian along skin skeletal muscles in the hypodermal layer of a rat. The Trypan blue stained-structures are morphologically similar to the meridian primo-vessels, as claimed by Bonghan Kim. Further study is needed to investigate the network of these Trypan blue-stained structures in order to establish them as acupuncture meridians.     

Regulatory Effect of Electroacupuncture on Ceramide Galactosyltransferase Expression in Sciatic Nerve of Experimental Diabetic Rats简

Objective： To observe the change of ceramide galactosyltransferase（CGT） expression in sciatic nerve of experimental diabetic rats after electroacupuncture（EA） treatment. Methods： The rat model of diabetic peripheral neuropathy（DPN） was established with Streptozotocin（STZ）. Twenty-eight rats were randomly divided into a normal group, a model group, an EA acupoint group, and an EA non-acupoint group, with 7 rats in each group. Acupuncture on points Shenshu（BL 23） and Zusanli（ST 36） was performed in the EA group using EA electronic device with continuous wave, 2 Hz, 20 min every time, once every other day for 12 times. Acupuncture（on the tip of rat tail） in EA non-acupoint group was performed with the same electronic parameters and time. The model group and the normal group were not given any treatment, except the same fixation as that in the EA acupoint group. After 12 treatments, the albumen and mRNA expressions of CGT of sciatic nerve in rats from each group were measured with real-time polymerase chain reaction（RT-PCR）, fluorescence quantitative PCR（FQ-PCR） and immunohistochemical methods. Results： After modeling, the CGT expression was significantly lower than that in the normal group（P〈0.01）; however, after 12 EA treatments, the CGT positive cells in the EA acupoint group were up-regulated and significantly higher than those in the EA non-acupoint group and the model group（P〈0.01）. The CGT mRNA expression of the rats after modeling was also higher than that of the normal rats（P〈0.01）, and the expression in the EA acupoint group was significantly lower than those in the model group and the EA non-acupoint group（P〈0.01, P〈0.05）. Conclusion： EA treatment with acupoints displays some therapeutic or preventive effect in the recovery of DPN via reversing the abnormal expression of CGT mRNA in sciatic nerves of experimental diabetic neuropathy rats, and it is worthy of further study.     

手针和电针大鼠“足三里”穴位诱发的外周传入神经信号特征研究

目的:观察手针、电针刺激大鼠"足三里"穴区时支配该区域的神经束诱发的放电变化,比较不同参数的手针和电针诱发的神经束放电的差异,初步揭示针刺信息的外周传导规律。方法:30只雄性SD大鼠,随机分为手针足三里组和电针足三里组,每组15只。动物用20%乌拉坦麻醉(750 mg/kg),分离支配右侧"足三里"穴区坐骨神经分支后,分别以不同手法的手针(捻转、提插、捻转提插,1 Hz)及不同参数的电针(5 V、10 V1、5 V,3 Hz)刺激大鼠"足三里"穴各1 min,对刺激过程及刺激前后神经束诱发放电记录采样各1 min,然后进行单位时间神经放电脉冲计数(Bin:1 s)。结果:捻转提插手针"足三里"其神经束放电峰电位数增加最为明显:刺激前的计数为(42.20±1.98)pulses/s;刺激后增为(74.20±18.92)pulses/s(P<0.05);刺激结束后即刻恢复到(41.40±4.85)pulses/s,与刺激后比较P<0.05。单纯提插手针刺激时其神经束放电峰值计数从刺激前的(42.00±3.89)pulses/s增加为(60.60±10.79)pulses/s(P<0.05);针刺结束后即刻恢复至(45.00±18.10)pulses/s,与刺激后比较P<0.05。而单纯捻转手针刺激无明显变化。电针刺激"足三里":强度从5 V左右开始可诱发类似手针诱发的神经放电现象,10 V以上电针其神经放电增加显著,15 V电针接近于单纯提插手法手针所产生的效应:放电峰电位数从刺激前(42.60±3.87)pul-ses/s增加至电针时(64.20±9.36)pulses/s(P<0.05);电针结束后恢复到(48.00±7.55)pulses/s,与电针时比较P<0.05。结论:提插及捻转提插手法的手针刺激大鼠"足三里"可诱发支配该区域的外周神经束放电,且后者放电现象更为明显;一定强度的电针刺激可产生与手针的机械刺激类似的神经放电效应。