[Effect of electroacupuncture on the morphological changes of the spinal dorsal horn and the expression of p38 mitogen-activated protein kinase in the injured spinal cord of rats with neuropathic pain].

OBJECTIVE: To investigate the effect of electroacupuncture stimulation at "Zusanli"(ST36) and "Kunlun"(BL60) on the morphological changes of the spinal dorsal horn and the expression of p38 mitogen-activated protein kinase (p38MAPK) in the injured spinal cord of rats with neuropathic pain. METHODS: Male Sprague-Dawley rats were randomly divided into sham model group, model group, electroacupuncture group, and medication group, with 10 rats in each group. Spinal nerve ligation of the L5 lumbar vertebra was performed to establish a rat model of neuropathic pain. The rats in the electroacupuncture group were given electroacupuncture at ST36 and BL60 of the operation side with dilatational wave at a frequency of 2 Hz/100 Hz and an intensity of 1.5 mA once a day, 30 minutes each time, and those in the medication group were given intraperitoneal injection of 100 mg/mL Gabapentin solution (100 mg/kg) once a day; the one-week intervention was started at one week after surgery. Mechanical withdraw threshold (MWT) and thermal withdrawal latency (TWL) were observed and recorded before modeling and on days 1,3,5,7,10,12 and 14 after modeling, and the motor function of the affected hindlimb was scored. Methenamine silver stain was used to observe the morphological changes of the spinal dorsal horn, and Western blot was used to measure the relative protein expression of p38MAPK and phospho-p38MAPK(p-p38MAPK) in L4-L6 spinal segments. RESULTS: Compared with the sham model group, the model group had significant reductions in MWT and TWL at each time point (P<0.001) and a significant increase in motor function score (P<0.001); compared with the model group, the electroacupuncture group and the medication group had significant increases in MWT and TWL and a significant reduction in motor function score after treatment (P<0.05). The model group had obvious neuronal fibrillary tangles, particle vacuolar degeneration, and vacuoles containing argyrophilic grains in the cytoplasm of neural cells under a light microscope, while there were fewer neuronal fibrillary tangles in the electroacupuncture group and reduced vacuolar degeneration in the medication group. Compared with the sham model group, the model group had significant increases in the protein expression of p-p38MAPK in the spinal dorsal horn (P<0.001), and compared with the model group, the electroacupuncture group and the medication group had significant reductions in the protein expression of p-p38MAPK in the spinal dorsal horn(P<0.05). CONCLUSION: Electroacupuncture stimulation at ST36 and BL60 can increase pain threshold, improve the motor function of the affected hindlimb, and improve the necrosis of neurofibrils in the spinal dorsal horn in rats with neuropathic pain, possibly by regulating the expression of p-p38MAPK in the spinal dorsal horn.

Combination treatment with Gua Sha and Blood-letting causes attenuation of systemic inflammation, activated coagulation, tissue ischemia and injury during heatstroke in rats.

OBJECTIVE: Gua Sha and Blood-letting at the acupoints were Chinese traditional therapies for heatstroke. The purpose of present study was to assess the therapeutic effect of Gua Sha on the DU Meridian and Bladder Meridian combined with Blood-letting acupoints at Shixuan (EX-UE 11) and Weizhong (BL 40) on heatstroke. METHODS: Anesthetized rats, immediately after the onset of heatstroke, were divided into four major groups: Gua Sha group, Blood-letting group, Gua Sha combined with Blood-letting group and model group. They were exposed to ambient temperature of 43 °C to induce heatstroke. Another group of rats were exposed to room temperature (26 °C) and used as normal control group. Their survival times were measured. In addition, their physiological and biochemical parameters were continuously monitored. RESULTS: When rats underwent heatstroke, their survival time values were found to be 21-25 min. Treatment of Gua Sha combined with Bloodletting greatly improved the survival time (230±22 min) during heatstroke. All heatstoke animals displayed and activated coagulation evidenced by increased prothrombin time (PT), activated partial thromboplastin time (aPTT), D-dimer, and decreased platelet count, protein C. Furthermore, the animals displayed systemic inflammation evidenced by increased the serum levels of cytokines interleukin-1ß (IL-1ß), tumor necrosis factor α (TNF-α) and malondialdehyde (MDA). Biochemical markers evidenced by cellular ischemia and injury/dysfunction included increased plasma levels of blood urea nitrogen (BUN), creatinine, serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), and alkaline phosphatase (ALP) were all elevated during heatstroke. Core temperatures (Tco) were also increased during heatstroke. In contrast, the values of mean arterial pressure were signifificantly lower during heatstroke. These heatstroke reactions were all signifificantly suppressed by treatment of Gua Sha and Blood-letting, especially the combination therapy. CONCLUSION: Gua Sha combined with Blood-letting after heatstroke may improve survival by ameliorating systemic inflflammation, hypercoagulable state, and tissue ischemia and injury in multiple organs.

[Anatomical characters and classification of acupoint].

OBJECTIVE: To explore the essence of acupoints by studying its anatomical characteristics and classification. METHODS: Based on numerous previous anatomic researches of acupoints, an adult male and a female corpses infused with red rubber at the location of acupoints were selected in order to observe the level and sectional anatomy of the acupoints, especially the specificity of the known structural distribution of acupoints. RESULTS: (1) The distribution of acupoints has two primary elements. Firstly, it is neurovascular bundles or connective tissue containing abundant nerves and blood vessels, which has the function of qi and blood infusion. It is considered as the acupoint kernel. Secondly, it is the cleft or the tunnel that formed by bones, muscles or fascia, which is also held as the acupoint shell. Different conditions of the human body can influence the opening and closing of the shell, control as well as regulate the function of the kernel so as to regulate the qi and blood circulation. (2) Based on anatomical characteristics, acupoints can be classified into 3 types: the type of cleft, the type of tunnel, and the terminal type. Points of the cleft and tunnel types can be mainly found in the twelve regular meridians which are composed of the kernel and the shell. The points of terminal type are on the Conception Vessel, the Governor Vessel and auricular point, the superficial fascia is the confluence of cerebro-spinal nerve terminal ramus and its concomitant vessels, and it is the site where the qi and blood of meridians distribute to the body surface. CONCLUSION: Acupoints is the pore formed by bone, muscle or fascia where neurovascular bundles or connective tissue containing abundant nerves and blood vessels pass through and the position where the terminal branch of cranial and spinal nerves and their accompanying by blood vessels emerge and converge at the supiyeficial layer of central body axis. Acupoints are the nodes and terminal point where the zang-fu organs and meridians and qi and blood infused to body surface.

[Characteristics of anatomical structures and clinical significance of lumbar Jiaji (EX-B 2) points].

OBJECTIVE: To observe the anatomical structure of Jiaji (EX-B 2) points at the level of lower lumbar region so as to provide evidence for the insertion angle and depth. METHODS: Thirty spine samples of male adults were adopted, and perpendicular insertion of the needle was applied at 3 locations including 1 cun, 0.5 cun and 0.3 cun lateral to the lower border of the spinous process of the lumbar vertebra. The needles were fixed at the local region. Structures and the adjacent major blood vessels and nerves were observed during the anatomy. RESULTS: When the needle was inserted perpendicularly at the point 1 cun lateral to the lower border of the spinous process of the lumber vertebra with the insertion depth of (35.77 +/- 5.86) mm, the zygapophyseal joints, the adjacent osteo-fibrous canal and osteo-fibrous aperture were touched by the tip of the needle, and the medial ramus of dorsal primary ramus of spinal nerve and concomitant vessels were stimulated. Then, needles were inserted perpendicularly 0.5 cun and 0.3 cun lateral to the lower border of the spinous process of the lumber vertebra with the insertion depth of (32.89 +/- 4.79)mm for both. When needle was inserted 0.5 cun lateral, the medial ramus of dorsal primary ramus of spinal nerve and the concomitant vessels were touched by the tip of the needle at where they across the lamina periosteum and erector spinae. When needle was inserted 0.3 cun lateral, the body of the needle reached the terminal branches of the medial ramus of dorsal primary ramus of spinal nerve and the concomitant vessels through the deep paraspinal muscles and the thoracolumbar fascia. CONCLUSION: The medial ramus of dorsal primary ramus of lumbar spinal nerve and concomitant vessels distributed at the region 1 cun, 0.5 cun and 0.3 cun beside the lower border of each lumbar spinous process. Therefore, the location of Jiaji (EX-B 2) points can be considered in the region from 0.3 cun to 1 cun beside the lower border of each spinous process.

[Morphologic characteristics and clinical significance of Neiguan (PC 6)].

OBJECTIVE: To provide appropriate needling angle and depth for the acupuncture and acupoint injection at Neiguan (PC 6), and to avoid damaging nerves and vessels so as to produce its maximum effect. METHODS: Thirty adults' upper-limb samples were used to dissect and observe the referred hierarchical structure and adjoining crucially nerves and vessels in needling Neiguan (PC 6) according to the national standard Acupoint Location (GB 12346-90). RESULTS: In this punctuation region, there are three parts rich in connective tissues containing the nerves and blood vessels. The surface part is between the skin and the musculus flexor digitorum superficialis and it is the tissue which contains medial and lateral antebrachial cutaneous nerve and its nutrient artery. The middle part is between the musculus flexor digitorum superficialis and the flexor digitorum profundus muscle and contains the median nerve, its palmar branch of and artery. The deep part is between the pronator quadratus muscle and the interosseous membrane and contains the anterior interosseous nerve. When perpendicular needling, the depth of needling the body from skin to the superficial surface of the musculus flexor digitorum superficialis and to the superficial surface of the flexor digitorum profundus muscle is (6.68 +/- 0.64) mm and (12.37 +/- 0.87) mm respectively. The depth of needling the body from skin to the superficial surface of the pronator quadratus muscle and to the superficial surface of the anterior interosseous terminal branch of the nerves is (17.83 +/- 1.00) mm and (30.87 +/- 1.85) mm respectively, and the proportional cun is (2.20 +/- 0.14) cm. The ulnaris cord of median nerves are located at the radial of the needle. The deep layers could touch the anterior interosseous nerve ending. CONCLUSION: Perpendicularly needling Neiguan (PC 6) for 3 fen (6.68 mm), 5 fen (12.37 mm) and 1.4 cun (30.87 mm) will stimulate the nervus vascularis of the rich part of surface, middle and deep connective tissues respectively and produce the acupuncture effect. During the acupoint injection, perforating the needle perpendicularly at the middle point of the two tendons or deviating slightly to the direction of tendon of palmaris longus can avoid the damage of the median nerve cord.

[Effect of electroacupuncture of local plus distal acupoints in the same segments of spinal cord on spinal substance P expression in rats with chronic radicular pain].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of local plus distal acupoints on spinal substance P expression in chronic radicular pain (CRP) rats so as to investigate its underlying mechanism in the treatment of chronic radical pain. METHODS: Twenty-five Wistar rats were randomized into control, model, local acupoints [LA, bilateral "Jiaji" (EX-B 2)], distal acupoints [DA, bilateral "Yanglingquan" (GB 34)], and LA+ DA groups, with 5 cases in each. CRP model was duplicated by implanting a gel-silicon wrapping the nerve root of L4 under anesthesia. EA (2 Hz, < or =2 mA) was applied to EX-B2 and GB34 for 30 min for 8 days. Pathological changes of the local focus tissue were observed by HE dyeing, and the animals' motor state was also observed. The pain threshold was detected by using tail-flick method. Substance P immunoreactive (IR) positive product of the spinal dorsal horn (L3-L5) was detected by immunohistochemical method and expressed as integrated optical density (IOD). RESULTS: The motor scores on day 35 after modeling and pain threshold values on day 4 and 8 after treatment in EX-B2 group, GB34 group and EX-B2 + GB34 group were significantly higher than those in model group (P < 0.05, 0.01). Compared with model group, IOD values of SP in the spinal dorsal horn in EX-B2 , GB34 and EX-B2 + GB34 groups were significantly lower (P < 0.05). There were no significant differences among the 3 EA groups in the expression of SP in the spinal dorsal horn. CONCLUSION: EA of EX-B2, GB34 and EX-B2 + GB34 all has a good analgesic effect in CRP rats, which may be realized partially by suppressing the release of SP in the spinal cord. No significant differences were found among local acupoint, distal acupoint and local plus distal acupoint groups in relieving CRP, improving motor and decreasing SP expression.

[Effect of Rhizoma curcumae oil on the learning and memory in rats exposed to chronic hypoxia and the possible mechanisms.].

The effect of Rhizoma curcumae oil on the learning and memory in rats exposed to chronic hypoxia and the possible mechanisms were investigated. The rats were divided randomly into 5 groups (14 animals in each group): control, chronic hypoxia, chronic hypoxia with low (5 mg/kg body weight), middle (10 mg/kg body weight) and high (20 mg/kg body weight) concentrations of Rhizoma curcumae oil injection. The animals undergoing chronic hypoxia were exposed to hypoxia in a hypoxic chamber containing 10% O(2) and 5% CO(2) for 10 h/d, lasting 28 d. Morris water maze (MWM) test was used to obtain the scores of leaning and memory. The superoxide dismutase (SOD) activity and malonaldehyde (MDA) content were determined in the serum and hippocampus as well as [Ca(2+)](i) in the hippocampus. The expression of phosphorylated Ca(2+)/calmodulin-dependent protein kinase II (p-CaMKII) in the hippocampus was evaluated by using immunohistochemistry and Western blot. Compared with the control group, the chronic hypoxia group showed the following changes: (1) The escape latency to the hidden platform was remarkably prolonged (P<0.05); (2) The content of MDA and [Ca(2+)](i) were obviously higher, but the activity of SOD and the expression of p-CaMKII were significantly lower (P<0.05, P<0.01). Compared with the chronic hypoxia group, groups with Rhizoma curcumae oil injection had the following changes: (1) The escape latency to the hidden platform was remarkably shorter in 10, 20 mg/kg body weight groups (P<0.05); (2) The content of MDA and [Ca(2+)](i) were markedly decreased in 5, 10, 20 mg/kg body weight groups (P<0.05, P<0.01), but the activity of SOD in the serum and the expression of p-CaMKII were significantly higher in 10, 20 mg/kg body weight groups (P<0.05, P<0.01). The results showed that the capacity of learning and memory was degraded following chronic hypoxia. The decrease in MDA content and [Ca(2+)](i) and (or) the increase in SOD activity and p-CaMKII expression might participate in the enhancing effect on learning and memory induced by Rhizoma curcumae oil.

[Research of integrated traditional Chinese and Western medicine on regularity of clinical acupuncture].

In the present article, the authors, in the light of their research in recent a decade of years, make an overall summary on the regularities of clinical acupuncture from the stimulated points, the superficial and deep layers, the literature basis and the innovation points. These basic regularities include symmetry or corresponding, segmental innervation, center or central axis, superiority of distal limbs, ending-stimulation accelerating analgesia, periphery-stimulation accelerating easiness and smooth, local feed back, specific feed back, selection of the stimulated points, acupuncture of different layers, etc.. They also discuss the possibility of integration of traditional Chinese and Western medicine from the view-points of neuroanatomy, neurophysiology and rehabilitation medicine. Systematic analysis on the regularities of clinical acupuncture makes the ancient meridian-collateral theory and abundant clinical experience summed up in definite concepts, simplifies the complicated theory system of acupuncture learning, and favors the communication about the terminology of Chinese medicine and Western medicine.

[Study on vascular morphology of integument tissues of the channel area in the leg].

OBJECTIVE: To observe the distribution of the blood vessels in the integument tissue of the channel area of legs. METHODS: The integument tissue of the lower limbs in the 12 cadavers were dissected with macro-and micro-dissection, radiographical technique of systemic artery and technique of image pattern analysis to observe and analyze the origins, branches and anastomoses in the integument tissues along the channels of legs. RESULTS: The distributional density of the blood vessels in the integument tissues of legs along the channel area of the three-yin meridians of the foot, the Gallbladder Meridian, and the Urinary Bladder Meridian was higher than that in the other parts. They formed an obvious nutrient vascular chain on the arteriogram. The distributional density in the channel area of the Stomach Meridian was not obviously increased and the obvious nutrient vascular chain could not be seen. CONCLUSION: An obvious nutrient vascular chain is formed in the integument tissue along the channel area of legs except the Stomach Meridian.

[Study on angle and depth of needle insertion in acupuncture at Zusanli (ST 36)].

OBJECTIVE: To provide an appropriate angle and depth of needle insertion in acupuncture at Zusanli (ST 36) and avoid injuring the nerve and blood vessel and exert the most effect. METHODS: Eighty adult lower-limb samples were used to dissect and observe the relative layered structures and adjoining important nerves and blood vessels in needling Zusanli (ST 36) according to the national standard. RESULTS: The needling depth from the skin to the interosseous membrane and from the skin to posterior border of tibialis posterior is (2.22 +/- 0.31) cm and (4.42 +/- 0.53) cm, respectively. There are flabellate branches of anterior tibial arteries and deep peroneal nerves around the needle in the superficial layer of interosseous membrane. The vessel and nerve bundles containing tibial nerve and posterior tibial vessels can be touched when the needle body past through tibialis posterior. CONCLUSION: It is recommended that ideal average depth of acupuncture is 2.22 cm and the maximum depth is 4.42 cm for oblique needling Zusanli (ST 36). When it is injected, the needle should be perpendicularly inserted or deviated slightly to the direction of tibia and paralleled to medial surface of tibia. And the safe needling depth is generally less than 5 cm. The point of the body surface between tibialis anterior and extensor digitorum longus at 3 cun below Dubi (ST 35) is also an effectively stimulating point.

[Anatomical study on Jingming (BL 1)].

OBJECTIVE: To explore the anatomical structures, and depth and direction of needling at Jingming (BL 1), so as to provide anatomical basis for its clinical application. METHODS: Forty-eight adult orbital specimens were observed by dissection. RESULTS: When a needle was vertically inserted into Jingming (BL 1), the needle tip will past through the skin, subcutaneous tissue, medial palpebral ligament, medialis rectus and orbital adipose body. Above the body of the needle, there are ophthalmic artery, anterior ethmoidal artery and nasociliary nerve. The average distance between the skin at the punctured point and the anterior ethmoidal artery is (18.25 +/- 4.45) mm, with an angle of (12.5 +/- 5.5) degrees, and the average distance between the skin at the punctured point and the optic nerve tunnel frontal point is (43.37 +/- 7.84) mm. CONCLUSION: To avoid bleeding caused by injuring the anterior ethmoidal artery, acupuncture at Jingming (BL 1) should avoid deeply inserting needled back-upwards and upwards, and the needling depth should not exceed 30.36 mm to avoid injury of the optic nerve tunnel frontal point.

Changes of learning, memory and levels of CaMKII, CaM mRNA, CREB mRNA in the hippocampus of chronic multiple-stressed rats.

BACKGROUND: The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not simulate complicated living circumstances that people confronted with. The aim of this study was to investigate the effects of chronic multiple-stress on learning and memory as well as on the levels of calcium/calmodulin-dependent protein kinase II (CaMKII), calmodulin (CaM) mRNA, and cAMP-response element binding protein (CREB) mRNA in the hippocampus of rats. METHODS: The rats were divided randomly into stressed and control groups. The stressed group was given chronic multiple-stress for 6 weeks to set up a chronic multiple-stressed model. The rats' performance of spatial learning and memory was tested using Morris Water Maze (MWM) and Y-maze. Meanwhile, the expressions of CaMKII, CaM mRNA and CREB mRNA of rats' hippocampus were detected by immunohistochemistry, Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. In addition, the width of synaptic cleft and the thickness of post-synaptic densities (PSD) were observed in the hippocampal CA3 region of rats by electron microscopy. RESULTS: After exposure to chronic multiple-stress for 6 weeks, the ability of learning and memory of the stressed group was higher than that of the control group (P < 0.05, P < 0.01). The width of synaptic cleft was smaller and the thickness of PSD was larger in the hippocampal CA3 region of the stressed group than in that of the control group (P < 0.01). The CaMK II immunostaining of the stressed group was stronger than that of the control group in the stratum radiatum and oriens of the hippocampal CA1 and CA3, especially in the stratum oriens. Quantitative analysis indicated that the expression of CaMK II, CaM mRNA, and CREB mRNA in the hippocampus of the stressed group was higher than that of the control group (P < 0.05, P < 0.01). CONCLUSIONS: The capacity of learning and memory can be enhanced after chronic multiple-stress. The increased levels of CaMK II, CaM mRNA, and CREB mRNA may contribute to the enhancing effect of chronic multiple-stress on learning and memory.

[Study on laws of clinical and basic integration acupuncture].

OBJECTIVE: To explore effective ways of integrated clinical and basic studies. METHODS: Try to sum up clinically basic laws of acupuncture from the study thinking of clinical and basic integration. RESULTS: Some laws of clinical acupuncture were raised based on classical theories of the channel and a great of clinical experiences, in combination with basic studies of modern neuroanatomy and physiology. CONCLUSION: The clinical symmetry-corresponding law, the segmental innervation law, the central-axial law, the distal limb dominance law, the law of sallow-deep correlation and the law of local feedback confirm with classical theories of the channels and clinical experiences.