[Effect of acupuncture at "Houxi" (SI3) and"Huantiao" (GB30) on high mobility group box 1 protein in spinal nerve trunk of rats with lumbar disc herniation].

OBJECTIVE: To observe the effect of acupuncture at "Houxi"(SI3) and "Huantiao"(GB30) on high mobility group box 1(HMGB1) protein and mRNA in spinal nerve trunk(SNT) of rats with lumbar disc herniation(LDH), so as to explore the mechanisms of acupuncture at this paired points on the treatment for LDH. METHODS: SD rats were randomly divided into sham operation, model, conventional acupuncture(CA) and paired points(PP) groups (with 8 rats in each group). The LDH model was established by injection of autologous suspension made from rats' own nucleus pulsus into the epidural space. Rats in the CA group received acupuncture treatment at bilateral "Weizhong"(BL40), "Dachangshu"(BL25) and "Shenshu"(BL23), while rats in the PP group received acupuncture at bilateral SI3 and GB30, 30 min each time, once daily for 14 consecutive days. The thermal pain threshold of bilateral hind feet of rats was detected by thermal pain stimulator. The contents of serum IL-1ß, IL-6 and IL-8 of rats were detected by ELISA. Western blot and immunofluorescence were used to detect the expression of HMGB1 protein in the lumbar(L)5 SNT of rats. The relative expression of HMGB1 mRNA in L5 SNT was determined by qPCR. HE staining was used to observe the morphological changes of L5 SNT. RESULTS: Compared with the sham operation group, the thermal pain threshold of bilateral hind feet in the model group was decreased (P<0.05); compared with the model group, the thermal pain threshold of bilateral hind feet in the CA group and the PP group were increased (P<0.05). The expressions of HMGB1 protein and mRNA in L5 SNT, and the contents of serum IL-1ß, IL-6 and IL-8 of rats in the model group were significantly increased(P<0.000 1, P<0.001) in contrast to the sham operation group. The expressions of HMGB1 protein and mRNA in L5 SNT, and the levels of serum IL-1ß, IL-6 and IL-8 were significantly decreased (P<0.01, P<0.000 1, P<0.001, P<0.05) in the CA and PP group, in comparison with those of the model group. Compared with the CA group, the above indexes of rats in the PP group recovered more significantly (P<0.05,P<0.001, P<0.01,P<0.000 1). The histomorphological results showed scattered and various-sized nerve fibers, vacuolation, a large number of disintegrating myelin sheath and lysed Schwann cells in the model group. Myelin sheaths regeneration, regularly-arranged nerve fibers were seen in the CA group and the PP group, with more obvious histopathological recovery observed in the PP group than the CA group. CONCLUSION: Acupuncture intervention inhibites the expressions of HMGB1 protein and mRNA in rats with LDH, and further reduces the production of IL-1ß, IL-6 and IL-8, which is beneficial to inflammatory response inhibition and pain alleviation. The therapeutic effect of the PP group is more obvious than that of the CA group.

[Effect of wheat-grain moxibustion on the expression of 5-HT and cortisol in the serum, and MR and GR in the hippocampus in rats with hypothyroidism complicated with depression].

OBJECTIVE: To observe the effect of wheat-grain moxibustion on behavior, 5-hydroxytryptamine (5-HT) and cortisol in the serum, mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) in the hippocampus in rats with hypothyroidism complicated with depression, and to explore the possible mechanism of wheat-grain moxibustion on improving depression in rats with hypothyroidism. METHODS: A total of 32 SPF SD rats were randomly divided into a blank group, a model group, a medication group and a wheat-grain moxibustion group, 8 rats in each group. Except for the blank group, the rats in the remaining groups were treated with intragastric administration of 0.1% propylthiouracil (PTU) suspension at 1 mL/100 g, once a day for 4 weeks to establish the rat model of hypothyroidism, and whether the rats were accompanied with depression-like behavior determined through behavioristics evaluation. The rats in the medication group were intervened with euthyrox at 0.9 mL/100 g, once a day, for 4 weeks; the rats in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14), "Mingmen" (GV 4), "Shenshu" (BL 23) and "Pishu" (BL 20), 7 cones each acupoint, once a day, six times a week for 4 weeks. After the intervention, the depression status was observed by behavioristics test; the contents of thyroid stimulating hormone (TSH), total thyroxine (TT4), 5-HT and cortisol in the serum were detected by ELISA; the protein expressions of MR and GR in hippocampus were detected by Western blot; the expressions of MR mRNA and GR mRNA in the hippocampus were detected by real-time PCR. RESULTS: Before the intervention, compared with the blank group, the scores of open field test (OFT) were decreased and the immobility time of tail suspension test (TST) was prolonged (P<0.05); the serum TSH contents were increased and TT4 contents were decreased (P<0.01) in the other three groups. After the intervention, compared with the model group, the vertical score of OFT was increased and the immobility time of forced swimming test (FST) was prolonged in the medication group (P<0.05), while the scores of three items of OFT were increased (P<0.05, P<0.01), and the immobility time of FST and TST was shortened in the wheat-grain moxibustion group (P<0.01, P<0.05). Compared with the medication group, the immobility time of TST and FST in the wheat-grain moxibustion group was shorter (P<0.05, P<0.01). Compared with the blank group, in the model group, the contents of serum TSH and cortisol were increased (P<0.01, P<0.001), while the contents of serum TT4 and 5-HT were decreased (P<0.01, P<0.001). Compared with the model group, the contents of serum TT4 and 5-HT were increased, while the contents of serum TSH and cortisol were decreased in the medication group and wheat-grain moxibustion group (P<0.01, P<0.05). Compared with the blank group, the protein and mRNA expression of MR, GR in the hippocampus in the model group was decreased (P<0.01, P<0.05, P<0.001); compared with the model group, the protein and mRNA expression of MR in the hippocampus in the medication group were increased (P<0.05), and the protein expression of MR, GR and mRNA expression of MR in the hippocampus in the wheat-grain moxibustion group were increased (P<0.05, P<0.01). Compared with the medication group, the expression of MR mRNA in the wheat-grain moxibustion group was increased (P<0.05). CONCLUSION: Wheat-grain moxibustion could significantly improve thyroid function and depression in rats with hypothyroidism. Its mechanism may be related to up-regulating the protein and mRNA expression of MR and GR in the hippocampus, and then affecting the expression of serum cortisol and 5-HT.

[Academic origin of round magnetic needle and standardization operation].

The origin and development of round magnetic needle was explored, and the structure of round magnetic needle was introduced in detail, including the handle, the body and the tip of the needle. The clinical opera tion of round magnetic needle were standardized from the aspects of the methods of holding needle, manipulation skill, tapping position, strength of manipulation, application scope and matters needing attention, which laid foundation for the popularization and application of round magnetic needle.

[Efficacy and safety evaluation of fire needling for rats with rheumatoid arthritis].

OBJECTIVE: To explore the therapeutic effect and safety of fire needling on rat with rheumatoid arthritis. METHODS: Forty female Wistar rats were randomly divided into a normal group, a model group, a methotrexate (MTX) group and a fire needling group, 10 rats in each group. The 0.1 mL of normal saline was subcutaneously injected in rat's right back paw in the normal group, while the other groups received subcutaneous injection of 0.1 mL of complete Freund's adjuvant to establish the model. The pricking method was applied in the fire needling group at "Jiaji" (EX-B 2), "Zusanli" (ST 36) and Ashi points with a depth of 5 mm, once every three days, and totally eight times were required. In the MTX group, with intragastric administration of 2.0 mg/kg per rat, the treatment was given once every seven days, and totally four times were required. The weight, swelling rate of foot, joint pain score and polyarthritis index of rats in each group as well as the pathological change of liver tissue under light microscope were observed. RESULTS: The weight, swelling rate of foot, joint pain score and polyarthritis index in the model group were significantly higher than those in the normal group (P < 0.05, P < 0.01), after the treatment, the above-mentioned indexes in the two treatment groups were obviouly dereased as componed with those in the model group (all P < 0.01), and there were statistical differences between the fire needling group and MTX group (P < 0.05, P < 0.01). Compared with the normal group, inflammatory cells were appeared with a cluster distribution in the liver cells of rats in the model group. The inflammatory cells were reduced in the MTX group and fire needling group, and liver cells had degenerative edema and cell nucleus were largen and deformed in the MTX group. CONCLUSION: The fire needling has significant efficacy for rats with adjuvant arthritis (AA) without any damage to the liver, which coud have a better control of disease progression of rheumatoid arthritis .

[Effect of fire-needle intervention on serum IL-1 and TNF-alpha levels of rheumatoid arthritis rats].

OBJECTIVE: To observe the effect of fire-needle intervention on serum IL-1 and TNF-alpha contents in rheumatoid arthritis (RA) rats so as to study its underlying mechanism in improving RA. METHODS: A total of 40 Wistar rats were randomized into normal control, model, methotrexate (MTX, medication) group and fire-needle groups, with 10 cases in each group. RA model was established by injecting Freund's complete adjuvant (0.1 mL) into the rats' right hind paw. Fire-needle was applied to "Jiaji" (EX-B 2),"Zusanli" (ST 36) and "Ashi point" (the most severe site of the swelling joint), once every three days and 8 times altogether. The rat's right hindpaw swelling volume was detected by volumetric method, and serum IL-1 and TNF-alpha contents were determined by double antibody sandwich ELISA. The pathological changes of synovium tissue of the right knee-joint were observed by optical microscope. RESULTS: In comparison with the control group, the swollen paw volumes at time-points of 12 d, 19 d, 26 d and 33 d after modeling and serum IL-1 and TNF-alpha contents were increased significantly (P < 0.01). Compared to the model group, the swollen paw volumes on day 33 in both medication and fire-needle groups were decreased considerably (P < 0.01), while serum IL-1 and TNF-alpha contents in the medication and fire-needle groups were reduced significantly (P < 0.05, P < 0.01). No significant differences were found between the medication and fire-needle groups in the swollen paw volumes on day 12 (P > 0.05). The pathological states (synovial cells' arrangement, inflammatory infiltration, synovial membrane thickening, etc.) of synovium tissue of the right knee-joint in both medication and fire-needle groups were relatively lighter than those of the model group. CONCLUSION: Fire-needle therapy can down-regulate serum IL-1 and TNF-alpha contents in rheumatoid arthritis rats, and reduce the degree of arthroncus of the affected joint, which may contribute to its effect in improving synovial lesions.

[Therapy of hook needle and manipulations].

With the unique structure and manipulation techniques of blood-letting and cutting, hook needle serves as a role in dredging meridians, removing blood stasis, purging heat for resuscitation and relaxing synechial tissues. There are three needling techniques of hook needle: swift pricking, bleeding and pricking. Six manipulations are commonly used as lifting-thrusting, plucking, pulling, pushing-scraping, vibration and massage. The mechanism and manipulations of hook needling are summarized systematically in the present paper. In this way, its manipulations, indications and contraindications are expounded in detail.

[The history and development of new nine-needle].

This paper introduces the old nine-needle including its name, shape, function, manipulation and origination, as well as systematically introduces the new nine-needle as a new product which has been developed under the guidance of modern science and technology based on the old nine-needle since 1949. As a new tool, the new nine-needle provide much more choice for acupuncture treatment and obtained satisfied clinical effect. During the development of research and clinical practice on the new nine-needle, its theory has been consummated successively. Combining various kinds of needles for treatment is not only paying attention to the specificity of one kind of needles, but also emphasizing the whole function of all kinds of needles, which makes the new nine-needle formed the characteristic and distinct theory of its own.

[Overview of relation between retaining time of acupuncture and effect for apoplexy].

The characteristics and advantages of acupuncture therapy for apoplexy have been widely approved in China. However, the factors affecting the curative effect are various. More and more researchers think highly of the acupuncture needle-retaining duration. The present article makes a discussion on the significance of needle-retaining duration for improving stroke patients' symptoms and signs; and reviews the recent progresses, the extant problems and the academic significance about the studies of the abovementioned topic by combining the authors' previous work. Generally, a systemic study on this topic is rather deficient up to now, and is definitely necessary for further improving clinical therapeutic effect in the light of the requirements of evidence-based medicine.

[Standardized manipulations of sharp hooked needling for treating migraine and investigation on its mechanisms].

A comparison of recognition to migraine and treatment methods for it was made between traditional Chinese medicine and western medicine in this paper. Emphasis was made on standardized manipulation processes and techniques of sharp hooked needling for treating the disease, based on both the organic conception of the human body, the theory held by TCM, and on the principle of treatment according to pathogenesis obtained through differentiation of symptoms and signs. The necessary preparation before operations, methods for its application and announcements in the standardized manipulations of sharp hooked needling were all explained in detail. Based on its needling, pricking, dissecting and relaxing effects, the mechanisms of sharp hooked needling in treating the disease were investigated. It is concluded that sharp hooked needling is very effective in treating migraine.

[Selection of basic acupoints for composing "gastric-disorder-formula" for electroacupuncture prevention of acute gastric mucosal lesion in rats].

OBJECTIVE: To select the best combination of the commonly-used acupoints [Zhongwan (CV 12), Neiguan (PC 6), and Zusanli (ST 36)] for treatment of gastric disorders. METHODS: Forty-eight Wistar rats were randomly divided into 8 groups: model, ST36, CV12, PC6, ST36 + PC6, ST36 + CV12, CV12 + PC6 and ST36 + CV12 + PC6 by factorial design method. Acute gastric mucosal lesion model was established by intragastric perfusion of anhydrous alcohol. EA(1- 3 mA, 30 Hz) was applied to the above-mentioned acupoints for 20 min respectively. And after EA, gastric mucosal lesion index (LI), pathological lesion integral score (PLIS) were calculated and mucosal ultramicrostructural changes observed by optical microscope and transmission electron microscope respectively. RESULTS: Compared with model group, the gastric mucosal LI and PLIS of ST36, CV12, PC6, ST36 + CV12, ST36 + PC6, CV12 + PC6, ST36 + CV12 + PC6 groups all decreased significantly (P < 0.05). Comparison among 7 EA groups showed that both LI and PLIS of ST36 + CV12 + PC6 group were significantly lower than those of ST36, V12, PC6, ST36 + CV12, ST36 + PC6 and CV12 + PC6 groups (P < 0.05). No significant differences were found between every two groups of ST36, CV12, PC6, ST36+ CV12, ST36 + PC6 and CV12 + PC6 groups (P > 0.05) in both LI and PLIS. Ultrastructural results showed that the injury of gastric mucosal parietal cells and delomorphous cells including mitochondrial swelling, crista arrange disorder and breaking, enlargement of the rough endoplasmic reticulum, ribosome exfoliation, etc. in model group was serious, but lightened significantly in EA groups particularly in ST36 + CV12 + PC6 group. CONCLUSION: ST36, PC6 and CV12 combination can effectively reduce anhydrous alcohol-induced gastric mucosal lesion in rats, and may be used as a basic formula

[Effects of electroacupuncture at points "Weibingfang" on EGF and EGFR mRNA expressions in rats with lesion of gastric mucosa].

OBJECTIVE: To probe the mechanism of electroacupuncture at points "Weibingfang" in treatment of acute gastric mucosal lesion. METHODS: Forty Wistar rats of sanitary degree were randomly divided into 4 groups, normal group, model group, sham-model group and Weibingfang group, 10 rats in each group. The acute gastric mucosal lesion model was made by intragastric perfusion of anhydrous alcohol. The Weibingfang group were treated by electroacupuncture at "Neiguan" (PC 6), "Zhongwan" (CV 12) and "Zusanli" (ST 36) with sparse-dense wave, frequency of 10-30 Hz, current intensity of 2 mA, for 20 min. One hour after the treatment, the blood from the abdominal aorta and the gastric mucosa tissue were taken, and serum epidermal growth factor (EGF) level and epidermic growth factor receptor mRNA (EGFR mRNA) expression in the gastric mucosa were detected with enzyme linked immunosorbent assay (ELISA) and reverse transcriptase-polymerase chain reaction (RT-PCR) respectively. RESULTS: There were significant differences in blood EGF level (41.62 +/- 12.58) ng/L and EGFR mRNA expression (0.78 +/- 0.03) in the model group were significantly different from [(60.37 +/- 12.01) ng/L and 0.55 +/- 0.04] in the normal group and [(61.21 +/- 13.46) ng/L and 0.53 +/- 0.05] in the sham-model group (P < 0.05); after electroacupuncture, blood EGF level (70.59 +/- 10.14) ng/L increased and the EGFR mRNA expression (1.18 +/- 0.02) in the gastric mucosa was up-regulated with a significant differences as compared with those in the model group (P < 0.05). CONCLUSION: Point "Weibingfang" can promote proliferation, differentiation and migration of the gastric mucosal epidermic cells to repair the gastric mucosal lesion.

[Survey on the relationship between pathological reactions of acupoints and Zangfu organs/meridians].

In this article, the authors make an analysis on the regularity of correlation between pathological reactions of acupoints and Zangfu organs/meridians, current clinical application and mechanism research, and summarize the relative specificity of the correlation between abnormal reactions of acupoints and states of Zangfu organs/meridians. In addition, the authors also point out some shortcomings in current researches on the above-mentioned aspects, for examples, only observation on the phenomenon without definite conclusion on the regularity of acupoint reactions, being restricted in the objective specificity, etc.. They stress that we should sum up a series of regularities for clinical application (early diagnosis), and conduct profound research on its underlying mechanism in the coming days.

[Effect of electroacupuncture on the contents of excitatory amino acids in cerebral tissue at different time courses in rats with cerebral ischemia and reperfusion injury].

OBJECTIVE: To observe changes of contents of excitatory amino acids in focal cerebral ischemic tissue after electroacupuncture (EA) treatment at different time courses in cerebral ischemia and reperfusion (CI/R) rats so as to provide experimental evidence for selecting the best duration of EA stimulation in the treatment of apoplexy. METHODS: Forty SD rats were randomized into control, model, EA-10 min, EA-30 min and EA-60 min groups. CI/R model was established by middle cerebral artery occlusion (MCAO) and reperfusion. EA (2 Hz, 3.5 mA and continuous waves) was applied to "Baihui" (GV 20) and "Shuigou" (GV 26) for 10 min, 30 min and 60 min respectively. Cerebral glutamate (Glu) and aspartic acid (Asp) concentrations were detected by using an amino acid automatic analyzer after homogenate and centrifugalization of the right brain tissue. RESULTS: Compared with control group, Asp and Glu contents of model group and EA-10 min group increased significantly (P < 0.01); while in comparison with model group, Asp level in EA-60 min group, and Glu level in EA-10 min, EA-30 min and EA-60 min groups decreased considerably (P < 0.05). No significant differences were found between control and EA-30 min groups, and between control and EA-60 min groups in Asp and Glu levels (P > 0.05). CONCLUSION: EA of GV20 and GV26 for 60 min can markedly reduce Glu and Asp levels in the topical cerebral infarct area in CI/R rats, which may contribute to its effect in relieving cerebral ischemic injury. Suitable duration of EA stimulation is definitely necessary for inducing a better therapeutic effect.

[Survey of studies on mechanisms of acupuncture and moxibustion in decreasing blood pressure].

Mechanisms of acupuncture and moxibustion for treatment of hypertension have been studied extensively from poly-aspects, poly-levels and poly-links. The present paper reviews the studies on nervous regulation, humoral regulation and regulation of peripheral vessel resistance and other pathways and their relationships in recent ten years, and indicates further perfecting these studies from the following aspects: studies on the mechanism of section and combination of blood pressure-decreasing points, relationship of time-effect and dose-effect of acupuncture and moxibustion, and studies on mechanisms of functions of nerve-endocrine-immune system.

[Influence of moxibustion on JAK-STAT signal transduction pathways of synovial cells in rheumatoid arthritis rabbits].

OBJECTIVE: To observe the effect of moxibustion on Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway of synovial cells in rheumatoid arthritis (RA) rabbits for revealing the underlying mechanism of moxibustion in the treatment of RA. METHODS: A total of 30 rabbits (Japanese Big-ear White species) were randomized (stratified random) into control, model and moxibustion groups with 10 cases in each. RA model was established by injection of Freund's Complete Adjuvant (FCA, 0.5 mL/kg) into the animal's bilateral joint cavities. Moxibustion was applied to bilateral "Shenshu" (BL 23), 5 cones every time, once daily (except Sundays), 3 weeks altogether. The synovial tissue of joint was sampled for analyzing the expression of signal molecules associated with JAK-STAT pathway with gene chip and bioinformation analytical techniques. RESULTS: Compared with normal control group, the perimeters of both knee joints in model group increased significantly from the 3rd day to 21st day after injection of FCA (P < 0.01), while compared with model group, those in moxibustion group decreased considerably from the 6th day on (P < 0.05, 0.01). In comparison with control group, JAK-STAT pathway-associated genes with up-regulated expression in model group were C/EBP beta, CBP, CRP, GATA3, IFNAR1, IFNAR2, IFNGR2, IL-10Rb, INDO, SH2B, STAT3, STAT6, JAK3 and GP130, and those with down-regulated expression were A2M, MIG and IL-2Rr, suggesting an abnormal activation of JAK-STAT pathway; while in comparison with model group, the related gene up-regulated in moxibustion group in the expression was IL22R and those down-regulated were Cyclin D1, C/EBP beta, CRP, GATA3, IFNAR2, INDO, JAK2, JAK3, V-JUN, STAT3, STAT5, SH2B and OSM, showing that moxibustion had an apparent inhibitory effect on AR-induced abnormal activation of some genes as C/EBP beta, GATA3, IFNAR2, INDO, etc. CONCLUSION: Moxibustion can resist inflammation and eliminate swelling in RA rabbits, which may be closely with its effect in inhibiting abnormal activation of JAK-STAT pathway in synovial cells.