Mediation of the ulnar nerve in the inhibitory effects of HT7 stimulation on cocaine-induced locomotion.

<p>A: The inhibitory effects of HT7 on cocaine locomotion are completely blocked by severing the ulnar nerve (Ulnar+HT7), but not by radial and median nerves (Rad/Med+HT7) (Con, n=12; HT7, n=12; Ulnar+HT7, n=11; Rad/Med+HT7, n=9). Data are analyzed by one way ANOVA with Holm-Sidak post hoc comparison. B: Mechanical stimulation of needle inserted into the tip of the 5^th digit produced a similar effect). Data are presented as mean ± SEM and analyzed by two-way repeated ANOVA with Holm-Sidak post hoc comparison. * indicates significance at p≤ 0.05.</p

The effects of acupuncture at HT7 require large A-fiber activation of ulnar nerve.

<div><p>A: HT7 effects are not blocked by a perineural injection of RTX (resiniferatoxin; a C/Aδ-fiber inhibitor) into ulnar nerve (RTX+HT7). B: Direct injection of capsaicin into HT7 to stimulate C/Aδ-fibers failed to produce HT7 effects (Capsaicin in HT7). *P<0.05 compared to control (cocaine only). n= 6-8 rats for each group of A-B. Data are presented as mean ± SEM and analyzed by one or two-way repeated ANOVA with Holm-Sidak post hoc comparison.</p> <p>C, D: Epifluorescent images showing IB4-labelled neurons in C8 DRG of a rat injected with IB4 into RTX- and vehicle-treated ulnar nerve. An FITC IB4 tracer (FITC-conjugated isolectin B4) was administered into ulnar nerve on the both side of rats, 48 hours after perineural injection of either RTX or vehicle into right and left ulnar nerve, respectively. Numerous IB4-labelled cells of small size are shown in vehicle-treated DRG (C), whereas a few IB4-labelled neurons are found in the RTX-treated DRG (D). It indicates that retrograde labeling of C-fibers by FITC IB4 tracer is blocked by perineural injection of RTX, but not vehicle. Bar=100 μm. </p></div

Mechanical acupuncture by MAI decreases cocaine-induced locomotor activity in a stimulus time-dependent manner.

<p>A: Effect of manual acupuncture at HT7 by 4 different operators (#1 to #4) on cocaine-induced locomotor activity. Acupuncture was performed by 4 operators consisting of 2 beginners (#1, #2) and 2 well-trained operators with experience over 2 years in acupuncture experiments (#3, #4). Variations in the effectiveness of acupuncture on cocaine locomotion are seen among experimenters. B, C: Effect of mechanical stimulation applied to HT7 with MAI on cocaine locomotion. Acupuncture needles were mechanically stimulated at different times (0, 10, 20 or 40 sec) with MAI (1.3 m/sec² in intensity and 85 Hz in frequency). The acupuncture-induced inhibition of cocaine locomotion was stimulus-time dependent, reaching a maximum at 20-sec duration (B). Two different intensities (0.9 or 1.3 m/sec²) were applied for 20 sec to the needles inserted at a depth of 3 mm into HT7. The effect was produced with two different intensities of mechanical stimulation, either 0.7 (low) or 1.3 (high) m/sec² at 20-sec duration, compared to control (cocaine only) (C). D: Effect of mechanical acupuncture at control point LI5 with MAI on cocaine locomotion. No acupuncture effect was produced when a parameter of 20-sec duration and 1.3 m/sec² intensity was applied to LI5 acupoint at 85 Hz. n= 6-8 rats for each group. Data are presented as mean ± SEM and analyzed by two-way repeated ANOVA with Holm-Sidak post hoc comparison. *p<0.05 compared to control (cocaine only).</p

Possible involvement of mechanoreceptors, Pacinian and Meissner corpuscles, in HT7 effects.

<p>A: The HT7 effects are generated by mechanical stimulation applied to HT7 at either high (200 Hz) or low (50 Hz) frequencies. *p<0.05 compared to control (cocaine only). n= 6-8 rats for each group. B: A diagrammatic representation of the proposed hypothesis that the effects of manual acupuncture at HT7 on cocaine-induced locomotion are mediated via A-fiber activation of ulnar nerve originating from superficial and deep tissue. Data are presented as mean ± SEM and analyzed by two-way repeated ANOVA with Holm-Sidak post hoc comparison. </p

Mechanical acupuncture instrument (MAI).

<p><b>A</b>, <b>B</b>: In traditional manual acupuncture, a needle is manually stimulated by manipulating the handle of the needle with an index finger after being inserted into acupoints (A). Manipulating a needle (arrows) elicited abrupt increases of intensity, but with considerable variations in peak values of intensity (B). <b>C</b>, <b>D</b>: With the novel MAI, a cell phone motor is connected to the acuneedle to create a constant vibration of the needle and operated by a control unit for adjustment of intensity (acceleration), frequency and operation time (C). The acceleration parameter of 1.3 m/sec² in intensity and 85 Hz in frequency was routinely used throughout our experiments (D). </p

Response of A- or C-fibers during MAI stimulation at HT7.

<div><p>A-C: Somatic afferent responses of Aβ-fibers (n=14) during MAI stimulation of HT7. A, B: A representative single-unit recording (A) and frequency histogram (B) of response to HT7 MAI stimulation (10 sec) in Aβ-fibers. Frequencies of discharge activity dramatically increased during MAI stimulation of HT7, compared to values before MAI stimulation (C).</p> <p>D-F: Somatic afferent responses of C-fibers (n=10) during MAI stimulation of HT7. A, B: A representative single-unit recording (D) and frequency histogram (E) of response to HT7 MAI stimulation (10 sec) in C-fibers. Frequencies of discharge activity significantly increased during MAI stimulation of HT7, compared to values before MAI stimulation (F).</p> <p>Data are presented as mean ± SEM and analyzed by paired Student’s t-test. *p<0.05 compared to baseline (before HT7-MAI).</p></div

Mediation of peripheral afferents in the inhibitory effects of acupuncture at HT7 on cocaine locomotion.

<p>A: Effect of pretreatment with the local anesthetic bupivacaine on the effects of acupuncture at HT7. Inhibition of cocaine locomotion following mechanical stimulation to HT7 was prevented by infiltration around acupoints with bupivacaine. (*P<0.05 vs. Con group (cocaine only); ^#P<0.05 vs. Saline+HT7 group; Con, n=7; Saline+HT7, n=12; Bupivacaine+HT7, n=12). B: Effect of needling to HT7 at different depths on cocaine locomotion. The HT7 effects are produced by either superficial (1 mm) or deep (3 mm) tissue stimulation (*P<0.05 vs. control; Con, n=11; HT7-1mm, n=12; HT7-3mm, n=12). Data are presented as mean ± SEM and analyzed by two-way repeated ANOVA with Holm-Sidak post hoc comparison.</p

Enhanced c-Fos expression in DiI-labeled DRG neurons following HT7-MAI stimulation.

<p>A-C: Immunofluorescent images of the expression of c-Fos (B) in DiI-labeled DRG neurons (A) from control group (no stimulation at HT7). D-F: The expression of c-Fos (E) in DiI-labeled DRG neurons (D) following MAI stimulation at HT7. The numbers of c-Fos positive cells were markedly increased (E) and double labeled cells of c-Fos/DiI were found in the DRG following HT7-MAI stimulation (F), whereas few c-Fos positive cells were seen in untreated DRGs (B). Bar=100 μm.</p

Spearman's rank correlation maps between the mean SIRs of each layer in cortices.

<p>(A) Correlation map of Aud for NOSTIM (B) Correlation map of Aud for STIM (C) Correlation map of Sens for NOSTIM (D) Correlation map of Sens for STIM (E) Correlation map of Vis for NOSTIM (F) Correlation map of Vis for STIM. The correlation coefficient, ρ, is indicated with color maps ranging from blue to red; blue and red indicates the weakest and strongest correlation, respectively. A strong correlation between two layers indicates that the manganese accumulations in the two are likely to be linearly proportional to each other.</p

Comparison of SIRs between STIM and NOSTIM in the brain regions of auditory and olfactory pathways.

<p>Mann-Whitney <i>U</i>-tests were used to compare the mean SIRs of brain regions. The <i>z</i>-values were calculated from Mann-Whitney's <i>U</i>-values and their standard deviations. SIR is the normalized signal intensity of each ROI to its adjacent Temporalis muscles.</p><p>*<i>P</i><0.05.</p