Electroacupuncture Treatment Improves Learning-Memory Ability and Brain Glucose Metabolism in a Mouse Model of Alzheimer’s Disease: Using Morris Water Maze and Micro-PET

Abstract

Introduction. Alzheimer's disease (AD) causes progressive hippocampus dysfunctions leading to the impairment of learning and memory ability and low level of uptake rate of glucose in hippocampus. What is more, there is no effective treatment for AD. In this study, we evaluated the beneficial and protective effects of electroacupuncture in senescence-accelerated mouse prone 8 (SAMP8). Method. In the electroacupuncture paradigm, electroacupuncture treatment was performed once a day for 15 days on 7.5-month-old SAMP8 male mice. In the normal control paradigm and AD control group, 7.5-month-old SAMR1 male mice and SAMP8 male mice were grabbed and bandaged while electroacupuncture group therapy, in order to ensure the same treatment conditions, once a day, 15 days. Results. From the Morris water maze (MWM) test, we found that the treatment of electroacupuncture can improve the spatial learning and memory ability of SAMP8 mouse, and from the micro-PET test, we proved that after the electroacupuncture treatment the level of uptake rate of glucose in hippocampus was higher than normal control group. Conclusion. These results suggest that the treatment of electroacupuncture may provide a viable treatment option for AD.

Full text

Research Article
Electroacupuncture Treatment Improves Learning-Memory
Ability and Brain Glucose Metabolism in a Mouse Model of
Alzheimer’s Disease: Using Morris Water Maze and Micro-PET
Jing Jiang,1Kai Gao,2Yuan Zhou,1Anping Xu,1Suhua Shi,1Gang Liu,3and Zhigang Li1
1Beijing University of Traditional Chinese Medicine, Beijing 100029, China
2Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences & Comparative Medical Center,
Peking Union Medical College, Beijing 100021, China
3Community Health Service Center of Dongcheng District, Beijing 100010, China
Correspondence should be addressed to Zhigang Li; lizhigang@.com
Received  December ; Revised  January ; Accepted  January 
Academic Editor: Mani Vasudevan
Copyright ©  Jing Jiang et al. is is an open access article distributed under the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Introduction. Alzheimer’s disease (AD) causes progressive hippocampus dysfunctions leading to the impairment of learning and
memory ability and low level of uptake rate of glucose in hippocampus. What is more, there is no eective treatment for AD. In this
study, we evaluated the benecial and protective eects of electroacupuncture in senescence-accelerated mouse prone  (SAMP).
Method. In the electroacupuncture paradigm, electroacupuncture treatment was performed once a day for  days on .-month-old
SAMP male mice. In the normal control paradigm and AD control group, .-month-old SAMR male mice and SAMP male
mice were grabbed and bandaged while electroacupuncture group therapy, in order to ensure the same treatment conditions, once a
day,  days. Results. From the Morris water maze (MWM) test, we found that the treatment of electroacupuncture can improve the
spatial learning and memory ability of SAMP mouse, and from the micro-PET test, we proved that aer the electroacupuncture
treatment the level of uptake rate of glucose in hippocampus was higher than normal control group. Conclusion. ese results
suggest that the treatment of electroacupuncture may provide a viable treatment option for AD.
1. Introduction
Alzheimer’s disease (AD) is a progressive neurodegenerative
disease, which is the most widespread cause of dementia
and its incidence will continue to increase rapidly as the
population ages []. It is characterized by the progressive
decline of memory and cognitive function and changes in
behavior and personality []. Despite the fact that extensive
research is focused on AD, there is no eective treatment for
this disease []. erefore, the therapeutic of AD is urgent to
be proposed.
Electroacupuncture (EA) treatment is a type of needling
therapy from Journal of Acupuncture and Moxibustion,num-
ber ,, which combines needling with electric stimula-
tion, connecting needles of the point group concerned (
points make up a group) with pulse current from the electric
stimulator []. ere is a dual-directional pulse current
(intermittent oscillatory current), sin wave, square wave,
and so forth, with characteristics such as continuous wave,
sparse-dense wave, intermittent wave, undulate wave, and
saw tooth wave. e frequency most commonly used is –
 times/sec. over  times/sec. which is used less. What
is more, the proper intensity of the stimulation is marked by
a muscular twitch around the acupoint and a comfortable
sensation []. EA treatment has advantages like many kinds
of oscillation waves, wide range of frequency, and stable
function [].
So far, EA treatment has yielded neuroprotective function
in animal models of depression, spinal cord injury [,],
cerebral ischemia-reperfusion injury [–], stork [,,,
], and many kinds of pain [–]. Clinically, EA treatment
has been shown to have ecacy in curing many kinds of
neurological disease, such as depression [,], spinal cord
injury [–], cerebral ischemia-reperfusion injury [],
stork [–], and many kinds of pain [–]. us, some
of researcher proposed that since the EA treatment could
Hindawi Publishing Corporation
Evidence-Based Complementary and Alternative Medicine
Volume 2015, Article ID 142129, 7 pages
http://dx.doi.org/10.1155/2015/142129

 Evidence-Based Complementary and Alternative Medicine
protect the central nervous system it may be used as an
alternative treatment for AD []. However, more research is
needed to prove this conclusion.
e aim of this study was to assess the ecacy of EA
treatment in curing AD. We utilized the mouse model of
AD, senescence-accelerated mouse prone  (SAMP), which
develops the learning and memory impairment and the mood
disorder. Here, we present the ethology and in vivo imaging
evidence that EA treatment over a period of half a month
improves the learning and memory ability and brain glucose
metabolism in AD, specically in the hippocampus of the
SAMP mouse. is nding extends our previous EA work in
the models of the central nervous injury to demonstrate that
EA treatment is also eective in protecting the brain against
chronicinsultsduetoAD-relateddisease.
2. Method and Materials
2.1. Animals. Senescence-accelerated mouse prone 
(SAMP) and the cognate normal senescence-accelerated
mouse-R (SAMR) breeding pairs were kindly provided
by Professor Takeda at Kyoto University, Japan []. e
animals were housed in a barrier facility of the Experimental
Animal Centre of First Teaching Hospital of Beijing University
of Traditional Chinese Medicine and under live conditions
of controlled temperature ( ±∘C), a  h/ h dark/light
cycle, and sterile drinking water and standard pellet diet
ad libitum. All experiments were performed according to
the National Institute of Health Guide for the Care and Use
of Laboratory Animals (NIH publications number -).
irty .-month-old SAMP male mice were divided into
two groups (𝑛=10per group): SAMR normal control (Rc)
group, SAMP Alzheimer’s disease control (ADc) group, and
SAMP electroacupuncture (EA) group.
2.2. Acupuncture Manipulation. In the EA group, elec-
troacupuncture treatment was performed once a day for 
days (no treatment on the eight day). e prescription of
acupuncture points included DU Baihui,DUShuigou,
and EX-HN Yintang (the signicant extra point). e
locations of these points were according to the National
Acupuncture Society for Experimental Research developed
the “laboratory animal acupuncture atlas”. Huatuo card ,
. inch needle was used for treatment. Pricking method was
usedforDUShuigou;atthornmethodwasusedfor
DU Baihui and EX-HN Yintang. Needle depth was . cm
and taped. e needle handle was connected with HANS-
LH electroacupuncture device (Peking University Institute
of Science Nerve and Beijing Hua Wei Industrial Development
Company), sparse wave,  Hz of the frequency,  V of the
voltage, and . mA of the current intensity.
In the Rc group and ADc group, do not do any treatment
under the same rearing conditions, while grabbing and
bondage the mice in order to ensure the same treatment
conditions, once a day,  days.
2.3. Morris Water Maze Behavioral Test. e Morris water
maze consisted of a circular tank ( cm in diameter,  cm in
height) lled with water to a depth of  cm maintained at 
±∘C and rendered opaque with blue-black ink. A removable
circular platform (. cm diameter,  cm height) with its top
surface  cm below the water was located inside the pool. e
area of the pool was conceptually divided into four quadrants
(NE, NW, SW, and SE) of equal size. Data were collected by
a video camera (TOTA-d,Japan)whichwasxedtothe
ceiling of the room and connected to a video recorder and an
automated tracking system (China Daheng Group, Beijing,
China).
In this behavioral test, mice are placed in the pool of water
containing a platform just below the surface of the water. ey
escape from the maze when they nd the platform. Distal
visual cues are arrayed around the room, and in general, mice
are able to learn the location of the hidden platform based on
these cues.
2.4. Hidden Platform (Place) Testing. is portion of the test
assesses the ability of the mice to nd the platform under
conditions where they cannot directly see it but must either
remember it is relative to external cues or perform a search
for it. e platform was placed cm under the surface of the
water, and the water was opaque by a suspension of dark
blue, nontoxic tempera paint. e platform was placed in
a dierent location from that used in the visible platform
testing. Each mouse was released from one of  locations and
had  s to search for the hidden platform. At the end of each
trail,themousewasplacedontheplatformorallowedtostay
on the platform for  s. Prominent spatial cues were arrayed
around the room. e investigator is also a powerful spatial
cueandalwayssatinthesamelocationduringeachtrailaer
releasing the mouse. Eight trails per day for  consecutive
days were performed with the location of the platform kept
constant. We recorded the time that the mouse found the
platform needed, and we call it escape latency.
2.5. Probe Trail. e day aer the completion of hidden
platformtesting,theplatformwasremoved,andeachmouse
was placed in the pool once for  s, starting from the same
starting location as was used rst in hidden platform testing.
e time spent swimming in the quadrant where the platform
had been was recorded. is is considered to be the most
specic test for spatial memory. We recorded the time that
themousespentintheplatformquadrantandcalculatedthe
percentage of total time spent in swimming to the platform
quadrant.
2.6. Micropositron Emission Tomography. Before experi-
ments, each mouse (. months, ∼ g) for blood glucose
monitoring, the results showed the normal range (.∼
. mmol/L) could be used for micro-PET detective (18F-
FDG PET tracer was provided by the Chinese Medicine
Research Institute PET Room; PET imaging system using
Siemens INVEON PET/CT imaging system). Six hours of
water deprivation before the experiment. e mice were
placed in the suction chamber, inhaling the oxygen mixed
with.%isouranetobeanesthetized.Aercompleteanes-
thesia, approximately .∼. MBq 18F-FDG PET tracers

Evidence-Based Complementary and Alternative Medicine 
were injected via vena of tail. Aer the 18F-FDG PET tracer
uptake for  min, the mice were placed on the scan bed in
prone position, the mice and scanner long axis were parallel,
andtheheadofmousewaslocatedwithinthescanner
eld of view. en the micropositron emission tomography
began to collect the image. During this progress, the mice
were anesthetized by the oxygen mixed with .% isourane
( L/min).
2.7. Micropositron Emission Tomography Image Reconstruc-
tion. Filtered back projection (FBP) and CT photon attenua-
tion correction were used for image reconstruction. Dynamic
micro-PET image frames are taken  s/frames.
2.8. Region of Interest Selection. e three-dimensional
region of interest te chnology was applied for manual selection
of the hippocampus three-dimensional region of interest in
transverse, coronal, and sagittal planes. en calculate the
uptake rate of per gram with the region of interest.
2.9. Statistical Method. AlldatawereanalyzedbySPSS(ver-
sion .; SPSS, Inc., Chicago, IL, USA). All measurements
were performed by an independent investigator blinded to
the experimental conditions. e results in the gures are
expressed as the mean ±standard deviation. Dierences
within or between normal distributed data were analyzed by
analysis of variance (ANOVA) followed by Huynh-Feldt test
(for Morris water maze test). Statistical signicance was set at
𝑃 < 0.05.
3. Results
3.1. Eect of Electroacupuncture in Spatial Learning Ability
of SAMP8 Mouse in the Morris Water Maze Test. e eect
of electroacupuncture in spatial location ability of SAMP
mouseintheWMWtestiselucidatedinFigure (a).Wecan
see that with the training time extension, the escape latency
of all groups had shown a downward trend (Figure (b)). e
AD control group showed marked retardation in the escape
latency, probably due to the memory decits resulting from
the rapid aging process impairment of learning and memory.
e analysis of the escape latency revealed that the mouse
in EA group had signicantly reduced the escape latency
compared with the AD control group (𝑃 < 0.05,Figure (c)).
3.2. Eect of Electroacupuncture in Spatial Memory Ability of
SAMP8 Mouse in the Morris Water Maze Test. To investigate
the eect on spatial memory ability, the performance in
theprobetrialondaywasexaminedbyanalyzingthe
percentages of time spent swimming to the expected position
of the platform. A higher percentage of time spent in the
platform quadrant is interpreted as a higher level of memory
retention []. In this probe trial, we found that compared
with AD control group, EA group spent higher time in the
platform quadrant (𝑃 < 0.01). What is more, in percentage of
time spent in the platform quadrant, EA group and normal
group had no signicant dierence in statistics (𝑃 = 0.223,
Figure ).
3.3. PET Imaging of Mice Hippocampus. Because of the eects
of the tail vein injection, condition of anesthesia, and the
metabolism of the 18F-FDG, four animals of each group could
successfully guarantee the completion of micro-PET test.
Use the same color standard and color code from top
high to the bottom low to display the metabolic rate of the
glucose. e le of the observer is the right of the animal.
Fromtheimage,aerthetreatmentofelectroacupuncturethe
18F-FDG of hippocampus is higher than Alzheimer’s disease
group (Figure ).
3.4. 18F-FDGUptakeRateofperGraminHippocampusTissue.
To study that the treatment of electroacupuncture corre-
sponds to enhancing the glucose metabolic activity in hip-
pocampus, 18F-FDGPETscanwasperformedonthemice.
e result showed that aer treatment of electroacupuncture
theuptakerateof18F-FDG in hippocampus was higher
than Alzheimer’s disease group and normal control group
(Figure ).
4. Discussion
In the current research, we studied the eect of elec-
troacupuncture on animal model of Alzheimer’s disease using
Morriswatermazeandmicro-PETandaimedtondthat
whether the treatment of electroacupuncture can improve
the condition of Alzheimer’s disease. Using the Morris water
maze,wefoundaerthetreatmentofelectroacupuncturethat
the spatial learning and memory ability of the SAMP mouse
had improved. Further, the result of micro-PET revealed that
treatment of electroacupuncture can increase the uptake rate
ofglucoseinhippocampusofSAMPmouse.esendings
from animal behavior and in vivo imaging lead us to conclude
that the treatment of electroacupuncture may play a curable
role in Alzheimer’s disease, particularly in the learning and
memory ability.
4.1. Alzheimer’s Disease in Traditional Chinese Medicine and
the Acupuncture Prescription. Alzheimer’s disease belonged
to encephalopathy in Chinese medicine. It is caused by de-
ciency of jing and blood, with the aged condition, leading to
serious brain function disorder. It is characterized by forgetful
and personality changes []. In the treating principle of
Chinese medicine, according to the principle “the brain is
the house of mentality,” “the Governor Vessel ... entering
the brain and extending up to the very top of the head,”
based on the close relationship brain-mentality and brain-
Governor Vessel, we proposed “dredging the Governor Vessel
andawakeningmind”methodtotreatAlzheimer’sdisease.
In the selection of acupoints, we chose DU Baihui,DU
 Shuigou, and EX-HN Yintang as the main points. DU
Baihui, a meridian point of the Governor Vessel, and the
meeting point of Governor Vessel, the three Yang Meridians
ofthehandandfoot,fromtheA-BClassicofAcupuncture
and Moxibustion (zhen jiu jia yi jing ), also named Sanyang-
wuhui,Dianshang,Wu hui.Itislocatedonthehead,cun
directly above the midpoint of the anterior hairline. Its
indication is headache, dizziness, palpitation due to fright,

 Evidence-Based Complementary and Alternative Medicine
Group Day 1 Day 2 Day 3 Day 4 Day 5
Normal control group
Alzheimer’s disease
control group
Electroacupuncture
group
41.85 ± 6.32 32.88 ± 8.78 27.98 ± 10.92 24.82 ± 6.64 19.21 ± 12.63
57.21 ± 3.68 57.26 ± 5.09 54.70 ± 7.57 53.16 ± 8.16 53.67 ± 8.28
57.65 ± 4.15 51.25 ± 9.08 49.42 ± 14.01 48.32 ± 10.25 47.10 ± 10.78
(a)
80
60
40
20
0
Escape latency (s)
Day 1Day 2Day 3Day 4Day 5
Days
Normal control group
Alzheimer’s disease group
Electroacupuncture group
(b)
P < 0.05
Normal
control group
Alzheimer’s
disease group
Electroacupuncture
group
Groups
80
60
40
20
0
Escape latency (s)
(c)
F : (a) Comparison of the escape latency of all groups. (b) e trend of the escape latency of all groups. (c) Comparison the mean of
the escape latency of all groups.
30
20
10
0
Normal
control group
Alzheimer’s
disease group
Electroacupuncture
group
Groups
e time spent in
platform quadrant (%)
(a)
P < 0.01
Normal
control group
Alzheimer’s
disease group
Electroacupuncture
group
Groups
30
20
10
0
e time spent in
platform quadrant (%)
(b)
F : (a) e percentage of time spent in platform quadrant of each group. (b) e percentage of time spent in platform quadrant of each
group.
amnesia, corpse-like syncope, aphasia from apoplexy, manic-
depressive psychosis, epilepsy, hysteria, and so forth. DU 
Shuigou is a point on the Governor Vessel, Hand-Yangming,
and Foot-Yangming, also called Renzhong. Its location is on
theface,atthejunctionoftheupper/andmiddle/of
the philtrum. It is used for coma, syncope, manic-depressive
disorder,epilepsy,acuteandchronicinfantileconvulsion,and
so forth. EX-HN Yintang, an extra point, is seen in Bian
Que’s Jade Dragon Classics of Acupuncture and Moxibustion
(Bian Que Shenying Zhenjiu Yulong Jing). It is on the forehead,
at the midpoint between the eyebrows. Headache, vertigo,
insomnia, and puerperal faintness are the indication of EX-
HN.Ingeneral,weusetheabovethreeacupointsasthe
acupuncture prescription.
4.2. Senescence-Accelerated Mouse-P8 (SAMP8) Is an Optional
Animal Model for Alzheimer’s Disease. e senescence-
accelerated mouse (SAMP) is a spontaneous animal model
of Alzheimer’s disease, and it develops early memory distur-
bances and changes in the blood-brain barrier resulting in
decreased eux of amyloid-beta protein from the brain [].
is nontransgenic animal model with great utility can be
better simulated for the memory decits and the low level
uptakerateofglucoseinhippocampus[]. So, in the current
research, this kind of animal model can help us to nd the
eect of electroacupuncture treatment in curing Alzheimer’s
disease.
4.3. Eect of Electroacupuncture in Morris Water Maze. e
Morris water maze (MWM) is one of the most common tasks

Evidence-Based Complementary and Alternative Medicine 
6
0
(a)
6
0
(b)
6
0
(c)
F : Right side hippocampus of the mice in micro-PET scan image. (a) Normal control group; (b) Alzheimer’s disease group; (c)
electroacupuncture treatment group. Color code: min = , max = .
5
4
3
2
1
0
ID/g (%)
Normal
control group
Alzheimer’s
disease group
Electroacupuncture
group
Groups
(a)
6
4
2
0
P < 0.01
Normal
control group
Alzheimer’s
disease group
Electroacupuncture
group
Groups
ID/g (%)
(b)
F:(a)euptakerateof18F-FDG per gram in hippocampus. (b) e uptake rate of 18F-FDG per gram in hippocampus.
used to assess spatial learning and memory ability in rodents
[]. Spatial navigation performance in the hidden goal task
(HGT), a real-space human analogue of the Morris water
maze, can identify mild cognitive impairment (MCI) patient
with memory impairment of the hippocampus type, a known
indicator of incipient Alzheimer’s disease []. In our study,
we found that aer the treatment of electroacupuncture, the
spatial learning and memory ability of the SAMP mouse had
improved compared with the nontreatment SAMP mouse,
which suggested that the electroacupuncture may improve
the cognitive ability of Alzheimer’s disease patients.
4.4. Eect of Electroacupuncture in Micro-PET. Studies sug-
gested that the cognitive impairment of Alzheimer’s disease
to a certain extent results from the low level of uptake rate
of glucose in hippocampus []. So in our current study, we
used the micro-PET to get the in vivo image of the uptake rate
of glucose in hippocampus.
Positron emission tomography is a noninvasive func-
tionalbrainimagingtechniqueatthemolecularlevel,
which makes the use of radioactive marker to analyze the
metabolism condition in the brain, images the distribution of
biologically targeted radiotracer with high sensitivity []. It
can directly reect the activity of neurons, which becomes an
important tool for diagnosing disease and evaluating ecacy
[]. With the growing importance of animal research in
modern molecular biology, the appearance of micropositron
emission tomography (micro-PET) makes the possible of
in vivo molecular imaging. Development of micro-PET
instrumentation for small animal imaging and the availability
of positron-emitting tracers have made this technology acces-
sible for the noninvasive, quantitative, and repetitive imaging
of biological function in living animals. e development
of new probes and positron-imaging based reporter genes
has extended micro-PET applications to investigations of
metabolism, enzyme activity, receptor-ligand interactions,
protein-protein interactions, gene expression, adoptive cell
therapy, and somatic gene therapy [].
In this research, the 18F-FDG uptake condition in the
hippocampus of mice was imaged by micro-PET, which can

 Evidence-Based Complementary and Alternative Medicine
show the metabolism level in the hippocampus of Alzheimer’s
disease mice. Seeing from the images, the 18F-FDG uptake
condition of electroacupuncture treatment group is higher
than Alzheimer’s disease group. With further calculation and
comparison of the 18F-FDG uptake rate of each group, we can
see that the electroacupuncture treatment group is the high-
est, normal control group is in the middle, and Alzheimer’s
diseasegroupisthelowest.eaboveresultsshowed,aerthe
electroacupuncture treatment the glucose metabolism level
in the hippocampus of the Alzheimer’s disease animal model
would be higher. erefore, we could draw the conclusion
that the treatment of electroacupuncture could improve the
level of uptake rate of glucose in hippocampus in Alzheimer’s
disease animal.
5. Conclusion
In this research, using the test of Morris water maze and
the micro-PET in Alzheimer’s disease animal model SAMP
mouse, we found that the treatment of electroacupuncture
can improve the spatial learning and memory ability by
heightening the level of uptake rate of glucose in hippocam-
pus. is is an interesting notion; however, further research
is needed to prove.
Conflict of Interests
e authers declare that there is no conict of interests
regarding the publication of this paper.
Acknowledgments
is research was supported by National Natural Science
Foundation of China (no. ).
References
[] C. Ballard, S. Gauthier, A. Corbett, C. Brayne, D. Aarsland, and
E. Jones, “Alzheimer’s disease,” e Lancet,vol.,no.,pp.
–, .
[]K.Blennow,M.J.deLeon,andH.Zetterberg,“Alzheimer’s
disease,” e Lancet,vol.,no.,pp.–,.
[] X. T. Sun, L. Jin, and P. X. Ling , “Review of drugs for Alzheimer’s
disease,” Drug Discoveries & erapeutics,vol.,no.,pp.–
, .
[] G.A.Ulett,S.Han,andJ.-S.Han,“Electroacupuncture:mech-
anisms and clinical application,” Biological Psychiatry,vol.,
no. , pp. –, .
[] Y.Fukazawa,T.Maeda,andS.Kishioka,“epharmacological
mechanisms of electroacupuncture,” Current Opinion in Investi-
gationalDrugs(London,England:2000),vol.,no.,pp.–,
.
[]G.A.Ulett,“Conditionedhealingwithelectroacupuncture,”
Alternative erapies in Health and Medicine,vol.,no.,pp.
–, .
[] J.-W. Yang, S.-M. Jeong, K.-M. Seo, and T.-C. Nam, “Eects of
corticosteroid and electroacupuncture on experimental spinal
cord injury in dogs,” Journal of Veterinary Science (Suwon-si,
Korea),vol.,no.,pp.–,.
[] H. Sumano, E. Bermudez, and K. Obregon, “Treatment of
wobbler syndrome in dogs with electroacupuncture,” Deutsche
Tierarztliche Wochenschri,vol.,no.,pp.–,.
[]Y.Zhang,H.Xu,H.Sun,S.Chen,andF.Wang,“Elec-
troacupuncture treatment improves neurological function asso-
ciated with regulation of tight junction proteins in rats with
cerebral ischemia reperfusion injury,” Evidence-Based Comple-
mentary and Alternative Medicine,vol.,ArticleID,
 pages, .
[] C. Wang, F. Yang, X. Liu, M. Liu, Y. Zheng, and J. Guo, “Neu-
rotrophic signaling factors in brain ischemia/reperfusion rats:
dierential modulation pattern between single-time and mul-
tiple electroacupuncture stimulation,” Evidence-Based Comple-
mentary and Alternative Medicine, vol. , Article ID ,
 pages, .
[] F. Guo, W. Song, T. Jiang et al., “Electroacupuncture pretreat-
ment inhibits NADPH oxidase-mediated oxidative stress in
diabetic mice with cerebral ischemia,” Brain Research, vol. ,
pp.–,.
[] F. Guo, T. Jiang, W. Song et al., “Electroacupuncture attenuates
cerebral ischemia-reperfusion injury in diabetic mice through
adiponectin receptor -mediated ohosphorylation of GSK-𝛽,”
Molecular Neurobiology,.
[] F.Tan,J.Chen,Y.Liangetal.,“Electroacupunctureattenuates
cervical spinal cord injury following cerebral ischemia/reperfu-
sion in stroke-prone renovascular hypertensive rats,” Experi-
mental and erapeutic Medicine,vol.,no.,pp.–,
.
[] W. S. Wang, W. Z. Tu, R. D. Cheng et al., “Electroacupuncture
andA-depressthetransmissionofpainonprimary
aerent mediated by the PX3receptor in rats with chronic
neuropathic pain states,” Journal of Neuroscience Research,vol.
, no. , pp. –, .
[] K. Liu, X.-Y. Gao, L. Li et al., “Neurons in the nucleus tractus
solitarius mediate the acupuncture analgesia in visceral pain
rats,” Autonomic Neuroscience,vol.,pp.–,.
[] H.Li,S.Hu,J.Zhangetal.,“Eectsandmechanismsofauricular
electroacupuncture on visceral pain induced by colorectal
distension in conscious rats,” Acupuncture in Medicine,vol.,
no. , pp. –, .
[] Y.-H. Gao, J.-Y. Wang, L.-N. Qiao et al., “NK cells mediate the
cumulative analgesic eect of electroacupuncture in a rat model
of neuropathic pain,” BMC Complementary and Alternative
Medicine,vol.,no.,p.,.
[] X.- M. Chen, J. Xu, J.-G. Song, B.-J. Zheng, and X.-
R. Wang, “Electroacupuncture inhibits excessive interferon-
gamma evoked up-regulation of PX receptor in spinal
microglia in a CCI rat model for neuropathic pain,” British
Journal of Anaesthesia,vol.,no.,pp.–,.
[] W.-T. Chen, F.-C. Chang, Y.-H. Chen, and J.-G. Lin, “An
evaluation of electroacupuncture at the Weizhong acupoint
(BL-) as a means of relieving pain induced by extracorporeal
shock wave lithotripsy,” Evidence-Based Complementary and
Alternative Medicine,vol.,ArticleID,pages,.
[] Z.Y.Ju,H.S.Cui,X.H.Guo,H.Y.Yang,J.S.He,andK.Wang,
“Molecular mechanisms underlying the eects of acupuncture
on neuropathic pain,” Neural Regeneration Research,vol.,no.
, pp. –, .
[] Z. J. Zhang, R. Ng, S. C. Man et al., “Use of electroacupuncture
to accelerate the antidepressant action of selective serotonin
reuptake inhibitors: a single-blind, randomised, controlled

Evidence-Based Complementary and Alternative Medicine 
study,” Hong Kong Medical Journal, vol. , supplement , pp.
–, .
[]D.K.Weiner,C.G.Moore,N.E.Morone,E.S.Lee,andC.
Kent Kwoh, “Ecacy of periosteal stimulation for chronic pain
associated with advanced knee osteoarthritis: a randomized,
controlled clinical trial,” Clinical erapeutics,vol.,no.,pp.
.e–.e, .
[] M.L.Yeh,Y.C.Chung,K.M.Chen,M.Y.Tsou,andH.H.Chen,
“Acupoint electrical stimulation reduces acute postoperative
pain in surgical patients with patient-controlled analgesia: a
randomized controlled study,” Alternative erapies in Health
and Medicine,vol.,no.,pp.–,.
[] Y.X.Chen,K.M.Kong,W.D.Wang,C.H.Xie,andR.H.
Wu, “Functional MR imaging of the spinal cord in cervical
spinal cord injury patients by acupuncture at LI  (Hegu) and
LI (Quchi),” in Proceedings of the 29th Annual International
Conference of IEEE-EMBS, Engineering in Medicine and Biology
Society (EMBC ’07), pp. –, August .
[] A. M. K. Wong, C.-P. Leong, T.-Y. Su, S.-W. Yu, W.-C. Tsai, and
C. P. C. Chen, “Clinical trial of acupuncture for patients with
spinal cord injuries,” eAmericanJournalofPhysicalMedicine
& Rehabilitation,vol.,no.,pp.–,.
[]H.Zhang,T.Kang,L.Li,andJ.Zhang,“Electroacupuncture
reduces hemiplegia following acute middle cerebral artery
infarction with alteration of serum NSE, S-B and endothe-
lin,” Current Neurovascular Research,vol.,no.,pp.–,
.
[] J. Sun, H. Sang, and C. Yang, “Electroacupuncture improves
orthostatic tolerance in healthy individuals via improving car-
diac function and activating the sympathetic system,” Europace,
vol. , no. , pp. –, .
[] K.-W. Yu, C.-L. Lin, C.-C. Hung et al., “Eects of elec-
troacupuncture on recent stroke inpatients with incomplete
bladder emptying: a preliminary study,” Clinical Interventions
in Aging,vol.,pp.–,.
[] W. T. Hsing, M. Imamura, K. Weaver, F. Fregni, and R. S.
Azevedo Neto, “Clinical eects of scalp electrical acupuncture
in stroke: a sham-controlled randomized clinical trial,” e
Journal of Alternative and Complementary Medicine,vol.,no.
, pp. –, .
[] S. P. Zhang, T. T. Chiu, and S. N. Chiu, “Long-term ecacy
of electroacupuncture for chronic neck pain: a randomised
controlled trial,” Hong Kong Medical Journal,vol.,supplement
, pp. –, .
[] B.-J. Wan, W. Huang, Y.-X. Zhang, and H.-S. Zhang , “Inuence
of electroacupuncture with penetration needling method on
comprehensive pain score in patients with cervical spondylotic
radiculopathy,” Chinese Acupuncture & Moxibustion,vol.,no.
, pp. –, .
[] Z.-R. Sun, J.-H. Yue, and Q.-H. Zhang, “Electroacupuncture at
Jing-jiaji points for neck pain caused by cervical spondylosis: a
study protocol for a randomized controlled pilot trial,” Tri als ,
vol.,no.,article,.
[] S. Mucuk, M. Baser, and T. Ozkan, “Eects of noninvasive elec-
troacupuncture on labor pain, adrenocorticotropic hormone,
and cortisol,” Alternative erapies in Health and Medicine,vol.
,no.,pp.–,.
[] S. Lee, J.-H. Kim, K.-M. Shin et al., “Electroacupuncture to treat
painful diabetic neuropathy: study protocol for a three-armed,
randomized, controlled pilot trial,” Tri als ,vol.,article,
.
[] C. C. L. Xue, R. D. Helme, S. Gibson et al., “Eect of elec-
troacupuncture on opioid consumption in patientsw ithchronic
musculoskeletal pain: protocol of a randomised controlled
trial,” Tr i als ,vol.,article,.
[] K.-F. Chung, W.-F. Yeung, Z.-J. Zhang et al., “Randomized non-
invasive sham-controlled pilot trial of electroacupuncture for
postpartum depression,” Journal of Aective Disorders,vol.,
no. –, pp. –, .
[] Y. O. Cakmak, “A review of the potential eect of elec-
troacupuncture and moxibustion on cell repair and survival: the
role of heat shock proteins,” Acupuncture in Medicine,vol.,no.
, pp. –, .
[] J.E.Morley,H.J.Armbrecht,S.A.Farr,andV.B.Kumar,“e
senescence accelerated mouse (SAMP) as a model for oxida-
tive stress and Alzheimer’s disease,” Biochimica et Biophysica
Acta,vol.,no.,pp.–,.
[] K. Bromley-Brits, Y. Deng, and W. Song, “Morris water maze
test for learning and memory decits in Alzheimer’s disease
model mice,” Journal of Visualized Experiments,no.,article
, .
[] Z. B. Wang Yongyan, Encephalopathy of Chinese Medicine,
People’s Health Publishing House, Beijing, China, .
[] D. S. Woodru-Pak, “Animal models of Alzheimer’s disease:
therapeutic implications,” Journal of Alzheimer’s Disease,vol.,
no. , pp. –, .
[]J.F.Ge,C.C.Qi,J.P.Qiao,C.W.Wang,andJ.N.Zhou,
“Sex dierences in ICR mice in the morris water maze task,”
Physiological Research,vol.,no.,pp.–,.
[] J. Lacz´
o, R. Andel, M. Vyhnalek et al., “From morris water
maze to computer tests in the prediction of Alzheimer’s disease,”
Neurodegenerative Diseases,vol.,no.–,pp.–,.
[] L. Mosconi, R. Mistur, R. Switalski et al., “FDG-PET changes in
brain glucose metabolism from normal cognition to pathologi-
cally veried Alzheimer’s disease,” European Journal of Nuclear
Medicine and Molecular Imaging,vol.,no.,pp.–,
.
[]B.Foster,U.Bagci,A.Mansoor,Z.Xu,andD.J.Mollura,
“A review on segmentation of positron emission tomography
images,” Computers in Biology and Medicine, vol. , pp. –,
.
[] S. E. Schindler, J. McConathy, B. M. Ances, and M. I. Diamond,
“Advances in diagnostic testing for Alzheimer disease,” Missouri
Medicine,vol.,no.,pp.–,.
[] H. R. Herschman, “Micro-PET imaging and small animal
models of disease,” Current Opinion in Immunology,vol.,no.
, pp. –, .