• Journal of Acupuncture Research
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• v.25 no.3
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• pp.179-187
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• 2008

Objectives & Methods : This study was to investigate effect of low frequency electroacupuncture on NADPH-d positive neurons in the brain cortex of rat with adjuvant induced rheumatoid arthritis. Experimental groups were divided into 6 groups ; Normal, Control, $ST_{36}$, $SP_9$, $ST_{36}+SP_9$ and Non-Acupoint. Normal group, non-arthritic group, was injected normal saline, and the other groups were injected FCA. Each acupoint groups were treated by 2Hz electroacupuncture at each acupoints and NA group was treated by 2Hz electroacupuncture at non-acupoint. Each groups were evaluated by the number of NADPH-d positive neurons in primary somatosensory area(S1), secondary somatosensory area(S2), motor area and caudate putamen by using an image analyzer and a microscope. Results : 1. In S1, the number of NADPH-d positive neuron cells in the $ST_{36}$ group were significantly(p<0.05) increased compared with the control group. 2. In S2, the number of NADPH-d positive neuron cells in all electroacupuncture groups were not significantly changed compared with the control group. 3. In motor area, the number of NADPH-d positive neuron cells in $ST_{36}$ group, $SP_9$ group, NA group were significantly(p<0.05) increased compared with the control group. 4. In Caudate putamen, the number NADPH-d positive neuron cells in all electroacupuncture groups were significantly(p<0.05) decreased compared with the control group. Conclusions : Our result demonstrated that low frequency electroacupuncture on $ST_{36}$ & $SP_9$ normalized expression of NADPH-d positive neurons in the brain cortex of the rheumatoid arthritis model in rats.

• Journal of Acupuncture Research
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• v.24 no.2
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• pp.187-201
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• 2007

Objectives: This study was to investigate the effect of high frequency electro-acupuncture (EA) at $ST_{36}$ acupoint on the Freund's Complete Adjuvant (FCA)-induced arthritis in rats. Methods : Arthritis was induced by intradermal injection of FCA into base of tail. Experimental groups were divided into 4 groups; Normal, Control, $ST_{36}$ and Non-Acupoint (NA). $ST_{36}$ group was treated by 120Hz EA at $ST_{36}$ acupoint. Body weight, paw edema volume and ankle joint thickness were measured after treatment. And we investigated the effects of 120Hz EA via WBC count, segmen neurophil, lymphocyte, PGE2 assay and NADPH-d histochemistry. Results and Conclusions : The mean of body weight and ankle joint thickness of $ST_{36}$ group was increased compared with control group. The mean of paw edema volume, WBC count, segment neutrophil and lymphocyte of were not decreased with control group. The mean of PGE2 concentration ($1,851.00{\pm}160.11$) and NADPH-d positive neurons ($13.12{\pm}1.23$) were changed significantly (p

• Journal of Acupuncture Research
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• v.23 no.6
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• pp.207-219
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• 2006

Objectives : This study was to investigate the effect of low frequency electro-acupuncture (EA) at ST36 acupoint on the Freund's Complete Adjuvant(FCA)-induced arthritis in rats. Methods : Arthritis was induced by intradermal injection of FCA into base of tail. Experimental groups were divided into 4 groups; Normal, Control, ST36 and Non-Acupoint(NA). ST36 group was treated by 2 Hz EA at ST36 acupoint. Body weight, paw edema volun1e and ankle joint thickness were measured after treatment. And we investigated the effects of 2 Hz EA via WBC count, segment neurophil, lymphocyte, PGE2 assay and NADPH-d histochemistry. Results and Conclusion : The mean of body weight and ankle joint thickness of ST36 group was increased compared with control group. The mean of paw edema volume, WBC count, segment neutrophil and lymphocyte of were not decreased with control group. The mean of PGE2 concentration and NADPH-d positive neurons were changed significantly compared with control group.

• Journal of Acupuncture Research
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• v.20 no.5
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• pp.118-132
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• 2003

Objective: The aim of this study was to investigate the effects of Joksamni(ST36) combination on NAD PH-diaphorase, neuronal nitric oxide synthase(nNOS), neuropeptide Y(NPY) and vasoactive intestinal peptide (VIP) in the cerebral cortex of spontaneously hypertensive rat. Methods: The experimental groups were divided into four groups: Normal, Joksamni(ST36), Joksamni(ST36)+Eumneungcheon(SP9), and Joksamni(ST36)+Gokji(LI11). Needles were inserted into acupoints at the depth of 0.5cm with basic insertion method. Electroacupuncture was done under the condition of 2Hz electrical biphasic pulses with continuous rectangular wave lasting for 0.2ms until the muscles produced visible contractions. Such stimulation was applied continuously for 10 minutes, 1 time every 2 days for 10 sessions of treatments. Thereafter we evaluated changes in NADPH-d positive neurons histochemically and changes in nNOS, NPY and VIP positive neurons immunohistochemically. Results: The optical densities of NADPH-d positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group in all areas of cerebral cortex and Joksamni(ST36)+Gokji(LI11) group in primary somatosensory cortex, visual cortex, auditory cortex, perirhinal cortex were significantly increased as compared to the Joksamni(ST36) group. The optical densities of NADPH-d positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group with the exception of primary somatosensory cortex. The optical densities of nNOS positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group in all areas of cerebral cortex and Joksamni(ST36)+Gokji(LI11) group in auditory cortex, perirhinal cortex, insular cortex were significantly increased as compared to the Joksamni(ST36) group. The optical densities of nNOS positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in all areas of cerebral cortex as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group. The optical densities of NPY positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in primary motor cortex, primary somatosensory cortex, cingulate cortex as compared to the Joksamni (ST36) and Joksamni(ST36)+Eumneungcheon(SP9) groups. The optical densities of VIP positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group were significantly increased in all areas of cerebral cortex except for cingulate cortex as compared to the Joksamni(ST36) group. The optical densities of VIP positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in auditory cortex, cingulate cortex, perirhinal cortex as compared to the Joksamni(ST36) group. The optical densities of VIP positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in all areas of cerebral cortex as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group. Conclusions: The result demonstrated that electroacupuncture on Joksamni(ST36) and its combination change the activities of the NO system and peptidergic system in the cerebral cortex of SHR and that acupoint combination is one of the important parameters for the effects.

• Journal of Acupuncture Research
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• v.21 no.2
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• pp.205-222
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• 2004

Objective: To investigate the effects of acupointed Choksamni(ST36), Kokchi(LI11) and Arbitrary acupoint on NADPH-diaphorase, neuronal nitric oxide synthase(nNOS), neuropeptide Y(NPY) and vasoactive intestinal peptide(VIP) in the cerebral cortex of spontaneously hypertensive rats. Methods: The experimental groups were divided into four groups: Normal, Choksamni(ST36), Kokchi(LI11), arbitrary group. Needles were inserted into acupoints at the depth of 0.5 cm with basic insertion method. Such stimulation was applied continuously for 10 minutes, every other day, for 10 sessions of treatments. Thereafter we evaluated changes in NADPH-d-positive neurons histochemically and changes in nNOS, NPY and VIP-positive neurons immunohistochemically. Results : The optical densities of NADPH-d-positive neurons of all the Choksamni & Kokchi groups were significantly different in all areas of cerebral cortex as compared to arbitrary group. In motor1, sensory2, cingulate2, insular, peripheral, visual cortex there was a significant difference between Choksamni & Kokchi group. The optical densities of nNOS-positive neurons of Choksamni group were significantly different in all areas except for auditory, visual and pisiform cortex and Kokchi group in all areas except for auditory and pisiform cortex as compared to arbitrary group. And there was a significant difference in cingulate1, cingulate2, ectohinal, visual cortex between Choksamni & Kokchi group. The optical densities of NPY neurons of Choksamni group were significantly different in cingulate2, insular, pisiform cortex and Kokchi group in motor1, motor2, sensory1, cingulate2, ectorhinal cortex as compared to arbitrary group. And there was no significant difference between Choksamni & Kokchi group. The optical densities of VIP neurons of Choksamni group were significantly different in all areas except for motor1, auditory cortex and Kokchi group in sensory1, insular, ectorhinal, perirhinal, visual, pisiform cortex as compared to arbitrary group. And there was a significant difference in cingulate1, cingulate2, retrosplenial, auditory corterx between Choksamni & Kokchi group. Conclusion : Our results demonstrated that acupuncture on Choksamni(ST36) & Kokchi(LI11) changes the control activities of the NO system in the cerebral cortex of SHR and according to areas there were significant difference between two groups. In all cerebral cortex areas there were distributed NPY & VIP and there were no significant difference among Choksamni(ST36), Kokchi(LI11) and arbitrary group.

• Journal of Acupuncture Research
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• v.26 no.6
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• pp.51-65
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• 2009

Objectives : This study was built to investigate the effect of Trachelospermi Caulis herbal-acupuncture on the Collagen-induced arthritis(CIA) in rats. Methods : Arthritis was induced by intradermal injection of Bovine type II collagen solution into base of tail. Experimental group were divided into 5 groups ; Normal(N) group, Control(C) group, Trachelospermi Caulis high(TH) group, Trachelospermi Caulis low(TL) group, Saline(S) group(n=7 for each group). Normal group was had no management. Control group was injected with Bovine type II collagen solution and taken no treatment. Trachelospermi Caulis high group was injected with Bovine type II collagen solution and taken high-intensity(10mg/kg) herbal-acupuncture treatment on $ST_{36}$. Trachelospermi Caulis low group was injected with Bovine type II collagen solution and taken low-intensity(5mg/kg) herbal-acupuncture treatment on $ST_{36}$. Saline group was injected with Bovine type II collagen solution and taken saline injection on $ST_{36}$. Body weight, paw edema volume and ankle joint thickness were measured during experimental day. On the last experimental day, we analyzed WBC count, TNF-$\alpha$ & IL-$1{\beta}$ concentration, c-fos immunohistochemistry and NADPH-d histochemistry for evaluating the effect of Trachelospermi Caulis herbal-acupuncture. Results : The results were as follows ; 1. In the change of paw edema volume, TH group only has significant difference compared with C group. 2. In the change of ankle joint thickness, TH group only has significant difference compared with C group. 3. In WBC count of serum, TH, TL groups have significant decrease compared with C group. 4. In TNF-$\alpha$ concentration of effusion, TH, TL groups have significant decrease compared with C group. and TH group has significant decrease compared with TL group. 5. In IL-$1{\beta}$ concentration of effusion, TH, TL groups have significant decrease compared with C group. and TH group has significant decrease compared with TL group. 6. In c-fos positive neurons of S1S2(cortex) region, TH, TL, S groups have significant decrease compared with C group. 7. In NADPH-d positive neurons of CPu(caudate putamen) region, TH, TL groups have significant decrease compared with C group. 8. In NADPH-d positive neurons of Tfp(transverse fibers of pons) region, TH, TL, S groups have significant decrease compared with C group. and TH group has significant decrease compared with S group. Conclusions : According to above results, we hope that Trachelospermi Caulis herbal-acupuncture may have the effect that decreases progression and development of CIA. And it can be suggested that Trachelospermi Caulis herbal-acupuncture may reduce the expression of c-fos and NOS.

• Journal of Acupuncture Research
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• v.20 no.6
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• pp.148-159
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• 2003

Objective: This study examines the effects of acupoint combination on NADPH-diaphorase and neuronal nitric oxide synthase(nNOS) in the brainstem of spontaneously hypertensive rats. Methods: The changes of NADPH-d-positive neurons using a histochemical method and the changes of nNOS-positive neurons using an immunohistochemical method were evaluated. The optical densities of NADPH-d-positive neurons and nNOS-positive neurons of the Choksamni(ST36) Umnungchon(SP9) groups were significantly increased in all brainstem areas as compared to the Choksamni and Choksamni Kokchi(LI11) groups and decreased, with the exeption of the nNOS-positive neurons in the superficial gray of superior colliculus, as compared to the normal group. Results: Our results demonstrated that electroacupuncture changes the activity in the NO system in the brainstem of SHR and the acupoint combination is one of the important parameters for this effect.

• Korean Journal of Acupuncture
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• v.24 no.4
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• pp.221-231
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• 2007

목 적 : 본 연구의 목적은 시상하부에서 침처치에 대한 nitric oxide synthase (NOS)발현을 nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d)를 이용한 조직화학 염색법으로 관찰하였다. 실험방법 : 동물은 Balb/c (wild type) 와 Stat 4 knockout (KO) 생쥐를 사용하였다. 염증유도는 1% carrageenan 용액 (20ul/마리)을 발 뒤꿈치 표피에 주사하였고, 침 처치는 족삼리 (ST36)에 시침하였다. 침 처치 후 5시간까지 부종율을 부종측정기로 측정하였으며, 마지막으로 부종을 측정한 후 동물을 희생하여 뇌를 적출하여 고정하였다. 침에 대한 효과를 확인하기 위하여 NADPH-d 반응의 조직염색을 실시하였다. 염증유도와 그룹간의 유의성 검증은 one-way ANOVA를 사용하였다. 결 과 : 대조군인 Balb/c와 실험군인 stat4 KO 생쥐를 carrageenan으로 염증을 유도시에 대조군은 90%이상 유도된 반면, Stat4 KO 그룹은 50% 정도의 염증만이 유도되었다. 염증을 유도한 생쥐의 족삼리에 침 처치시 대조군은 1시간에서 약 40%정도 감소하였고 (P<0.05), Stat4 KO 실험군은 유의한 염증 감소율을 보이지 않았다. 시상하부의 lateral hypothalamic area (LHA)와 paraventricular nucleus (PVN)부위의 침에 대한 효과를 NADPH-d 에 양성으로 반응하는 세포수로 비교하여 다음과 같은 결과를 얻었다. (1) 대조군에서 염증 유도시 시상하부의 PVN는 NADPH-d 양성세포수가 감소하였고, LHA에서는 증가하였다. (2) 염증을 유도한 대조군에 침을 처치시 PVN은 세포수가 증가하였고, LHA에서는 감소하는 경향을 보였다. (3) 염증을 유도한 Stat4 KO 군에서는 시상하부의 PVN과 LHA부위 모두에서 NADPH-d 양성세포수가 감소하였고, 염증유도그룹에 침을 처치시 PVN과 LHA부위 모두에서 세포수가 증가함을 관찰 할 수 있었다. (4) 대조군과 실험군 모두에 salicylic acid로 비교하였더니 염증유도 효과 및 NADPH-d 세포 수에서 침 처치와 비슷한 결과를 나타내었다. 결 론 : 침은 염증을 유도한 생쥐에서 염증 감소에 유의한 효과가 있다. 염증을 유도한 Balb/c 와 Stat4 KO 생쥐에 침을 처치 시 시상하부의 NADPH-d 발현이 LHA부위와 PVN에서 서로 다르게 나타나는 것으로 나타난다. 이러한 현상은 침 효과가 시상하부의 위치에 대한 작용이 다르기 때문이라고 생각된다.

• Journal of Acupuncture Research
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• v.22 no.2
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• pp.13-18
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• 2005

This study was undertaken to investigate the effects of electroacupuncture(EA) on Choksamni(ST36), a well-known acupuncture site, on nicotinamide adenine dinucleotide phosphate-diaphorase(NADPH-d), neuropeptide Y(NPY) and vasoactive intestinal peptide(VIP) in the cerebral cortex of spontaneously hypertensive rats(SHR). EA on Choksamni was applied using 2Hz electrical biphasic pulses of 10min, 3 times a week for a total of 10 sessions. Thereafter we evaluated changes in NADPH-d-positive neurons histochemically and changes in NPY and VIP-positive neurons immunohistochemically. The optical density of NADPH-d-positive neurons in the Choksamni group was significantly lower in all areas of the cerebral cortex than in the control group. However, the optical density of NPY-positive neurons in the Choksamni group was similar to that of the controls in most areas of the cerebral cortex, with the exception of the primary motor and visual cortices. The optical density of VIP-positive neurons in the Choksamni group was significantly decreased as compared to the control group in most areas of the cerebral cortex, with the exception of the cingulate cortex. The present results demonstrated that EA on Choksamni changes the activity of the NO system, and that stimulation at the same level, causes selective changes within the peptidergic system in the cerebral cortex of SHR.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.156-162
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• 2012

A mannitol dehydrogenase (StMDH) gene was cloned from Salmonella typhimurium LT2 (KCTC 2421) and overexpressed in Escherichia coli. It has a 1,467 bp open reading frame encoding 488 amino acids with deduced molecular mass of 54 kDa, which shares approximately 36% of amino acid identity with known long-chain dehydrogenase/reductatse (LDR) family enzymes. The recombinant StMDH showed the highest activity at $30^{\circ}C$, and pH 5.0 and 10.0 for D-fructose reduction and D-mannitol oxidation, respectively. On the contrary, it has no activity on glucose, galactose, xylose, and arabinose. StMDH can catalyze the oxidative/reductive reactions between D-fructose and D-mannitol only in the presence of $NAD^+$/NADH as coenzymes. These results indicate that StMDH is a typical $NAD^+$/NADH-dependent mannitol dehydrogenase (E.C. 1.1.1.67).