Korean red ginseng promotes hippocampal neurogenesis in mice

Neurogenesis in the adult hippocampus plays a major role in cognitive ability of animals including learning and memory.Korean red ginseng (KRG) has long been known as a medicinal herb with the potential to improve learning and memory;however,the mechanisms are still elusive.Therefore,we evaluated whether KRG can promote cognitive function and enhance neurogenesis in the hippocampus.Eight-week-old male C57BL/6 mice received 50 mg/kg of 5-bromo-2′-deoxyuridine (BrdU) intraperitoneally and 100 mg/kg of KRG or vehicle orally once a day for 14 days.Pole,Rotarod and Morris water maze tests were performed and the brains were collected after the last behavioral test.Changes in the numbers of BrdU- and BrdU/ doublecortin (DCX;a marker for neuronal precursor cells and immature neurons)-positive cells in the dentate gyrus and the gene expression of proliferating cell nuclear antigen (a marker for cell differentiation),cerebral dopamine neurotrophic factor and ciliary neurotrophic factor in the hippocampus were then investigated.KRG-treated mice came down the pole significantly faster and stood on the rotarod longer than vehicle-treated mice.The Morris water maze test showed that KRG administration enhanced the learning and memory abilities significantly.KRG also significantly increased BrdU- and BrdU/DCX-positive cells in the dentate gyrus as well as the proliferating cell nuclear antigen,cerebral dopamine neurotrophic factor and ciliary neurotrophic factor mRNA expression levels in the hippocampus compared to vehicle.Administration of KRG promotes learning and memory abilities,possibly by enhancing hippocampal neurogenesis.This study was approved by the Pusan National University Institutional Animal Care and Use Committee (approval No.PNU-2016-1071) on January 19,2016.

Effects of electroacupuncture on the functionality of NG2-expressing cells in perilesional brain tissue of mice following ischemic stroke

Neural/glial antigen 2(NG2)-expressing cells has multipotent stem cell activity under cerebral ischemia.Our study examined the effects of electroacupuncture(EA)therapy(2 Hz,1 or 3 mA,20 minutes)at the Sishencong acupoint on motor function after ischemic insult in the brain by investigating the rehabilitative potential of NG2-derived cells in a mouse model of ischemic stroke.EA stimulation alleviated motor deficits caused by ischemic stroke,and 1 mA EA stimulation was more efficacious than 3 mA EA stimulation or positive control treatment with edaravone,a free radical scavenger.The properties of NG2-expressing cells were altered with 1 mA EA stimulation,enhancing their survival in perilesional brain tissue via reduction of tumor necrosis factor alpha expression.EA stimulation robustly activated signaling pathways related to proliferation and survival of NG2-expressing cells and increased the expression of neurotrophic factors such as brain-derived neurotrophic factor,tumor growth factor beta,and neurotrophin 3.In the perilesional striatum,EA stimulation greatly increased the number of NG2-expressing cells double-positive for oligodendrocyte,endothelial cell,and microglia/macrophage markers(CC1,CD31,and CD68).EA therapy also greatly activated brain-derived neurotrophic factor/tropomyosin receptor kinase B and glycogen synthase kinase 3 beta signaling.Our results indicate that EA therapy may prevent functional loss at the perilesional site by enhancing survival and differentiation of NG2-expressing cells via the activation of brain-derived neurotrophic factor-induced signaling,subsequently ameliorating motor dysfunction.The animal experiments were approved by the Animal Ethics Committee of Pusan National University(approval Nos.PNU2019-2199 and PNU2019-2884)on April 8,2019 and June 19,2019.

Analysis of Herbal Medicine Prescriptions for Patients in An Academic Korean Medical Hospital:A Cross Sectional Study of Electronic Medical Records(2010–2013)

Objective： To obtain fundamental information for the standardization of herbal medicine in Korea. Methods： We analyzed the herbal medicine prescription data of patients at the Pusan National University Korean Medicine Hospital from March 2010 to February 2013. We used the Dongui-Bogam （Dong Yi Bao Jian） to classify prescribed herbal medicines. Results： The study revealed that the most frequently prescribed herbal medicine was ‘Liuwei Dihuang Pill （LWDHP, 六味地黄丸）＇ which was used for invigorating ‘Shen （Kidndy）-yin＇. ‘LWDHP＇ was most frequently prescribed to male patients aged 50-59, 60-69, 70-79 and 80-89 years, and ‘Xionggui Tiaoxue Decoction （XGTXD, 芎归调血饮）＇ was most frequently prescribed to female patients aged 30-39 and 40-49 years. According to the International Classification of Diseases （ICD） codes,‘Diseases of the musculoskeletal system and connective tissue＇ showed the highest prevalence. ‘LWDHP＇ and ＇XGTXD＇ was the most frequently prescribed in categories 5 and 3, respectively. Based on the percentage of prescriptions for each sex, ‘Ziyin Jianghuo Decoction （滋阴降火汤）＇ was prescribed to mainly male patients, and ‘XGTXD＇ with ‘Guima Geban Decoction （桂麻各半汤）＇ were prescribed to mainly female patients. Conclusion： This study analysis successfully determined the frequency of a variety of herbal medicines, and many restorative herbal medicines were identified and frequently administered.

Muscovite nanoparticles mitigate neuropathic pain by modulating the inflammatory response and neuroglial activation in the spinal cord

Despite numerous efforts to overcome neuropathic pain,various pharmacological drugs often fail to meet the needs and have many side effects.Muscovite is an aluminosilicate mineral that has been reported to have an anti-inflammatory effect,but the efficacy of muscovite for neuropathic pain has not been investigated.Here,we assessed whether muscovite nanoparticles can reduce the symptoms of pain by controlling the inflammatory process observed in neuropathic pain.The analgesic effects of muscovite nanoparticles were explored using partial sciatic nerve ligation model of neuropathic pain,in which one-third to onehalf of the nerve trifurcation of the sciatic nerve was tightly tied to the dorsal side.Muscovite nanoparticles(4 mg/100μL)was given intramuscularly to evaluate its effects on neuropathic pain(3 days per week for 4 weeks).The results showed that the muscovite nanoparticle injections significantly alleviated partial sciatic nerve ligation-induced mechanical and cold allodynia.In the spinal cord,the muscovite nanoparticle injections exhibited inhibitory effects on astrocyte and microglia activation and reduced the expression of pro-inflammatory cytokines,such as interleukin-1β,tumor necrosis factor-α,interleiukin-6 and monocyte chemoattractant protein-1,which were upregulated in the partial sciatic nerve ligation model.Moreover,the muscovite nanoparticle injections resulted in a decrease in activating transcription factor 3,a neuronal injury marker,in the sciatic nerve.These results suggest that the analgesic effects of muscovite nanoparticle on partial sciatic nerve ligation-induced neuropathic pain may result from inhibiting activation of astrocytes and microglia as well as pro-inflammatory cytokines.We propose that muscovite nanoparticle is a potential anti-nociceptive candidate for neuropathic pain.All experimental protocols in this study were approved by the Institutional Animal Ethics Committee(IACUC)at Dongguk University,South Korea(approval No.2017-022-1)on September 28,2017.

Korean red ginseng decreases 1-methyl-4-phenylpyridinium-induced mitophagy in SH-SY5Y cells

Objective:Mitophagy is known to contribute towards progression of Parkinson’s disease.Korean red ginseng(KRG)is a widely used medicinal herb in East Asia,and recent studies have reported that KRG prevents 1-methyl-4-phenylpyridinium ion(MPP^(+))-induced cell death.This study was undertaken to investigate whether KRG suppresses MPP^(+)-induced apoptosis and mitophagy.Methods:SH-SY5 Y cells were incubated with KRG for 24 h,and subsequently exposed to MPP^(+).The MPP^(+)-induced cell death was confirmed with the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay,and the terminal deoxynucleotidyl transferase-mediated d UTP nick end-labeling assay.Changes in the structure and function of mitochondria were confirmed using mitotracker,Mito SOX red mitochondrial superoxide indicator,parkin,and phosphatase and tensin homolog deleted on chromosome ten-induced putative kinase 1(PINK1)immunofluorescent staining.Western blotting was performed to evaluate the expression of apoptosis-related factors in whole cells,including Bax,Bcl-2 and cleaved caspase-3,and mitophagy-related factors in the mitochondrial fraction,including cytochrome c,parkin,PINK1,translocase of the outer membrane 20(TOM20),p62 and Beclin 1.Results:MPP^(+)induced cell death by cytochrome c release and caspase-3 activation;however,this effect was suppressed by KRG’s regulation of the expressions of Bcl-2 and Bax.Moreover,MPP^(+)exposure increased the mitochondrial expressions of parkin,PINK1,Beclin 1 and p62,and decreased TOM20,cytochrome c and Bcl-2 expressions.These MPP^(+)-induced changes in the mitochondrial fraction were attenuated by treatment with KRG.Conclusion:KRG effectively prevents MPP^(+)-induced SH-SY5 Y cell death by regulating cytochrome c release from mitochondria and PINK1/parkin-mediated mitophagy,through regulation of the Bcl-2 family.

Acupuncture Points for Treating Parkinson's Disease Based on Animal Studies

Parkinson＇s disease （PD） is a well-known neurodegenerative disease caused by dopaminergic cell death in the nigrostriatal pathway. Recent studies have shown that acupuncture can be a potential therapy for the treatment of PD, but it is not clear which acupuncture points （acupoints） play major roles in reliving symptoms of PD. Yanglingquan （GB 34）, Zusanli （ST 36）, Fengfu （GV 16）, Taichong （LR 3）, Baihui （GV 20） and Dazhui （GV 14） acupoints have frequently been to investigate the effectiveness and action mechanism of acupuncture for treating PD, but it is not clear why they were selected. This review summarizes the current understanding of the acupoints for PD treatment based on Oriental medicine theories and on the accumulated findings from previous animal studies. The results of this study will be useful to development of a strategy for future research in this field.

Subacute electroacupuncture at Baihui (GV 20) and Dazhui (GV 14) promotes post-stroke functional recovery via neurogenesis and astrogliosis in a photothrombotic stroke mouse model

OBJECTIVE:To investigate the optimal timing and underlying mechanism of electroacupuncture(EA)at Baihui(GV 20)and Dazhui(GV 14)for improved long-term functional recovery after focal cerebral ischemia in a photothrombotic stroke mouse model.METHODS:Totally 50 adult male C57 BL/6 J mice were assigned into 5 groups:(a)the control group,sham-operated mice(n=10);(b)the vehicle group,focal cerebral ischemia induction without EA(n=10);(c)the acute EA group,mice received EA immediately post-ischemia,followed by once-daily treatments for 7 consecutive days(n=10);(d)the subacute EA group,mice received EA 4 days post-ischemia,followed by once-daily treatments for 7 consecutive days(n=10);(e)the delayed EA group.EA stimulation(2 Hz,2 V for 20 min)was applied to acupuncture points(acupoints),Baihui(GV 20)and Dazhui(GV 14),once a day for 7 consecutive days beginning immediately(acute treatment),4 d(subacute treatment)and 10 d(delayed treatment)after focal cerebral ischemia in C57 BL/6 J mice.Behavioral assessments were conducted 21 and 28 d post-ischemia and histopathological analyses were performed 28 days post-ischemia.RESULTS:The subacute EA treatment at Baihui(GV20)and Dazhui(GV 14)significantly improved functional recovery compared to the vehicle group 28 d after ischemic brain injury,although brain atrophy was not reduced.The number of Neu N+and Neu N+/Brd U+cells as well as GFAP intensity in the ipsilateral cortex were significantly increased in the subacute group compared to the vehicle group 28 d post-ischemia.We concluded that EA stimulation 4 d post-ischemia(subacute treatment)enhanced neurogenesis and astrogliosis,likely contributing to long-term functional recovery following focal cerebral ischemia.CONCLUSION:Our findings suggest that the timing of the EA therapy at Baihui(GV 20)and Dazhui(GV 14)determines the therapeutic effects in mice with focal cerebral ischemia induced by photothrombotic occlusion.

Syndrome Pattern and Its Application in Parallel Randomized Controlled Trials

Syndrome pattern （SP） is a core concept of Chinese medicine （CM） and is used to diagnose and treat patients based on an overall analysis of symptoms and signs. This study aimed to systematically review randomized controlled trials （RCTs） using the SP concept and to demonstrate how the SP concept could be applied to the study design of parallel RCTs, considering a gold standard of clinical research. After conducting a brief systematic review by way of a PubMed search, we analyzed how the SP concept was applied to the design of RCT in a CM herbal medicine trial. We then formulated possible research questions, applied the SP concept to answer the research questions, and suggested possible RCT designs to be used for conducting future trials. Fourteen RCTs were included in our systematic review, and three key points of the SP concept were formulated for the design of parallel RCTs： the time point of SP diagnosis between before and after randomization; the relationship between the international classification of diseases （ICD） and SP for the inclusion of target population; and the proper diagnostic method of SP. In this study, we formulated three possible research questions and then suggested perspectives for five possible RCT models arrived at using SP concepts. Future trials applying SP concept to RCTs should overcome the shortcomings of past SP trials, moving CM forward from experience-based to evidence-based medicine.