Bloodletting Acupuncture at Jing-Well Points Alleviates Myocardial Injury in Acute Altitude Hypoxic Rats by Activating HIF-1α/BNIP3 Signaling-Mediated Mitochondrial Autophagy and Decreasing Oxidative Stress / 中国结合医学杂志

OBJECTIVE@#To explore the protective effect and possible mechanisms of bloodletting acupuncture at Jing-well points (BAJP) pre-treatment on acute hypobaric hypoxia (AHH)-induced myocardium injury rat.@*METHODS@#Seventy-five rats were randomly divided into 5 groups by a random number table: a control group (n=15), a model group (n=15), a BAJP group (n=15), a BAJP+3-methyladenine (3-MA) group (n=15), and a BANA (bloodletting at nonacupoint; tail bleeding, n=15) group. Except for the control group, the AHH rat model was established in the other groups, and the corresponding treatment methods were adopted. Enzyme-linked immunosorbent assay (ELISA) was used to detect creatine kinase isoenzyme MB (CK-MB) and cardiac troponins I (CTnI) levels in serum and superoxide dismutase (SOD) and malondialdehyde (MDA) levels in myocardial tissue. Hematoxylin-eosin (HE) staining was used to observe myocardial injury, and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was used to observe cell apoptosis. Transmission electron microscopy detection was used to observe mitochondrial damage and autophagosomes in the myocardium. The mitochondrial membrane potential of the myocardium was analyzed with the fluorescent dye JC-1. Mitochondrial respiratory chain complex (complex I, III, and IV) activities and ATPase in the myocardium were detected by mitochondrial respiratory chain complex assay kits. Western blot analysis was used to detect the autophagy index and hypoxia inducible factor-1α (HIF-1α)/Bcl-2 and adenovirus E1B 19k Da-interacting protein 3 (BNIP3) signaling.@*RESULTS@#BAJP reduced myocardial injury and inhibited myocardial cell apoptosis in AHH rats. BAJP pretreatment decreased MDA levels and increased SOD levels in AHH rats (all P<0.01). Moreover, BAJP pretreatment increased the mitochondrial membrane potential (P<0.01), mitochondrial respiratory chain complex (complexes I, III, and IV) activities (P<0.01), and mitochondrial ATPase activity in AHH rats (P<0.05). The results from electron microscopy demonstrated that BAJP pretreatment improved mitochondrial swelling and increased the autophagosome number in the myocardium of AHH rats. In addition, BAJP pretreatment activated the HIF-1α/BNIP3 pathway and autophagy. Finally, the results of using 3-MA to inhibit autophagy in BAJP-treated AHH rats showed that suppression of autophagy attenuated the treatment effects of BAJP in AHH rats, further proving that autophagy constitutes a potential target for BAJP treatment of AHH.@*CONCLUSION@#BAJP is an effective treatment for AHH-induced myocardial injury, and the mechanism might involve increasing HIF-1α/BNIP3 signaling-mediated autophagy and decreasing oxidative stress.

Bloodletting Acupuncture at Jing-Well Points on Hand Induced Autophagy to Alleviate Brain Injury in Acute Altitude Hypoxic Rats by Activating PINK1/Parkin Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the protective effect of bloodletting acupuncture at twelve Jing-well points on hand (BAJP) on acute hypobaric hypoxia (AHH)-induced brain injury in rats and its possible mechanisms.@*METHODS@#Seventy-five Sprague Dawley rats were divided into 5 groups by a random number table (n=15), including control, model, BAJP, BAJP+3-methyladenine (3-MA), and bloodletting acupuncture at non-acupoint (BANA, tail tip blooding) groups. After 7-day pre-treatment, AHH models were established using hypobaric oxygen chambers. The levels of S100B, glial fibrillary acidic protein (GFAP), superoxide dismutase (SOD), and malondialdehyde (MDA) in serum were measured by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method were used to assess hippocampal histopathology and apoptosis. Transmission electron microscopy assay was used to observe mitochondrial damage and autophagosomes in hippocampal tissues. Flow cytometry was used to detect mitochondrial membrane potential (MMP). The mitochondrial respiratory chain complexes I, III and IV activities and ATPase in hippocampal tissue were evaluated, respectively. Western blot analysis was used to detect the protein expressions of Beclin1, autophagy protein 5 (ATG5), microtubule-associated protein 1 light chain 3 beta (LC3B), phosphatase and tensin homolog induced kinase 1 (PINK1), and Parkin in hippocampal tissues. The mRNA expressions of Beclin1, ATG5 and LC3-II were analyzed by quantitative real-time polymerase chain reaction.@*RESULTS@#BAJP treatment reduced hippocampal tissue injury and inhibited hippocampal cell apoptosis in AHH rats. BAJP reduced oxidative stress by decreasing S100B, GFAP and MDA levels and increasing SOD level in the serum of AHH rats (P<0.05 or P<0.01). Then, BAJP increased MMP, the mitochondrial respiratory chain complexes I, III and IV activities, and the mitochondrial ATPase activity in AHH rats (all P<0.01). BAJP improved mitochondrial swelling and increased the autophagosome number in hippocampal tissue of AHH rats. Moreover, BAJP treatment increased the protein and mRNA expressions of Beclin1 and ATG5 and LC3-II/LC3-I ratio in AHH rats (all P<0.01) and activated the PINK1/Parkin pathway (P<0.01). Finally, 3-MA attenuated the therapeutic effect of BAJP on AHH rats (P<0.05 or P<0.01).@*CONCLUSION@#BAJP was an effective treatment for AHH-induced brain injury, and the mechanism might be through reducing hippocampal tissue injury via increasing the PINK1/Parkin pathway and enhancement of mitochondrial autophagy.

Bloodletting Puncture at Hand Twelve Jing-Well Points Relieves Brain Edema after Severe Traumatic Brain Injury in Rats via Inhibiting MAPK Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate whether blood-brain barrier (BBB) served a key role in the edema-relief effect of bloodletting puncture at hand twelve Jing-well points (HTWP) in traumatic brain injury (TBI) and the potential molecular signaling pathways.@*METHODS@#Adult male Sprague-Dawley rats were assigned to the sham-operated (sham), TBI, and bloodletting puncture (bloodletting) groups (n=24 per group) using a randomized number table. The TBI model rats were induced by cortical contusion and then bloodletting puncture were performed at HTWP twice a day for 2 days. The neurological function and cerebral edema were evaluated by modified neurological severity score (mNSS), cerebral water content, magnetic resonance imaging and hematoxylin and eosin staining. Cerebral blood flow was measured by laser speckles. The protein levels of aquaporin 4 (AQP4), matrix metalloproteinases 9 (MMP9) and mitogen-activated protein kinase pathway (MAPK) signaling were detected by immunofluorescence staining and Western blot.@*RESULTS@#Compared with TBI group, bloodletting puncture improved neurological function at 24 and 48 h, alleviated cerebral edema at 48 h, and reduced the permeability of BBB induced by TBI (all P<0.05). The AQP4 and MMP9 which would disrupt the integrity of BBB were downregulated by bloodletting puncture (P<0.05 or P<0.01). In addition, the extracellular signal-regulated kinase (ERK) and p38 signaling pathways were inhibited by bloodletting puncture (P<0.05).@*CONCLUSIONS@#Bloodletting puncture at HTWP might play a significant role in protecting BBB through regulating the expressions of MMP9 and AQP4 as well as corresponding regulatory upstream ERK and p38 signaling pathways. Therefore, bloodletting puncture at HTWP may be a promising therapeutic strategy for TBI-induced cerebral edema.

Origin and relationship of -well points and Shixuan (EX-UE 11) / 中国针灸

Both the twelve -well points and Shixuan (EX-UE 11) are the commonly used first-aid points. These two kinds of acupoints are located closely and similar in function, hence, they are often confused in application. In order to explore the origin of their location and theory as well as their relationship, the relevant data were retrieved. It has been found that the relationship between -well points and Shixuan (EX-UE 11) is traced at the earliest time to (). It is believed that the -well points refer to the starting points or the ending points of the twelve regular meridians and Shixuan (EX-UE 11) are located at the crossing sites of - related meridians of the regular meridians. These two kinds of acupoints are interconnected, share the same source and are also different from each other. Qiduan (EX-LE 12) is also named as foot-Shixuan. Shixuan (EX-UE 11) and Qiduan (EX-LE 12) can be regarded as the same category, just like -well points. In clinical practice, the -well points are generally selected in treatment of internal diseases, local diseases and those on the running course of meridians. They can be used separately in treatment. Shixuan (EX-UE 11) is the first option, or combined with Qiduan (EX-LE 12) in the emergent treatment of tense syndrome and syncope. The -well points and Shixuan (EX-UE 11) are different even though sharing the same origin. They are mutually benefited and supplemented with each other in clinical practice.

Bloodletting acupuncture at -well points along three- meridians of foot combined with acupuncture on migraine:a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the analgesic effect of bloodletting acupuncture at -well points along three- meridians of foot combined with routine acupuncture and simple routine acupuncture on migraine.@*METHODS@#A total of 60 patients with migraine were randomized into an observation group and a control group, 30 cases in each one, of which, 4 cases were dropped out in the observation group, 1 case was dropped out in the control group. In the observation group, bloodletting acupuncture at -well points combined with routine acupuncture were applied. The bloodletting acupuncture was applied at corresponding -well points of three- meridians of foot [Lidui (ST 45), Zhiyin (BL 67), Zuqiaoyin (GB 44)] according to pain location. And routine acupuncture was adopted at Sizhukong (TE 23), Shuaigu (GB 8), Taiyang (EX-HN 5), Fengchi (GB 20), Hegu (LI 4), Taichong (LR 3), Zulinqi (GB 41), Yanglingquan (GB 34) and Waiguan (TE 5). In the control group, routine acupuncture was applied, acupoint selection and operation were the same as the observation group. The treatment was given once a day, 30 min a time, 5 days as one course with 2 days interval, and 2 courses were required. Before treatment, immediately after needle withdrawal, 4 h after needle withdrawal and after 2 courses of treatment, the visual analogue scale (VAS) score was compared in the two groups. Before and after treatment, the migraine comprehensive score was observed in the two groups, and the therapeutic effect was evaluated.@*RESULTS@#Immediately after needle withdrawal, 4 h after needle withdrawal and after 2 courses of treatment, the VAS scores in the two groups were decreased (0.05).@*CONCLUSION@#Bloodletting acupuncture at -well points along three- meridians of foot combined with routine acupuncture and simple routine acupuncture have analgesic effect, and the combined therapy is superior to simple routine acupuncture.

Research advances in the clinical effect of bloodletting puncture at well-points in treatment of acute central nervous injury / 针刺研究

Bloodletting puncture at twelve well-points is a characteristic emergency therapy in traditional Chinese medicine. This article reviewed the research advances in the clinical effect of this therapy in the treatment of acute central nervous injury and its mechanism of action over the past 30 years, and it is found that this therapy can effectively improve disturbance of consciousness, neurological defects, and cerebral edema caused by stroke, traumatic brain injury, and carbon monoxide poisoning. The mechanism involves the improvement of cerebral blood flow and tissue oxygen supply, repair of the blood-brain barrier, and regulation of local ion balance. Well-designed clinical trials and in-depth research on biological mechanisms should be performed in future to promote and guide its clinical application.

Effect of bloodletting acupuncture at twelve -well points of hand on microcirculatory disturbance in mice with traumatic brain injury / 中国针灸

OBJECTIVE@#To observe the effect of bloodletting acupuncture at twelve -well points of hand on microcirculatory disturbance in mice with traumatic brain injury (TBI), and to explore the protective effect of bloodletting therapy on TBI.@*METHODS@#Sixty clean adult male C57BL/6J mice were randomly divided into a sham-operation group, a model group and a treatment group, 20 mice in each group. The TBI model was established by using electronic controlled cerebral cortex impact instrument in the model group and the treatment group. The mice in the treatment group were treated with bloodletting acupuncture at bilateral "Shaoshang" (LU 11), "Shangyang" (LI 1), "Zhongchong" (PC 9), "Guanchong" (TE 1), "Shaochong" (HT 9) and "Shaoze" (SI 1) immediately after trauma. The mice in the sham-operation group only opened the bone window but did not receive the strike. The regional cerebral blood flow (rCBF) was monitored by laser speckle contrast analysis (LASCA) using a PeriCam PSI System before trauma, immediately after trauma and 1, 2, 12, 24, 48, 72 h after trauma. The brain water content was measured by wet-dry weight method 24 h after trauma. The severity of functional impairment at 2, 12, 24, 48 and 72 h after trauma was evaluated by modified neurological scale scores (mNSS).@*RESULTS@#① 2 h after trauma, the mNSS in the model group and treatment group were >7 points, suggesting the successful establishment of model; compared with the sham-operation group, the mNSS was increased significantly from 12 to 72 h after trauma in the model group ( all <0.01), but the mNSS in the treatment group was significantly lower than that in the model group from 2 to 24 h after trauma (<0.01, <0.05). ② Compared with the sham-operation group, rCBF in the model group was decreased significantly immediately after trauma (<0.01), and the rCBF in the model group was lower than that in the sham-operation group from 1 to 72 h after trauma ( all <0.01); rCBF in the treatment group began to rise and was significantly higher than that in the model group 1-2 h after trauma (<0.01); 12-48 h after trauma, the increasing of rCBF in the two groups tended to be gentle until 72 h after injury, and rCBF in the model group was decreased while that in the treatment group continued to rise and was higher than that in the model group (<0.01). ③ 24 h after trauma, the brain water content in the model group was significantly higher than that in the sham-operation group (<0.01), and brain water content in the treatment group was significantly lower than that in the model group (<0.01).@*CONCLUSION@#The bloodletting acupuncture at twelve -well points of hand could improve microcirculation disturbance, increase microcirculation perfusion, alleviate secondary brain edema and promote the recovery of nerve function in mice with TBI.

Clinical study on post-stroke urinary retention treated with acupuncture at the twelve-well points and bladder function training / 中国针灸

<p><b>OBJECTIVE</b>To explore the methods for the therapeutic effect improvement in the treatment of post-stroke urinary retention.</p><p><b>METHODS</b>Sixty-three patients of post-stroke urinary retention were randomized into an observation group (32 cases) and a control group (31 cases). The routine clinical medication of neurology and basic rehabilitation were adopted in the two groups. Additionally, in the control group, the intermittent urinary catheterization and bladder function training were applied. The duration and frequency of catheterization were determined by the autonomic urination and residual urine volume every day. In the observation group, on the basis of the treatment as the control group, acupuncture was applied to the twelve-well points in the sequence offlowing among the twelve meridians [Shaoshang (LU 11), Shangyang (LI 1), Lidui (ST 45), Yinbai (SP 1), Shaochong (HT 9), Shaoze (SI 1), Zhiyin (BL 67), Yongquan (KI 1), Zhongchong (PC 9), Guanchong (TE 1), Zuqiaoyin (GB 44) and Dadun (LR 1)]. Acupuncture was given once a day, 20 treatments were required. In 20 treatments, the clinical therapeutic effects and the residual urine volume were observed.</p><p><b>RESULTS</b>The total effective rate was 90.6% (29/32) in the observation group, better than 67.7% (21/31) in the control group (<0.01)). After treatment, the residue urine volume was all reduced apparently in the patients of the two groups (both<0.01). The result of the residue urine volume in the observation group was lower apparently than that in the control group (<0.01).</p><p><b>CONCLUSION</b>Acupuncture at the-well points in the sequence offlowing among meridians combined with bladder function training achieve the apparent therapeutic effects on post-stroke urinary retention. The results are better than those achieved by the routine western medicine with bladder function training involved.</p>

The Effect of Hand Twelve Jing-Well Points Bloodletting on Blood-brain Barrier Permeability in PMCAO Rats / 上海针灸杂志

Objective To investigate the effect of hand twelve Jing-Well points bloodletting on blood-brain barrier (BBB) permeability in the early stage of focal cerebral ischemic injury in rats with permanent middle cerebral artery occlusion (pMCAO). Method Male Wister rats were randomized into sham operation, model and bloodletting groups. A model of middle cerebral artery occlusion was made by Longa's method. 2%Evans blue (EB) was injected into the caudal vein at three time points:5, 24 and 72 hrs after model making. EB exudation amount was assessed by measurement of EB absorbance in brain tissue to observe BBB permeability. Result Cerebral EB exudation amount was significantly higher in the model group of rats than in the sham operation group (P<0.01) and significantly lower in the bloodletting group than in the model group (P<0.01). Conclusion Hand twelve Jing-Well points bloodletting can reduce blood-brain barrier permeability in pMCAO rats. Its effect is highly significant at 24 and 72 hrs after ischemia.

Observations on the Efficacy of Jin’s Three Needles plus Twelve Jing-Well Points in Treating Post-stroke Shoulder-hand Syndrome / 上海针灸杂志

Objective To investigate the clinical efficacy of Jin’s Three Needles plus twelve Jing-Well points in treating post-stroke shoulder-hand syndrome. Methods Sixty patients meeting the inclusion criteria of post-stroke shoulder-hand syndrome were randomly allocated to treatment and control groups, 30 cases each. The treatment group received acupuncture with Jin’s Three Needles plus twelve Jing-Well points and the control group, conventional acupuncture. The Clinical Neurological Disfunction Scale (CNDS) score, the Fugl-Meyer Assessment (FMA) score, the Visual Analogue Scale (VAS) score and the Functiional Comprehensive Assessment (FCA) score were observed in the two groups before and after two weeks of treatment. The clinical therapeutic effects were compared between the two groups. Results There were statistically significant pre-/post-treatment differences in the CNDS, FMA, VAS and FCA scores in the two groups (P<0.05). The total efficacy rate and the cure and marked efficacy rate were 96.7%and 55.7%in the combination group, respectively, in the treatment group and 83.3%and 26.7%, respectively, in the control group;there were statistically significant differences between the two groups (P<0.05). Conclusion Acupuncture with Jin’s Three Needles plus twelve Jing-Well points is an effective way to treat post-stroke shoulder-hand syndrome.