Blood-letting punctures at twelve Jing-Well points of the hand can treat cerebral ischemia in a similar manner to mannitol

A rat model of middle cerebral artery permanent occlusion was established using the modified Longa method. Successfully established model animals were treated by blood-letting puncture at twelve Jing-Well points of the hand, and/or by injecting mannitol into the caudal vein twice daily. Brain tissue was collected at 24, 48 and 72 hours after modeling, and blood was collected through the retinal vein before Evans blue was injected, approximately 1 hour prior to harvesting of brain tissue. Results showed that Evans blue leakage into brain tissue and serum nitric oxide synthase activity were significantly increased in model rats. Treatment with blood-letting punctures at twelve Jing-Well points of the hand and/or injection of mannitol into the caudal vein reduced the amount of Evans blue leakage into the brain tissue and serum nitric oxide synthase activity to varying degrees. There was no significant difference between single treatment and combined treatment. Experimental findings indicate that blood-letting punctures at twelve Jing-Well points of the hand can decrease blood-brain barrier permeability and serum nitric oxide synthase activity in rats following middle cerebral artery occlusion, and its effect is similar to that of mannitol injection alone and Jing-Well points plus mannitol injection.

Principle and operation of Jing-well point temperatures' test

Principle and operation of Jing-well point temperatures' test is a method to diagnose and treat diseases by measuring and comparing the symmetrical Jing-well point temperature of human body 12 meridians.It is an important supplement to traditional Chinese medicine four diagnoses. The theoretical basis are that the Jing-well is closely related to Meridians and Viscera, the basic traditional Chinese medicine principle of 'governing exterior to infer interior' and the balancing phenomenon of Meridians. In this paper, the operation steps and treatment measures of Jing-well point temperatures' test are put forward, and 2 successful cases are given. At the end of this paper, the prospect of this method is discussed.

Interpretation of data and treatment of Jing-well point temperatures test

The Jing-well point temperatures test method is a method to diagnose and guide the treatment of diseases by measuring the subjects' symmetrical well point temperature. it is improved from the method of knowing heat sensitivity. The application of Jing-well point temperatures test method is wide, and it can be used in internal and external gynecology and pediatrics and facial features department. at the same time, it has the advantage of objective and accurate diagnosis. The old law has some shortcomings, such as poor intuition, unavoidable omission of information, incomplete interpretation of information and so on. In this paper, Excel software is used to transform the data into line chart form, which improves the intuition and comprehensiveness of this method, so that the data can be better interpreted and used. It is newly proposed in this article that in addition to observing the longitudinal di fference of well point temperature, more attention should be paid to the horizontal contrast difference of well point temperature in different meridians. The article also summarizes a number of treatment methods, including acupuncture, moxa moxibustion, cupping and scraping, and the selection of acupoints, including mother acupoints, tenderness points and heat-sensitive moxibustion, so that doctors can combine traditional Chinese medicine professional knowledge in clinic.

Bloodletting Acupuncture at Jing-Well Points Alleviates Myocardial Injury in Acute Altitude Hypoxic Rats by Activating HIF-1 α/BNIP3 Signaling-Mediated Mitochondrial Autophagyand Decreasing Oxidative Stress

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(CTn I) 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 d UTP-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/BNIP3pathway 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

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-Ⅰratio 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.

井穴放血减轻全身炎症反应防治大鼠心脏骤停后脑损伤

目的探讨十二井穴放血疗法对大鼠心脏骤停后脑损伤的保护作用及其机制。方法18只SD大鼠随机分配到假手术组、模型组和井穴组3组,每组6只。假手术组不诱导心脏骤停,模型组采用气管夹闭法诱导心脏骤停后进行心肺复苏,井穴组在模型组基础上加用手十二井穴刺络放血。观察不同时间点神经功能缺陷评分、血清IL-6、TNF-α水平,并对72 h后处死大鼠取大脑海马组织进行尼氏染色及TUNEL法检测细胞凋亡。结果复苏后模型组和井穴组神经功能缺陷评分均较假手术组显著下降(P<0.05),其中6 h最为明显,至72 h井穴组和假手术组比较,差异无统计学意义(P>0.05)。复苏后72 h尼氏染色显示假手术组大鼠海马神经元细胞细胞核大,核仁明显;与模型组比较,井穴组大鼠脑组织神经元水肿较轻,细胞核形态较好。复苏后72 h时后,与假手术组比较,模型组和井穴组凋亡指数显著增高(P<0.05);与模型组比较,井穴组凋亡指数显著降低(P<0.05)。复苏后6 h后与假手术比较,模型组和井穴组血清IL-6、TNF-α水平显著升高(P<0.05)。与模型组比较,井穴组血清IL-6于复苏后24 h、72 h显著降低(P<0.05),血清TNF-α于复苏后24 h、48 h及72 h显著降低(P<0.05)。结论减轻心脏骤停后机体炎症反应可能是十二井穴刺络放血防治复苏后脑损伤的机制之一。

Wake-Promoting Effect of Bloodletting Puncture at Hand Twelve Jing-Well Points in Acute Stroke Patients:A Multi-center Randomized Controlled Trial

Objective To assess the effect and safety of bloodletting puncture at hand twelve Jing-Well points(HTWPs)in acute stroke patients with conscious disturbance.Methods In this multi-center and randomized controlled trial,360 patients suffered from ischemic or hemorrhagic stroke with conscious disturbance within 48 h from the onset of symptom were divided into bloodletting(180 cases)and control(180 cases)groups using a block randomization.Patients in both groups received routine Western medicine,and patients in the bloodletting group received additional bloodletting puncture at HTWPs on admission immediately before conventional treatment.The primary outcome measure was Glasgow Coma Scale(GCS)score and the secondary outcomes included blood pressure,respiratory rate and pulse rate.All variables were evaluated at baseline(before bloodletting),0(after bloodletting immediately),15,30,50 and 80 min post bloodletting.Results At 80 min post bloodletting,the proportion of patients with improved consciousness in the bloodletting group was greater than the control group(P<0.05).In the separate analysis of moderate consciousness disturbance subgroup,bloodletting therapy benefited ischemic patients,and improved the eye and language response of GCS score at 15,30,50,80 min post bloodletting(P<0.05 or P<0.01).No significant differences were observed regarding the secondary outcomes between two groups(P>0.05).Conclusion The bloodletting puncture at HTWPs was safe and could improve conscious levels of ischemic stroke patients,highlighting a first-aid intervention for acute stroke.

Bloodletting Puncture at Hand Twelve Jing-Well Points Relieves Brain Edema after Severe Traumatic Brain Injury in Rats via Inhibiting MAPK Signaling Pathway

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 shamoperated(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.