Mechanisms of electroacupuncture relieves uterine smooth muscle spasm in dysmenorrhea rats based on Rho/ROCK signaling pathway. / 基于Rho/ROCKä¿¡å·é€šè·¯æŽ¢è®¨ç”µé’ˆä»»è„‰ã€ç£è„‰ç»ç©´ç¼“è§£ç±»ç—›ç»æ¨¡åž‹å¤§é¼ å­å®«å¹³æ»‘è‚Œç—‰æŒ›çš„æœºåˆ¶.

OBJECTIVES: To investigate the effect of electroacupuncture (EA) on Rho/Rho-associated coiled-coil-forming kinases (ROCK) signaling pathway of uterus tissue in rats with dysmenorrhea, so as to explore the underlying mechanism of EA treating primary dysmenorrhea (PD) and uterine smooth muscle spasm, and to observe whether there is a difference in the effect of meridian acupoints in Conception Vessel (CV) and Governer Vessel (GV). METHODS: Sixty female SD rats were randomly divided into saline, model, CV, GV, and non-acupoint groups, with 12 rats in each group. The dysmenorrhea model was established by subcutaneous injection of estradiol diphenhydrate combined with intraperitoneal injection of oxytocin (OT). EA (2 Hz) was applied to "Qihai" (CV6) and "Zhongji" (CV3) for CV group, "Mingmen" (GV4) and "Yaoshu" (GV2) for GV group, "non-acupoint 1" and "non-acupoint 3" on the left side for non-acupoint group, and manual acupuncture was applied to "Guanyuan" (CV4) for CV group, "Yaoyangguan" (GV3) for GV group, "non-acupoint 2" on the left side for non-acupoint group. The treatment was conducted for 20 min each time, once daily for 10 days. The writhing score was evaluated. The smooth myoelectric signals of rats' uterus in vivo were recorded by multi-channel physiological recorder. The uterine histopathological changes were observed by HE staining. The contents of prostaglandin F2α (PGF2α), OT and calcium ion (Ca2+) in uterine tissue of rats were detected by ELISA. The protein and mRNA expression levels of smooth muscle 22-α (SM22-α), RhoA and ROCKⅡ in uterine tissue were detected by Western blot and fluorescence quantitative PCR, respectively. RESULTS: Compared with the saline group, the writhing score of rats in the model group was increased (P<0.01), the amplitude voltage of uterine smooth muscle in vivo was elevated (P<0.01), the contents of PGF2α, OT and Ca2+, the protein and mRNA expression of SM22-α, RhoA and ROCK Ⅱ in uterine tissue were all increased (P<0.01). Compared with the model and the non-acupoint groups, the writhing scores of the CV and the GV groups were decreased (P<0.01, P<0.05), the amplitude voltage of uterine smooth muscle was decreased (P<0.01), the contents of PGF2α, OT and Ca2+ in uterine tissue were decreased (P<0.01, P<0.05), and the protein expression and mRNA expression of SM22-α, RhoA and ROCKⅡ in uterine tissue were decreased (P<0.01, P<0.05). HE staining showed extensive exfoliation of uterine intima with severe edema and increased glandular secretion in the model group, which was alleviated in the CV and GV groups. CONCLUSIONS: EA at acupoints of CV and GV can significantly reduce the writhing score, uterine smooth muscle amplitude voltage, pathological injury degree of uterus, and relieve spasm of uterine smooth muscle in dysmenorrhea rats, which may be related to its effect in regulating PGF2α and OT contents, inhibiting the Rho/ROCK signaling pathway, and reducing the SM22-α, RhoA, ROCKⅡ protein and mRNA expression, and Ca2+ content in uterine tissue.

Glycosides of Buyang Huanwu decoction inhibits inflammation associated with cerebral ischemia-reperfusion via the PINK1/Parkin mitophagy pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: A classic stroke formula is Buyang Huanwu Decoction (BYHWD), Glycosides are the pharmacological components found in BYHWD, which are utilized for the prevention and management of cerebral ischemia-reperfusion (CIR), as demonstrated in a previous study. Its neuroprotective properties are closely related to its ability to modulate inflammation, but its mechanism is as yet unclear. AIM OF THE STUDY: A research was undertaken to investigate the impact of glycosides on the inflammation of CIR through the PTEN-induced putative kinase-1 (PINK1)/Parkin mitophagy pathway. MATERIALS AND METHODS: Analyzing glycosides containing serum components was performed with ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). Glycosides were applied to rat of Middle cerebral artery occlusion/reperfusion (MCAO/R) model and primary neural cell of Oxygen glucose deprivation/reperfusion (OGD/R) model. The neuroprotective effect and the regulation of mitophagy of glycosides were evaluated through neural damage and PINK1/Parkin mitophagy activation. Moreover, the assessment of the relationship between glycosides regulation of mitophagy and its anti-inflammatory effects subsequent to mitophagy blockade was conducted by examining neural damage, PINK1/Parkin mitophagy activation, and levels of pyroptosis. RESULTS: (1) It was observed that the administration of glycosides resulted in a decrease in neurological function scores, a reduction in cerebral infarction volume, an increase in mitochondrial autophagosome, and the maintenance of a high expression status of light chain 3 (LC3) II/LC3Ⅰ protein. Additionally, there was a significant inhibition of p62 protein expression and an enhancement of PINK1 and Parkin protein expression. Furthermore, it was found that the effect of glycosides at a dosage of 0.128 g · kg-1 was significantly superior to that of glycosides at a dosage of 0.064 g · kg-1. Notably, the neuroprotective effect and inhibition of pyroptosis protein of glycosides at a dosage of 0.128 g · kg-1 were attenuated when mitochondrial autophagy was blocked. (2) Glycosides repaired cellular morphological damage, enhanced cell survival, and reduced Lactate dehydrogenase (LDH) leakage, with glycosides (2.36 µg·mL-1 and 4.72 µg·mL-1) neuronal protection being the strongest. Glycosides (4.72 µg·mL-1) maintained LC3II/LC3Ⅰ protein high expression state, inhibited p62 protein expression, and promoted PINK1 and Parkin protein expression, which was stronger than glycosides (2.36 µg·mL-1). The blockade of mitophagy resulted in a reduction of neuroprotection and inhibition of pyroptosis protein exerted by glycosides. CONCLUSION: Glycosides demonstrate the ability to hinder inflammation through the activation of the PINK1/Parkin mitophagy pathway, thereby leading to subsequent neuroprotective effects on CIR.

[The skin surface microcirculation of conception vessel, governor vessel and thoroughfare vessel in patients with primary dysmenorrhea].

OBJECTIVE: To observe the skin surface microcirculation of acupoints of conception vessel, governor vessel and thoroughfare vessel in patients with primary dysmenorrhea using laser speckle contrast imaging (LSCI), and provide acupoint selection basis of acupuncture-moxibustion for primary dysmenorrhea. METHODS: Ninety-nine healthy female college students with regular menstrual cycles (normal group) and 94 female college students with primary dysmenorrhea (dysmenorrhea group) were recruited. Before menstrual period, on the first day of menstruation, and on the third day after menstruation, LSCI was used to observe the surface microcirculation at the abdominal acupoints of conception vessel, i. e. Yinjiao (CV 7), Qihai (CV 6), Shimen (CV 5), Guanyuan (CV 4), Zhongji (CV 3) and Qugou (CV 2), acupoints of thoroughfare vessel, i. e. Huangshu (KI 16), Zhongzhu (KI 15), Siman (KI 14), Qixue (KI 13), Dahe (KI 12), Henggu (KI 11) and acupoints of lumbosacral region of governor vessel, i. e. Xuanshu (GV 5), Mingmen (GV 4), Yaoyangguan (GV 3), Yaoshu (GV 2) as well as two non-acupoints. RESULTS: Before menstrual period, there was no significant difference in the surface blood perfusion of the acupoints between the dysmenorrhea group and the normal group (P>0.05). On the first day of menstruation, the surface blood perfusion of Xuanshu (GV 5), Mingmen (GV 4), Yaoyangguan (GV 3) and right Huangshu (KI 16) in the dysmenorrhea group was higher than that in the normal group (P<0.05, P<0.01). On the third day after menstruation, the surface blood perfusion of the right Henggu (KI 11) in the dysmenorrhea group was lower than that in the normal group (P<0.05). CONCLUSION: In patients with primary dysmenorrhea, on the first day of menstruation, the surface blood perfusion of Xuanshu (GV 5), Mingmen (GV 4), Yaoyangguan (GV 3) of governor vessel, and the right Huangshu (KI 16) of thoroughfare vessel is increased, while on the third day after menstruation, the surface blood perfusion of the right Henggu (KI 11) of thoroughfare vessel is decreased. These findings might provide a basis for acupoint selection in the acupuncture-moxibustion treatment of primary dysmenorrhea.

Glycosides of Buyang Huanwu decoction inhibits pyroptosis associated with cerebral ischemia-reperfusion through Nrf2-mediated antioxidant signaling pathway both in vivo and in vitro.

BACKGROUND: Glycosides are the pharmacodynamic substances of Buyang Huanwu Decoction (BYHWD) and they exert a protective effect in the brain by inhibiting neuronal pyroptosis of cerebral ischemia-reperfusion (CIR). However, the mechanism by which glycosides regulate neuronal pyroptosis of CIR is still unclear. PURPOSE: A significant part of this study aimed to demonstrate whether glycosides have an anti-pyroptotic effect on CIR by nuclear factor erythroid 2-related factor (Nrf2)-mediated antioxidative mechanism. METHODS: Rats were used in vivo models of middle cerebral artery occlusion and reperfusion (MCAO/R). Neuroprotective effect of glycosides after Nrf2 inhibiting was observed by nerve function score, Nissl staining, Nrf2 fluorescence staining and pyroptotic proteins detection. SH-SY5Y cells were used in vitro models of oxygen-glucose deprivation/reperfusion (OGD/R). Glycosides was evaluated for their effects by measuring cell morphology, survival rate, lactate dehydrogenase (LDH) rate and expression of pyroptotic proteins. Nrf2 si-RNA 54-1 interference with lentivirus was used to create silenced Nrf2 SH-SY5Y cells (si-Nrf2-SH-SY5Y). Glycosides were evaluated on si-Con-SH-SY5Y and si-Nrf2-SH-SY5Y cells based on the expression of Nrf2 signaling pathway, pyroptotic proteins and cell damage manifestation. RESULTS: In vivo, glycosides significantly promoted the fluorescence level of nuclear Nrf2, added more Nissl bodies, reduced neurological function scores and inhibited the pyroptotic proteins level. In vitro, glycosides significantly repaired the morphological damage of cells, promoted the survival rate, reduced the LDH rate, inhibited the pyroptosis. Moreover, antioxidant activity of glycosides was enhanced via Nrf2 activation. Both Nrf2 blocking in vivo and Nrf2 silencing in vitro significantly weakened the pyroptosis inhibitory and neuroprotective effects of glycosides. CONCLUSION: These results suggested for the first time that glycosides inhibited neuronal pyroptosis by regulating the Nrf2-mediated antioxidant stress pathway, thereby exerting brain protection of CIR. As a result of this study, This study improved understanding of the pharmacodynamics and mechanism of BYHWD, as well as providing a Traditional Chinese Medicine (TCM) treatment strategy for CIR .

Acupuncture for the Treatment of Diarrhea-Predominant Irritable Bowel Syndrome: A Pilot Randomized Clinical Trial.

Importance: Acupuncture is a promising therapy for irritable bowel syndrome (IBS), but the use of subjective scales as an assessment is accompanied by high placebo response rates. Objectives: To preliminarily test the feasibility of using US Food and Drug Administration (FDA)-recommended end points to evaluate the efficacy of acupuncture in the treatment of IBS. Design, Setting, and Participants: This pilot, multicenter randomized clinical trial was conducted in 4 tertiary hospitals in China from July 1, 2020, to March 31, 2021, and 14-week data collection was completed in March 2021. Individuals with a diagnosis of IBS with diarrhea (IBS-D) were randomized to 1 of 3 groups, including 2 acupuncture groups (specific acupoints [SA] and nonspecific acupoints [NSA]) and a sham acupuncture group (non-acupoints [NA]) with a 1:1:1 ratio. Interventions: Patients in all groups received twelve 30-minute sessions over 4 consecutive weeks at 3 sessions per week (ideally every other day). Main Outcomes and Measures: The primary outcome was the response rate at week 4, which was defined as the proportion of patients whose worst abdominal pain score (score range, 0-10, with 0 indicating no pain and 10 indicating unbearable severe pain) decreased by at least 30% and the number of type 6 or 7 stool days decreased by 50% or greater. Results: Ninety patients (54 male [60.0%]; mean [SD] age, 34.5 [11.3] years) were enrolled, with 30 patients in each group. There were substantial improvements in the primary outcomes for all groups (composite response rates of 46.7% [95% CI, 28.8%-65.4%] in the SA group, 46.7% [95% CI, 28.8%-65.4%] in the NSA group, and 26.7% [95% CI, 13.0%-46.2%] in the NA group), although the difference between them was not statistically significant (P = .18). The response rates of adequate relief at week 4 were 64.3% (95% CI, 44.1%-80.7%) in the SA group, 62.1% (95% CI, 42.4%-78.7%) in the NSA group, and 55.2% (95% CI, 36.0%-73.0%) in the NA group (P = .76). Adverse events were reported in 2 patients (6.7%) in the SA group and 3 patients (10%) in NSA or NA group. Conclusions and Relevance: In this pilot randomized clinical trial, acupuncture in both the SA and NSA groups showed clinically meaningful improvement in IBS-D symptoms, although there were no significant differences among the 3 groups. These findings suggest that acupuncture is feasible and safe; a larger, sufficiently powered trial is needed to accurately assess efficacy. Trial Registration: Chinese Clinical Trial Registry: ChiCTR2000030670.

Effect of Auricular Acupoint Bloodletting plus Auricular Acupressure on Sleep Quality and Neuroendocrine Level in College Students with Primary Insomnia: A Randomized Controlled Trial.

OBJECTIVE: To evaluate the effects of auricular acupoint bloodletting (AB) and auricular acupressure (AA) on sleep quality and the levels of melatonin (MT), glutamic acid (Glu), and γ -aminobutyric acid (GABA) in college students with primary insomnia, and to explore the possible mechanism. METHODS: Totally 74 college students at Hebei University of Chinese Medicine with primary insomnia were selected from October 2019 to October 2020. All patients were assigned to AB+AA group (37 cases, received combination of AB and AA) and AA group (37 cases, received only AA on the same acupoints) by a random number table. Each group was treated twice a week for 4 weeks. The Pittsburgh Sleep Quality Index (PSQI) score, Chinese medicine (CM) syndrome score, total effective rate, serum concentrations of MT, Glu, and GABA, and Glu/GABA ratio were compared between the two groups after treatment and at follow-up. The safety of therapy was also evaluated. RESULTS: After 4-week treatment, the total scores of PSQI, each PSQI component score, and the CM syndrome scores in both groups all decreased (P<0.05); the serum MT concentrations in both groups all increased (P<0.05). The concentrations of Glu and GABA in the AB+AA group were significantly higher than those in the AA group after treatment (P<0.05). However, there was no significant difference in the ratio of Glu/GABA in both groups before and after treatment (P>0.05). At follow-up, the CM syndrome score in the AB+AA group was significantly lower than that in the AA group (P<0.05). There was no significant difference between the two groups in total effective rates and adverse events (P>0.05). CONCLUSIONS: Both AB+AA and AA can relieve insomnia symptoms, but a stronger long-term effect were observed for AB+AA. AB+AA can promote the secretion of MT, increase the levels of Glu and GABA more effective than AA, and regulate their imbalance, and thus it may be benificial for treating insomnia.

Effect of acupuncture for diarrhea-predominant irritable bowel syndrome: study protocol for a randomized clinical trial.

BACKGROUND: Diarrhea-predominant irritable bowel syndrome (IBS-D) is the most common subtype of IBS. Acupuncture is commonly used to treat IBS-D, but its effect is uncertain because of the poor quality of prior studies. This trial aims to evaluate the efficacy and safety of acupuncture treatment for IBS-D through comparisons with sham acupuncture. METHODS/DESIGN: This is a large-scale, multi-center, randomized, two-arm interventional clinical trial. Participants will take part in a total of 20 weeks of study, which contained 3 phases: 2-week screening, 6-week treatment, and 12-week follow-up. Based on the composite response rate of the primary endpoint in our pilot study (a sham acupuncture response rate of 27% and a true acupuncture of approximately 45%), 280 randomly allocated participants were planned. Eligible participants will be randomly assigned to the true acupuncture group and sham acupuncture group according to a ratio of 1:1, and a total of 15 sessions of treatment overall 6-week treatment period will be brought. The primary endpoint is a composite response rate at week 6, and the responder is defined as who responses in both abdominal pain intensity and stool consistency. Furthermore, composite response rates at other weeks, IBS Symptom Severity Scale, IBS Quality of Life, Adequate Relief scale, and individual IBS symptoms (abdominal pain, bloating, stool frequency) are chosen as secondary endpoints. DISCUSSION: This trial may provide high-quality evidence for the efficacy and safety of acupuncture in the treatment of IBS-D. The results of this study will be published in peer-reviewed journals. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR2100044762. Registered on 26 March 2021.

An evidence-based evaluation of Buyang Huanwu decoction for the treatment of the sequelae of stroke: A PRISMA-compliant systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Buyang Huanwu decoction (BYHWD) is a famous traditional Chinese formula that has been widely prescribed for sequelae of stroke in China. However, the efficacy and safety of BYHWD in treating sequelae of stroke have never been systematically evaluated. PURPOSE: To evaluate the effectiveness and safety of BYHWD in the treatment of sequelae of stroke. STUDY DESIGN: A Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA)-compliant systematic review and meta-analysis of randomized clinical trials (RCTs). MATERIALS AND METHODS: A systematic review and meta-analysis was performed in accordance with the PRISMA guidelines. Five common electronic databases were searched for relevant RCTs from their inception until May 20, 2022. The Cochrane risk-of-bias tool was used to evaluate the methodological quality and the risk of bias of the included RCTs. Review Manager 5.4 was used to analyse all the data obtained. The clinical effective rate (CER) was the primary outcome, and National Institutes of Health Stroke Scale (NIHSS) and Fugl-Meyer Assessment (FMA) scores were the secondary outcomes. The Grading of Recommendations, Assessment, Development and Evaluations (GRADE) system was used to evaluate the quality of evidence for each outcome. RESULTS: Thirty-two clinical studies that recruited 2,527 eligible patients were included in this meta-analysis. The results of the meta-analysis suggested that compared with conventional treatment alone, the addition of BYHWD significantly improved the CER (RR = 1.24, 95% CI: 1.20-1.29, p < 0.00001), decreased the NIHSS score (MD = -5.42, 95% CI: -5.87-4.97, p < 0.00001), and increased the FMA score (MD = 17.28, 95% CI: 15.12-19.45, p < 0.00001). There were no reported adverse events in the included studies. Most results were robust, and the quality of evidence was moderate. CONCLUSION: Our study is the first meta-analysis of RCTs evaluating the effects of BYHWD on sequelae of stroke. The addition of BYHWD to conventional treatment for sequelae of stroke significantly improved the CER and promoted neurological rehabilitation in patients, and there were no reported adverse events associated with this combination therapy. The findings of our study support the use of BYHWD as an adjunct treatment to conventional treatment in this clinical population. However, due to the limitations of the included clinical trials, high-quality clinical trials with longer follow-ups are needed to assess the long-term effectiveness and safety of BYHWD for treating the sequelae of stroke.

[Clinical observation on different frequency of auricular bloodletting combined with auricular point sticking for acne vulgaris].

OBJECTIVE: To observe the clinical efficacy of different treatment frequency of auricular bloodletting combined with auricular point sticking for acne vulgaris. METHODS: A total of 90 patients with acne vulgaris were randomized into a treatment group 1 (30 cases, 2 cases dropped off), a treatment group 2 (30 cases, 4 cases dropped off) and a treatment group 3 (30 cases, 5 cases dropped off). Combination therapy of auricular bloodletting and auricular point sticking at Fei (CO14), Shenmen (TF4), Neifenmi (CO18) and Shenshangxian (TG2p) were given once a week, twice a week and 3 times a week in the treatment group 1, the treatment group 2 and the treatment group 3 respectively, 4 weeks were as one course and totally 3 courses were required in the 3 groups. Before treatment and after 1, 2, 3 courses of treatment, the scores of global acne grading system (GAGS), skin lesion and quality of life-acne (QoL-Acne) were observed, the clinical efficacy was evaluated after 3 courses of treatment and the recurrence rate was evaluated in follow-up of 1 month after treatment in the 3 groups. RESULTS: Compared before treatment, the scores of GAGS and skin lesion were decreased at each time point in the 3 groups (P<0.05), the QoL-Acne scores were increased after 1 course of treatment in the treatment group 3 and after 2, 3 courses of treatment in the 3 groups (P<0.05). There were no statistical differences in scores of GAGS, skin lesion and QoL-Acne among the 3 groups (P>0.05). There were no statistical differences in effective rate and recurrence rate among the 3 groups (P>0.05). CONCLUSION: Different frequency of auricular bloodletting combined with auricular point sticking have similar efficacy in treating acne vulgaris, all can improve the skin lesion and quality of life in patients with acne vulgaris, and have a cumulative effect and good long-term curative effect.

[Comparative study on the academic thought of bloodletting for expelling pathogens between Jin-Yuan Dynasties and Ming-Qing Dynasties].

The relevant provisions of bloodletting for expelling pathogens are collected from the works of the medical representative scholars in Jin-Yuan Dynasties and Ming-Qing Dynasties respectively to construct the databases of bloodletting for expelling pathogens of Jin-Yuan Dynasties and Ming-Qing Dynasties. Using frequency analysis, the bloodletting device, bloodletting location, bloodletting volume, the related pathogens and indications are compared between these two times so that the evidences could be provided for the inheritance and development of the academic thought of bloodletting for expelling pathogens. It is found that the three-edge needle is the most commonly used device for bloodletting in Jin-Yuan Dynasties and Ming-Qing Dynasties and yang meridians and local affected area are generally selected for bloodletting. The range of meridian and acupoint selection in Ming-Qing Dynasties are more extensive than those in Jin-Yuan Dynasties, while bloodletting volume is less than that in Jin-Yuan Dynasties. In Jin-Yuan Dynasties, bloodletting therapy is mainly for expelling exogenous pathogens i.e. heat, fire and wind, while, in Ming-Qing Dynasties, this therapy is specially for clearing heat and removing stasis. The disorders of internal medicine are often treated with such therapy in these two dynasties. But, compared with Jin-Yuan Dynasties, the indication of bloodletting therapy is expanded gradually in Ming-Qing Dynasties. It is shown that the academic thought of bloodletting for expelling pathogens in Jin-Yuan Dynasties is inherited and developed from Ming-Qing Dynasties.

Acupuncture for the treatment of diarrheal-predominant irritable bowel syndrome: study protocol for a pilot randomized controlled trial.

BACKGROUND: Irritable bowel syndrome (IBS) is one of the most common functional gastrointestinal diseases. Although acupuncture has become a common alternative therapy for IBS, there is insufficient evidence for its effectiveness. This study was designed to assess the efficacy and feasibility of acupuncture in the treatment of IBS. METHODS/DESIGN: This is a multicenter randomized controlled clinical trial. According to the ratio of 1:1:1, 90 patients with irritable bowel syndrome will be randomly divided into specific acupoints (SA) group, non-specific acupoints (NSA) group, and non-acupoints (NA) group. All patients will be treated with acupuncture 12 times within 4 weeks and followed up for 8 weeks. The primary outcome is the response rate, the percentage of patients whose average value of worst abdominal pain is 30% better and the days of loose stool is 50% less than the baseline, at week 4 after randomization. The secondary outcomes include the response rates at other time points, IBS Symptom Severity Scale (IBS-SSS), Patient Health Questionnaire-9 depression scale (PHQ-9), IBS-Quality of Life scale (IBS-QOL), IBS Adequate Relief (IBS-AR), Abdominal Pain Score, Abdominal Bloating Score, Bristol Stool Score (BBS), blinding assessment, and credibility evaluation. Adverse events will be monitored and recorded during the trial. TRIAL REGISTRATION: Chictr.org.cn ChiCTR2000030670. Registered on 9 March 2020.

[Changes of microcirculation in superficial tissues of acupoint regions of lower leg during menstrual cycle in healthy young girls].

OBJECTIVE: To observe changes of microcirculation in the superficial regions of acupoints of the three Yin meridians of foot during the menstrual cycle in young college students, so as to provide experimental evidence for explaining the saying of traditional Chinese medicine that acupoints reflect the state of physiological and pathological activities of the internal organs. METHODS: Ninety healthy female volunteer college students were recruited in the present study. The subjects were asked to take a supine position on an examination couch to expose the Yuan-primary acupoints Taixi (KI3), Taibai (SP3) and Taichong (LR3), and Xi-cleft acupoints Shuiquan (KI5), Diji (SP8) and Zhongdu (LR6) which are related to the uterus of the three Yin meridians of foot， the crossing acupoints of the three Yin meridians of foot Sanyinjiao (SP6), non-specific acupoint of the Spleen meridian Xuehai (SP10), non-related meridian acupoint Xuanzhong (GB39) and non-meridian-non-acupoint (being at the same level of GB39, between the Stomach and Gallbladder meridians on the lateral aspect of the lower leg). The laser speckle blood flow imaging technique was used to detect the state of microcirculation (average blood perfusion volume) during menstrual, follicular, ovulatory and luteal phases. RESULTS: The average blood perfusion volume (ABPV) of the right SP8 region was significantly lower in the menstrual phase than in the ovulation and luteal phases (P<0.05), and in the follicular phase than in the ovulation phase (P<0.05). In the left LR6 region, the ABPV was obviously lower in the menstrual period than in the follicular, ovulation and luteal phases (P<0.05). In the SP8, SP6, KI3 and KI5 regions, the ABPV was significantly higher in each of the 4 phases on the left side than on the right side (P<0.05). In the right GB39, the ABPV in each of the 4 phases was apparently higher on the right side than on the left side (P<0.05). In the SP10 region, the ABPV was considerably higher on the left side than on the right side in the ovulation period (P<0.05). The ABPV of SP3 in the menstrual phase and that of the non-acupoint in the luteal phase were significantly higher on the right side than on the left side (P<0.05). CONCLUSION: The blood perfusion of microcirculation in the superficial tissues of Xi-cleft acupoints SP8 and LR6 is obviously lower in the menstrual phase than in both of the ovulation and luteal phases in healthy young college students, which may specifically reflect the periodical activities of the uterus in the physiological state, and provide a basis of acupoint selection for menstrual healthy care.

[Progress of researches on electrical characteristics of meridians and acupoints in recent 10 years].

The electrical characteristics of meridians and acupoints have been being one of the hot topics discussed by scholars, because of many influencing factors and inconsistent research results. In the present article, we collected papers published in Chinese or English in recent 10 years from PubMed, CNKI, and VIP databases by using key words of"acupuncture""meridian""resistance""capacitance""electrodermal"and"impedance". Then, we make an analysis about its development from three aspects, 1) manifestations of electrical properties of acupoints (low dermal resistance under pathological state, imbalance between left and right sides of the synonym acupoints), 2) clinical application of acupoint electrical characteristics for diagnosis of clinical disorders and evaluation of therapeutic effect, and 3) current situations of development of meridian detection instruments based on electrical characteristics (improving stability of instruments，expansion of the detected regions and measuring time, realization of visualization, and operational automation). In the process of coming studies, we highly recommend that more attention should be paid to the acupoints' "dynamic changes"， and forming a standardized research plan, strengthening domestic and international academic exchanges and cooperation are definitely necessary in order to validate the objectivity and specificity of electrical characteristics of meridians and acupoints, further expanding its clinical application.

[Regularity of body surface resistance at the acupoints of three yin meridians of the foot in reflecting physiological changes of the uterus].

OBJECTIVE: To investigate the specific regularity of body surface resistance at different acupoints of the meridians associated with the uterus in reflecting menstrual cycle by observing the change in body surface resistance at source points, cleft points, confluent points, and non-specific points of three yin meridians of the foot in different menstrual cycles in normal female college students, to lay a foundation for the in-depth research on the mechanism of acupoints reflecting the function of zang-fu, and to provide a reference for the clinical and scientific research on the biophysical characteristics of menstrual cycle-related acupoints in normal female. METHODS: A total of 90 normal female college students were recruited. The source points, cleftpoints, confluent points, and non-specific points of three yin meridians of the foot which were located in the adjacent spinal segments of the uterus were selected, and body surface resistance was monitored for 30 consecutive minutes at the same time-points of menstrual phase, follicular phase, ovulation phase, and luteal phase to observe the change in the resistance of each acupoint during the menstrual cycle. RESULTS: There was no significant change in the resistance value of unilateral acupoints during the menstrual cycle (P>0.05). In the same period, there was no significant difference in resistance value between unilateral three source points, three cleftpoints, different acupoints of the spleen meridian, and different acupoints of lumbar 4 dermatomere (P>0.05). As for the comparison of resistance of the same acupoint at the left and right sides, Taichong (LR3) at the left side had a higher resistance value than that at the right side in the menstrual phase (P<0.05); Taibai (SP3) at the left side had a higher resistance value than that at the right side in the ovulation phase (P<0.05); Zhongdu (LR6) at the left side had a higher resistance value than that at the right side in the follicular phase, the ovulation phase, and the luteal phase (P<0.05); Taixi (KI3), Diji (SP8), and Sanyinjiao (SP6) at the left side had a higher resistance value than those at the right side in the menstrual phase, the follicular phase, the ovulation phase, and the luteal phase (P<0.05); Shuiquan (KI5) at the left side had a lower resistance value than that at the right side in these four phases (P<0.05). CONCLUSION: The change trend of the resistance of the acupoints at the left and right sides associated with the three yin meridians of the foot can specifically reflect the change in qi and blood in the uterus during the menstrual cycle. The change trend of the source and cleft points of the liver meridian in the menstrual phase is different from that in the other phases, and the change trend of SP3, a source point of the spleen meridian, in the ovulation phase is different from that in the other phases, which suggests the specificity of meridian points in reflecting function. The mechanism by which meridian points reflect the function of zang-fu is associated with the meridian points and the spinal cord segments of zang-fu, and meanwhile, it has a specific relationship with the meridians to which meridian points belong and the attributes of acupoints.

Neuroprotective effect of glycosides in Buyang Huanwu Decoction on pyroptosis following cerebral ischemia-reperfusion injury in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Buyang Huanwu Decoction (BYHWD) is used in classical traditional Chinese medicine to prevent and treat cerebral ischemia. Glycosides, which are effective components extracted from BYHWD, mainly include astragaloside IV, paeoniflorin, and amygdalin. These glycosides are the primary pharmacologically effective constituents of BYHWD that act against cerebral ischemic nerve injury; however, the mechanism of action of BYHWD is still unclear. AIM OF THE STUDY: The present study aimed to determine the effect of BYHWD glycosides on pyroptosis after cerebral ischemia reperfusion injury and explore whether its mechanism involves the classical pyroptosis pathway mediated by NLRP3. MATERIAL AND METHODS: Adult male Sprague-Dawley rats (n = 140) were randomly divided into seven groups: sham, cerebral ischemia and reperfusion (I/R), glycosides (0.064 g/kg, 0.128 g/kg, and 0.256 g/kg), BYHWD, and AC-YVAD-CMK (caspase-1 inhibitor). A rat model of cerebral I/R was established via classic middle cerebral artery occlusion (MCAO) for 2 h, followed by 24-h reperfusion. Neurological function was estimated using neurological defect scores. Brain infarct volumes were determined by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and nerve cell damage was evaluated by Nissl staining. Pyroptosis was detected using TUNEL and caspase-1 immunofluorescence double staining. Protein expression of NLRP3, ASC, caspase-1, pro-caspase-1, and IL-1ß was analyzed using Western blot analysis. RESULTS: Glycosides improved neurological dysfunction, alleviated neuronal damage, and inhibited neuronal pyroptosis. The 0.128 g/kg glycosides group showed the most significant effects. Furthermore, we observed that this group showed significant inhibition of the expression of NLRP3, ASC, pro-caspase-1, caspase-1, and IL-1ß proteins of the NLRP3-mediated classical pathway of pyroptosis. CONCLUSIONS: Glycosides exert neuroprotective effects by inhibiting pyroptosis of neurons after cerebral I/R injury. The underlying mechanism of action is closely related to the regulation of the classical pyroptosis pathway by NLRP3.

Influence of de qi on the immediate analgesic effect of SP6 acupuncture in patients with primary dysmenorrhoea and cold and dampness stagnation: a multicentre randomised controlled trial.

OBJECTIVE: The aim of this multicentre randomised controlled trial was to investigate the contribution of de qi to the immediate analgesic effect of acupuncture in patients with primary dysmenorrhoea and the specific traditional Chinese medicine diagnosis cold and dampness stagnation. METHOD: Eighty-eight patients with primary dysmenorrhoea and cold and dampness stagnation were randomly assigned to de qi (n=43) or no de qi (n=45) groups and underwent 30 min of SP6 acupuncture. The de qi group received deep needling at SP6 with manipulation using thick needles; the no de qi group received shallow needling with no manipulation using thin needles. In both groups the pain scores and actual de qi sensation were evaluated using a visual analogue scale for pain (VAS-P) and the acupuncture de qi clinical assessment scale (ADCAS), respectively. RESULTS: Both groups showed reductions in VAS-P, with no signficant differences between groups. ADCAS scores showed 43/43 and 25/45 patients in de qi and no de qi groups, respectively, actually experienced de qi sensation. Independent of original group allocation, VAS-P reductions associated with actual de qi (n=68) were greater than those without (28.4±18.19 mm vs 14.6±12.28 mm, p=0.008). CONCLUSIONS: This study showed no significant difference in VAS-P scores in patients with primary dysmenorrhoea and cold and dampness stagnation immediately after SP6 acupuncture designed to induce or avoid de qi sensation. Both treatments significantly reduced VAS-P relative to baseline. Irrespective of group allocation, patients experiencing actual de qi sensation demonstrated larger reductions in pain score relative to those without, suggesting greater analgesic effects. TRIAL REGISTRATION NUMBER: Chinese Clinical Trial Registry (ChiCTR-TRC-13003086); Results.

Buyang Huanwu Decoction Ameliorates Poststroke Depression via Promoting Neurotrophic Pathway Mediated Neuroprotection and Neurogenesis.

Objective. The aim of the present research is to investigate the therapeutic effect of Buyang Huanwu Decoction (BHD) in poststroke depression (PSD) animal model and illustrate its underlying mechanism via promoting neurotrophic pathway mediated neuroprotection and neurogenesis. Methods. To induce PSD rat model, isolation housed rats that received middle cerebral artery occlusion (MCAO) surgery successively suffered from chronic mild stress (CMS) treatment for consecutive twenty-one days. Meanwhile, rats were correspondingly given vehicle, BHD, and fluoxetine. Then, neurologic function was scored and depressive-like behaviors were assessed by sucrose preference test, locomotor activity, novelty-suppressed feeding test, and forced swim test. Thereafter, the neuroprotection and neurogenesis related molecular markers and signaling were detected. Results. We firstly observed a significant neurological function recovery and antidepressants effect of BHD after MCAO together with CMS treatment. Our study also found that treatment with BHD and fluoxetine can significantly rescue neurons from apoptosis and promote neurogenesis in the CA3 and DG regions in the hippocampus. Notably, BHD and fluoxetine treatment can activate BDNF/ERK/CREB signaling. Conclusion. The results suggest that BHD is a promising candidate for treating PSD. Its curative effects can be attributed to neurotrophic pathway mediated neuroprotection and neurogenesis.

The role of oxidative stress in DNA damage in pancreatic ß cells induced by di-(2-ethylhexyl) phthalate.

Di(2-ethyhexyl) phthalate (DEHP) is commonly used as a plasticizer, which loosely binds to plastic materials and easily leaches out of these products and enters into the environment. Exposure to DEHP can impair pancreatic beta cells (INS-1 cells)function, which is associated with Insulin Resistance (IR) and type 2 diabetes. However, the mechanism of how DEHP leads to Insulin Resistance is unknown. Our results showed that the cell viability of INS-1 cells exposed to DEHP (0-1600 µM) were decreased in a concentration-dependent manner. DEHP caused significant increases of DNA migration and oxidative damage in INS-1 cells. Lysosomal membrane permeability was increased and mitochondrial membrane potential was reduced after INS-1 cells treated with DEHP. DEHP was also shown to induce ROS production and cause GSH depletion in INS-1 cells. DEHP brought a significant decrease in super oxide dismutase (SOD) and led to accumulation of malondialdehyde (MDA) in the INS-1 cells. DEHP increased significantly the expression of P53 and ATM gene of INS-1 cell at high dose levels. Simultaneously, Pyrroloquinoline Quinone (PQQ), an antioxidant, and alcohol were used in the study to determine their effects on DEHP-induced INS-1 cells damage. PQQ could protect the INS-1 cells from the damage induced by DEHP to some extent, while alcohol aggravated the toxic effects of DEHP. These results indicate that DEHP-mediated INS-1 cell dysfunction through a lysosomal-mitochondrial pathway, involving oxidative stress and p53 and ATM activation.

[Effect of Buyang Huanwu decoction and its simple prescription (Naojian tablet) on CDK4/Cyclin D1 expression of rats with cerebral ischemia].

To evaluate the regulating effect of Buyang Huanwu decoction and its simple prescription (Naojian tablet) on CDK4/Cyclin D1 expression in hippocampus tissues of rats with cerebral ischemia, SD rats were divided into the sham-operation group, the model group, the Buyang Huanwu decoction group (ig, 3.15 g · kg⁻¹) and the simple prescription group (ig, 2.41 g · kg⁻¹). Each group was further divided into five subgroups based on time points after the administration, i. e. 1 d, 3 d, 7 d, 14 d and 28 d, respectively. CDK4/Cyclin D1 expressions of the group at different time points were examined by using immunohistochemistry and real-time qPCR. According to the results, the cerebral ischemia model group showed higher CDK4/Cyclin D1 expression than the sham-operation groups (P < 0.05), suggesting that the cell cycle signal pathway would be activated by the cerebral ischemic injury. Both Buyang Huanwu decoction and simple prescription groups showed significantly lower cyclin expression than the model group at 3 d, 7 d, 14 d, 28 d (P < 0.05), indicating both Buyang Huanwu decoction and its simple prescription could play the neuroprotective effect through the cell cycle signal pathway.