Effect of electroacupuncture on myocardial electrical remodeling in rats with acute myocardial infarction. / ç”µé’ˆå¯¹æ€¥æ€§å¿ƒè‚Œæ¢—æ­»å¤§é¼ å¿ƒè‚Œç”µé‡æž„çš„å½±å“åŠå ¶æœºåˆ¶ç ”ç©¶.

OBJECTIVES: To investigate the mechanism of electroacupuncture (EA) at "Neiguan" (PC6) in impro-ving myocardial electrical remodeling in rats with acute myocardial infarction (AMI) by enhancing transient outward potassium current. METHODS: A total of 30 male SD rats were randomly divided into control, model and EA groups, with 10 rats in each group. The AMI model was established by subcutaneous injection with isoprenaline (ISO, 85 mg/kg). EA was applied to left PC6 for 20 min, once daily for 5 days. Electrocardiogram (ECG) was recorded after treatment. TTC staining was used to observe myocardial necrosis. HE staining was used to observe the pathological morphology of myocardial tissue and measure the cross-sectional area of myocardium. Potassium ion-related genes in myocardial tissue were detected by RNA sequencing. The mRNA and protein expressions of Kchip2 and Kv4.2 in myocardial tissue were detected by real-time fluorescence quantitative PCR and Western blot, respectively. RESULTS: Compared with the control group, cardiomyocyte cross-sectional area in the model group was significantly increased (P<0.01), the ST segment was significantly elevated (P<0.01), and QT, QTc, QTd and QTcd were all significantly increased (P<0.05, P<0.01). After EA treatment, cardiomyocyte cross-sectional area was significantly decreased (P<0.01), the ST segment was significantly reduced (P<0.01), and the QT, QTc, QTcd and QTd were significantly decreased (P<0.01, P<0.05). RNA sequencing results showed that a total of 20 potassium ion-related genes co-expressed by the 3 groups were identified. Among them, Kchip2 expression was up-regulated most notablely in the EA group. Compared with the control group, the mRNA and protein expressions of Kchip2 and Kv4.2 in the myocardial tissue of the model group were significantly decreased (P<0.01, P<0.05), while those were increased in the EA group (P<0.01, P<0.05). CONCLUSIONS: EA may improve myocardial electrical remodeling in rats with myocardial infarction, which may be related to its functions in up-regulating the expressions of Kchip2 and Kv4.2.

Folium Hibisci Mutabilis extract, a potent autophagy enhancer, exhibits neuroprotective properties in multiple models of neurodegenerative diseases.

BACKGROUND: Protein aggregates are considered key pathological features in neurodegenerative diseases (NDs). The induction of autophagy can effectively promote the clearance of ND-related misfolded proteins. OBJECTIVE: In this study, we aimed to screen natural autophagy enhancers from traditional Chinese medicines (TCMs) presenting potent neuroprotective potential in multiple ND models. METHODS: The autophagy enhancers were broadly screened in our established herbal extract library using the transgenic Caenorhabditis elegans (C. elegans) DA2123 strain. The neuroprotective effects of the identified autophagy enhancers were evaluated in multiple C. elegans ND models by measuring Aß-, Tau-, α-synuclein-, and polyQ40-induced pathologies. In addition, PC-12 cells and 3 × Tg-AD mice were employed to further validate the neuroprotective ability of the identified autophagy enhancers, both in vitro and in vivo. Furthermore, RNAi bacteria and autophagy inhibitors were used to evaluate whether the observed effects of the identified autophagy enhancers were mediated by the autophagy-activated pathway. RESULTS: The ethanol extract of Folium Hibisci Mutabilis (FHME) was found to significantly increase GFP::LGG-1-positive puncta in the DA2123 worms. FHME treatment markedly inhibited Aß, α-synuclein, and polyQ40, as well as prolonging the lifespan and improving the behaviors of C. elegans, while siRNA targeting four key autophagy genes partly abrogated the protective roles of FHME in C. elegans. Additionally, FHME decreased the expression of AD-related proteins and restored cell viability in PC-12 cells, which were canceled by cotreatment with 3-methyladenine (3-MA) or bafilomycin A1 (Baf). Moreover, FHME ameliorated AD-like cognitive impairment and pathology, as well as activating autophagy in 3 × Tg-AD mice. CONCLUSION: FHME was successfully screened from our natural product library as a potent autophagy enhancer that exhibits a neuroprotective effect in multiple ND models across species through the induction of autophagy. These findings offer a new and reliable strategy for screening autophagy inducers, as well as providing evidence that FHME may serve as a possible therapeutic agent for NDs.

Effects of electroacupuncture on rats with cognitive impairment: An iTRAQ-based proteomics analysis.

OBJECTIVE: The study explores the effects of electroacupuncture (EA) at the governing vessel (GV) on proteomic changes in the hippocampus of rats with cognitive impairment. METHODS: Healthy male rats were randomly divided into 3 groups: sham, model and EA. Cognitive impairment was induced by left middle cerebral artery occlusion in the model and EA groups. Rats in the EA group were treated with EA at Shenting (GV24) and Baihui (GV20) for 7 d. Neurological deficit was scored using the Longa scale, the learning and memory ability was detected using the Morris water maze (MWM) test, and the proteomic profiling in the hippocampus was analyzed using protein-labeling technology based on the isobaric tag for relative and absolute quantitation (iTRAQ). The Western blot (WB) analysis was used to detect the proteins and validate the results of iTRAQ. RESULTS: Compared with the model group, the neurological deficit score was significantly reduced, and the escape latency in the MWM test was significantly shortened, while the number of platform crossings increased in the EA group. A total of 2872 proteins were identified by iTRAQ. Differentially expressed proteins (DEPs) were identified between different groups: 92 proteins were upregulated and 103 were downregulated in the model group compared with the sham group, while 142 proteins were upregulated and 126 were downregulated in the EA group compared with the model group. Most of the DEPs were involved in oxidative phosphorylation, glycolipid metabolism and synaptic transmission. Furthermore, we also verified 4 DEPs using WB technology. Although the WB results were not exactly the same as the iTRAQ results, the expression trends of the DEPs were consistent. The upregulation of heat-shock protein ß1 (Hspb1) was the highest in the EA group compared to the model group. CONCLUSION: EA can effect proteomic changes in the hippocampus of rats with cognitive impairment. Hspb1 may be involved in the molecular mechanism by which acupuncture improves cognitive impairment.

Integrated Chemical Characterization, Network Pharmacology and Transcriptomics to Explore the Mechanism of Sesquiterpenoids Isolated from Gynura divaricata (L.) DC. against Chronic Myelogenous Leukemia.

Chronic myelogenous leukemia (CML) is a serious threat to human health, while drugs for CML are limited. Herbal medicines with structural diversity, low toxicity and low drug resistance are always the most important source for drug discoveries. Gynura divaricata (L.) DC. is a well-known herbal medicine whose non-alkaline ingredients (GD-NAIs) were isolated. The GD-NAIs demonstrated potential anti-CML activity in our preliminary screening tests. However, the chemical components and underlying mechanism are still unknown. In this study, GD-NAIs were tentatively characterized using UHPLC-HRMS combined with molecular networking, which were composed of 75 sesquiterpenoids. Then, the anti-CML activities of GD-NAIs were evaluated and demonstrated significant suppression of proliferation and promotion of apoptosis in K562 cells. Furthermore, the mechanism of GD-NAIs against CML were elucidated using network pharmacology combined with RNA sequencing. Four sesquiterpenoids would be the main active ingredients of GD-NAIs against CML, which could regulate PD-L1 expression and the PD-1 checkpoint pathway in cancer, PI3K/AKT, JAK/STAT, TGF-ß, estrogen, Notch and Wnt signaling pathways. In conclusion, our study reveals the composition of GD-NAIs, confirms its anti-CML activity and elucidates their underlying mechanism, which is a potential countermeasure for the treatment of CML.

A comprehensive review of Rubia cordifolia L.: Traditional uses, phytochemistry, pharmacological activities, and clinical applications.

Rubia cordifolia (family: Rubiaceae) L (R. cordifolia) is a perennial botanical drug climbing vine. As the main part of the traditional Chinese medicine, the rhizome has a long history. A great number of literary studies have reported that it can be used for the improvement of blood circulation, hemostasis, activation of collaterals, etc. When it comes to the wide application of R. cordifolia in traditional medicine, we systematically review its traditional uses, phytochemistry and pharmacological effects. Literatures were systematically searched using several scientific databases, including China National Knowledge Infrastructure (CNKI), Baidu Scholar, PubMed, Web of Science, and other professional websites. Kew Botanical Garden and the iPlant were used for obtaining the scientific names and plant images of R. cordifolia. In addition, other information was also gathered from books including traditional Chinese herbal medicine, the Chinese Pharmacopoeia, and Chinese Materia Medica. So far, many prescriptions containing R. cordifolia have been widely used in the clinical treatment of abnormal uterine bleeding, primary dysmenorrhea and other gynecological diseases, allergic purpura, renal hemorrhage and other diseases. The phytochemistry studies have reported that more than 100 compounds are found in R. cordifolia, such as bicyclic peptides, terpenes, polysaccharides, trace elements, flavonoids, and quinones. Among them, quinones and peptides are the types of components with the highest contents in R. cordifolia. The modern pharmacological studies have revealed that R. cordifolia and its derived components have anti-tumor, anti-oxidative, anti-platelet aggregation, and anti-inflammatory effects. However, most studies are preclinical. The pharmacological mechanism of R. cordifolia has not been thoroughly studied. In addition, there are few pharmacokinetic and toxicity studies of R. cordifolia, therefore the clinical safety data for R. cordifolia is lacking. To sum up, this review for the first time summarizes a systemic and integrated traditional uses, chemical compositions, pharmacological actions and clinical applications of R. cordifolia, which provides the novel and full-scale insight for the drug development, medicinal value, and application of R. cordifolia in the future.

Chemical characterization and DPP-IV inhibitory activity evaluation of tripeptides from Gynura divaricata (L.) DC.

ETHNOPHARMACOLOGICAL RELEVANCE: Gynura divaricata (L.) DC. (GD), a herbal medicine, has been used for the prevention and treatment of hyperglycemia in China. However, hypoglycemic ingredients within GD have not yet been well studied. AIM OF THE STUDY: The aim of this study was to explore undiscovered compounds with dipeptidyl peptidase IV (DPP-IV) inhibitory activity within GD. MATERIALS AND METHODS: A four-step strategy was developed to explore undiscovered DPP-IV inhibitors within GD. First, the components were preliminarily characterized using UHPLC-HRMS combined with a library search. Second, preliminarily characterized compounds were searched for potential bioactivity. Third, a mixture of these preliminarily characterized compounds was isolated and thoroughly characterized based on fragmentation patterns associated with molecular networking. Fourth, the activities of these compounds were verified using DPP-IV inhibitory assay and molecular docking. RESULTS: Diprotin A, a tripeptide inhibitor against DPP-IV, was identified. Thereafter, a mixture of twenty-five diprotin A analogs was isolated and characterized, which exhibited IC50 of 0.40 mg/mL for DPP-IV. Molecular docking results also confirmed the interactions between the tripeptide analogs and DPP-IV mainly via H-bonds and hydrophobic interactions. CONCLUSIONS: This is the first report of DPP-IV inhibitors within GD. These findings demonstrate that the extract of GD might be beneficial for the treatment of type 2 diabetes mellitus, and is expected to promote further development and utilization of GD in herbal medicine.

Treatment of pulmonary fibrosis in one convalescent patient with corona virus disease 2019 by oral traditional Chinese medicine decoction: A case report.

After one-month of oral treatment with traditional Chinese medicine decoction, without using other drugs, the lung inflammatory exudate, pulmonary fibrosis and quality of life of a 61-year-old female patient with corona virus disease 2019 (COVID-19) were significantly improved. No recurrence or deterioration of the patient's condition was found within seven weeks of treatment and follow-up, and no adverse events occurred, indicating that oral Chinese medicine decoction was able to improve the pulmonary inflammation and fibrosis in a patient recovering from COVID-19, but further research is still needed.

Effects of progressive muscle relaxation training on negative emotions and sleep quality in COVID-19 patients: A clinical observational study.

This study investigates the effect of progressive muscle relaxation training on negative mood and sleep quality in Coronavirus Pneumonia (COVID-19) patients.COVID-19 is an emerging infectious disease, and there is still uncertainty about when the outbreak will be contained and the effectiveness of treatments. Considering that this disease is highly contagious, patients need to be treated in isolation. This may lead to psychological symptoms such as anxiety and depression, and even sleep problems.This study is a clinical observation study.Participants included 79 COVID-19 patients admitted to a designated hospital for COVID-19 patients in Wuhan from February to March, 2020. Patients were selected and assigned to the control group and the observation group according to their wishes, with 40 and 39 cases in each group, respectively. The control group received routine treatment and nursing, and the observation group received progressive muscle relaxation training, in addition to the routine treatment and nursing. We compared scores of the Pittsburgh Sleep Quality Index Scale (PSQI), the Generalized Anxiety Disorder (GAD-7), and the Patient Health Questionnaire (PHQ-9) before and after the intervention.There was no significant difference in PSQI, GAD-7, and PHQ-9 scores between the control group and the observation group before the intervention (P > .05). After the intervention, the difference in scores of PSQI, GAD-7, and PHQ-9 in the 2 groups were statistically significant (P < .05).Progressive muscle relaxation training can significantly reduce anxiety and depression and improve sleep quality in COVID-19 patients during isolation treatment.Progressive muscle relaxation training was shown to improve the treatment effect of patients and is worthy of clinical promotion.

[Changes of Skin Temperature of Acupoint Regions of the Governor Meridian after Moxibustion Stimulation].

OBJECTIVE: To observe changes of skin temperature along the Governor Vessel (GV) after moxibustion stimulation. METHODS: A total of 36 healthy volunteer subjects were recruited in the present study. The skin temperature of Mingmen (GV 4), Dazhui (GV 14), Shenzhu (GV 12), Zhiyang (GV 9), Zhongshu (GV 7), Yaoyangguan (GV 3) regions and their bilateral control sites (1.5 cun lateral to GV 4) was detected by using a Thermo Tracer TH 9100. RESULTS: Following moxibustion stimulation of GV 4 region, a marked infrared radiant track along the central part of the back covering the GV was observed in the healthy volunteer subjects, and the skin temperature levels of both GV 4 and its bilateral control points were significantly increased (P < 0.001). The skin temperature of GV 4 was remarkably higher than those of both left and right control points (P < 0.001). After moxibustion stimulation, the average skin temperature levels of GV 9, GV 7, GV 4 and GV 3 were all up-regulated in comparison with pre-moxibustion (P < 0.01). CONCLUSION: Moxibustion stimulation of Mingmen (GV 4) can induce an infrared radiant track along the central part of the back covering the GV and raise skin temperature of acupoint areas of the GV.

[Simultaneous determination of thirteen components in Polygoni Multifori Caulis by UPLC-MS/MS].

A comprehensive analytical method based on UPLC-MS/MS was developed for the simultaneous determination of thirteen components including three stilbenes (stilbeneglucoside, polydatin, resveratrol), four anthraquinones (emodin, physcion, emodin-8-ß-D-glucopyranoside, aloe-emodin), five flavonoids (epicatechin, rutin, hyperoside, astragalin,quercetin) and one phenolic acid (gallic acid) in Polygoni Multifori Caulis.The separation was carried out on a Waters BEH C18 column（2.1 mm×100 mm,1.7 µm）with gradient elution of acetonitrile-water (0.1% acetic acid) at a flow rate of 0.25 mL•min⁻¹, and column temperature was 35 ℃. The target compounds were analyzed by multiple reaction monitoring (MRM) mode. TOPSIS analysis ware performed to evaluate the samples from different areas and commercial herbs according to the contents of thirteen components. The correlation coefficients of all the calibration curves were higher than 0.991 5. The average recoveries ranged from 95.24% to 102.3%, and the relative standard deviations were less than 5%. The result of TOPSIS analysis showed that the comprehensive quality of Polygoni Multifori Caulis sample from Guangzhou was better. The developed method with good repeatability and accuracy was suitable for the simultaneous determination of multiple functional substances, which provided a new basis for the comprehensive assessment and overall control of the quality of Polygoni Multifori Caulis.

Dynamic accumulation analysis on bioactive constituents of Polygonum multiflorum in different collection periods.

To study the dynamic change law of bioactive constituents from Polygonum multiflorum, and to explore the optimal harvest period of P. multiflorum. Determination of stilhene glucoside, anthraquinones and catechin from P. multiflorum in different harvest times by MEKC-DAD, and principal component analysis (PCA) was used to comprehensive evaluation for bioactive constituents. There are obvious differences among the contents of active ingredients in various collecting periods samples, the content of stilbene glucoside was the highest in November, the total content of combined anthraquinone was the highest in November and December, the content of catechin was the highest in September. The comprehensive evaluation index obtained with principal component analysis showed that the sample collected in November is significantly higher than those with other samples. The optimal harvest period of P. multiflorum is November.

[QTRAP LC-MS/MS Analytical Study on Nucleosides and Nucleobases of Pseudostellariae Radix Cultivated in Different Idioplasm Resources].

OBJECTIVE: To analyze nucleosides and nucleobases of Pseudostellariae Radix cultivated in different idibplasni resources and to compare the differences. METHODS: QTRAP LC-MS/MS method was applied for the analysis of 13 kinds of nucleosides and nucleobases in Pseudostellariae Radix and the data obtained was analyzed by SPSS 16. 0 software. RESULTS: There were some differences between Pseudostellariae Radix cultivated in different idioplasm resources. The highest amount of nucleosides and nucleobases was ZS2 which came from Zherong in Fujian Province. The total content of nucleosides and nucleobases in the sample from Shibing in Guizhou Province was the lowest. There was little difference between ZS1 (Zherong in Fujian Province) and XC(Xuancheng in Anhui Province). CONCLUSION: This study provides evidence for the influence of eco-environment on the metabolites of Pseudostellariae Radix.

[Effects of Electroacupuncture Stimulation of Zhongwan (CV 12) on Skin Microcirculatory Blood Perfusion Levels along the Conception Vessel in Yin-deficiency Constitution Volunteers].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation at Zhongwan (CV 12) on the skin microcirculatory blood flow along the Conception Vessel (CV) in yin-deficiency constitution volunteers. METHODS: A total of 16 yin-deficiency constitution volunteers (9 woman and 6 men) from 200 students of Fujian University of Traditional Chinese Medicine finished a body constitution questionnaire and showed a yin-deficiency constitution. Skin microcirculatory blood perfusion units (MBPU) at Danzhong (CV 17), Xiawan (CV 10) , and Qihai (CV 6) of the CV were measured by using a Laser Doppler Flowmetry (Peri Flux 5000) in normal constitution, before and after EA stimulation of Zhongwan (CV 12) for 20 min. RESULTS: Under normal condition, the cutaneous MBPU values were significantly lower in the Qihai (CV 6) than in both Danzhong (CV 17) and Xiawan (CV 10) (P < 0.01). After EA stimulation of CV 12 for 20 min, the MBPU values were significantly decreased in the CV 17 and CV 6 acupoint regions compared with pre-stimulation (P < 0.05). No significant changes of skin MBPU were found in Xiawan (CV 10) after EA stimulation of CV 12 (P > 0.05). CONCLUSION: EA at Zhongwan (CV 12) can reduce the skin MBPU values of CV 6 and CV 17 of the Conception Vessel in yin-deficiency constitution volunteers.

[Effects of different-frequency-electroacupuncture and moxibustion stimulation of "Neiguan" (PC 6) on arrhythmia in rabbits].

OBJECTIVE: To observe the effect of different-frequency-electroacupuncture (EA) and moxibustion stimulation on rabbits' arrhythmias. METHODS: Newzealand rabbits were randomly divided into arrhythmia model (n = 18), EA-3 Hz (n = 19), EA-20 Hz (n = 19), EA-100 Hz (n = 18) and moxibustion (n = 18) groups. The arrhythmia model was induced by intravenous injection of 25 µg/kg aconitine. EA (3 Hz, 20 Hz or 100 Hz, 3.5-4 V) or moxibustion was applied to bilateral "Neiguan" (PC 6) for 20 min and electrocardiogram (ECG) of the standard limb lead II was recorded by using a Biosignal Processing Instrument. The concentration of serum hypersensitive C reactive protein (hs-CRP) content was detected by using ELISA method. RESULTS: Following EA intervention, the duration of aconitine-induced arrhythmia was significantly shorter in both EA-20 Hz and EA-100 Hz groups than in the model group (P < 0.05, P < 0.01), and the serum hs-CRP level in the EA-20 Hz group was significantly lower than that of the model group ( P < 0.05). No significant changes were found in the duration of arrhythmia in both EA-3 Hz and moxibustion groups, and in the serum hs-CRP level in the EA-3 Hz, EA-100.Hz and the moxibustion groups (P > 0. 05). CONCLUSION: EA stimulation of PC 6 at 20 Hz and 100 Hz has an anti-arrhythmic effect in aconitine-induced arrhythmia rabbits.

[Dynamic Changes of Nucleosides and Nucleobases in Different Harvest Periods of Polygoni Multiflori Radix by UPLC-QTRAP-MS/MS].

OBJECTIVE: To analyze the dynamic changes of nucleosides and nucleobases in Polygoni Multiflori Radix harvested in different periods. METHODS: UPLC-QTRAP-MS/MS method was applied for the analysis of nine kinds of nucleosides and nucleobases in Polygoni Multiflori Radix. RESULTS: The content of uridine, adenine, guanosine and cytidine was higher in Polygoni Multiflori Radix harvested in different periods and assumed some difference. The trends of nucleosides and nucleobases from Polygoni Multiflori Radix according to the peak valley shape changed. The highest contents of them were in December. CONCLUSION: The accumulation of nucleosides and nucleobases in Polygoni Multiflori Radix is closely related to its growth cycle. It is found to be basically the same as that obtained when the herb is collected during the conventional collecting time.