ABSTRACT
BACKGROUND: Danggui Sini decoction (DSD), a traditional Chinese medicine formula, has the function of nourishing blood, warming meridians, and unblocking collaterals. Our clinical and animal studies had shown that DSD can effectively protect against oxaliplatin (OXA)-induced peripheral neuropathy (OIPN), but the detailed mechanisms remain uncertain. Multiple studies have confirmed that gut microbiota plays a crucial role in the development of OIPN. In this study, the potential mechanism of protective effect of DSD against OIPN by regulating gut microbiota was investigated. METHODS: The neuroprotective effects of DSD against OIPN were examined on a rat model of OIPN by determining mechanical allodynia, biological features of dorsal root ganglia (DRG) as well as proinflammatory indicators. Gut microbiota dysbiosis was characterized using 16S rDNA gene sequencing and metabolism disorders were evaluated using untargeted and targeted metabolomics. Moreover the gut microbiota mediated mechanisms were validated by antibiotic intervention and fecal microbiota transplantation. RESULTS: DSD treatment significantly alleviated OIPN symptoms by relieving mechanical allodynia, preserving DRG integrity and reducing proinflammatory indicators lipopolysaccharide (LPS), IL-6 and TNF-α. Besides, DSD restored OXA induced intestinal barrier disruption, gut microbiota dysbiosis as well as systemic metabolic disorders. Correlation analysis revealed that DSD increased bacterial genera such as Faecalibaculum, Allobaculum, Dubosiella and Rhodospirillales_unclassified were closely associated with neuroinflammation related metabolites, including positively with short-chain fatty acids (SCFAs) and sphingomyelin (d18:1/16:0), and negatively with pi-methylimidazoleacetic acid, L-glutamine and homovanillic acid. Meanwhile, antibiotic intervention apparently relieved OIPN symptoms. Furthermore, fecal microbiota transplantation further confirmed the mediated effects of gut microbiota. CONCLUSION: DSD alleviates OIPN by regulating gut microbiota and potentially relieving neuroinflammation related metabolic disorder.
ABSTRACT
Four previously undescribed angucyclinones umezawaones A-D (1-4) were isolated from the liquid cultures of Umezawaea beigongshangensis. Their structures were determined by spectroscopic analyses, single crystal X-ray diffraction, quantum chemical 13C NMR and electronic circular dichroism calculations. All compounds displayed strong inhibitory activities against indoleamine 2,3-dioxygenase and tryptophan-2,3-dioxygenase in enzymatic assay, especially compound 2.
Subject(s)
Actinobacteria , Tryptophan Oxygenase , Tryptophan Oxygenase/chemistry , Tryptophan Oxygenase/metabolism , Angucyclines and Angucyclinones , Actinomyces/metabolism , Indoleamine-Pyrrole 2,3,-Dioxygenase , Molecular StructureABSTRACT
Six new compounds, including two depsidones garciculendepsidones A and B (1 and 2), one prenylated xanthone garciculenxanthone (3) and three dimeric xanthones bigarciculenxanthones A-C (4-6), were isolated from the twigs and leaves of Garcinia esculenta Y. H. Li. Their structures were elucidated based on comprehensive analyses of spectral data, including HRESIMS, 1D and 2D NMR, and ECD calculation. All the isolates were tested for their cytotoxicity against five human cancer cell lines (myeloid leukemia HL-60, lung cancer A-549 cells, hepatocellular carcinoma SMMC-7721, breast cancer MDA-MB-231 and colon cancer SW480), among them, compounds 3-5 displayed cytotoxic potential, especially garciculenxanthone (3) had the lowest IC50 value of 8.2 µm for lung cancer A-549 cells.
Subject(s)
Antineoplastic Agents, Phytogenic , Antineoplastic Agents , Depsides , Garcinia , Lactones , Lung Neoplasms , Xanthones , Humans , Molecular Structure , Antineoplastic Agents, Phytogenic/pharmacology , Antineoplastic Agents, Phytogenic/chemistry , Garcinia/chemistry , Xanthones/pharmacology , Xanthones/chemistry , Lung Neoplasms/drug therapyABSTRACT
Microorganisms play an important role in regulating flavor compounds in rice wine, whereas we often don't understand how did they affect flavor compounds. Here, the relations between flavor compounds and microbial community ecological succession were investigated by monitoring flavor compounds and microbial community throughout the fermentation stage of rice wine. The composition of microbial community showed a dynamic change, but 13 dominant bacterial genera and 4 dominant fungal genera were detected throughout the fermentation stages. Saccharomyces presented a strong negative correlation with fungi genera but had positive associations with bacteria genera. Similarly, flavor compounds in rice wine were also showed the dynamic change, and 112 volatile compounds and 17 free amino acids were identified in the whole stages. The alcohol-ester ratio was decreased in the LTF stage, indicating that low temperature boosts ester formation. The potential correlation between flavor compounds and microbial community indicated that Delftia, Chryseobacterium, Rhizopus and Wickerhamomyces were the core functional microorganisms in rice wine. These findings clarified the correlation between changes in flavor compounds and in microbial community in the liquid fermentation of rice wine, and these results have some reference value for the quality improvement and technological optimization in liquid fermentation of rice wine.
Subject(s)
Microbiota , Wine , Fermentation , Dietary Supplements , EstersABSTRACT
To investigate the mechanism by which Cangxi Tongbi Capsules promote chondrocyte autophagy to inhibit knee osteoarthritis(KOA) progression by regulating the circRNA_0008365/miR-1271/p38 mitogen-activated protein kinase(MAPK) pathway. The cell and animal models of KOA were established and intervened with Cangxi Tongbi Capsules, si-circRNA_0008365, si-NC, and Cangxi Tongbi Capsules combined with si-circRNA_0008365. Flow cytometry and transmission electron microscopy were employed to determine the level of apoptosis and observe autophagosomes, respectively. Western blot was employed to reveal the changes in the protein levels of microtubule-associated protein light chain 3(LC3)â ¡/â , Beclin-1, selective autophagy junction protein p62/sequestosome 1, collagen â ¡, a disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTS-5), and p38 MAPK. The mRNA levels of circRNA_0008365, miR-1271, collagen â ¡, and ADAMTS-5 were determined by qRT-PCR. Hematoxylin-eosin staining was employed to reveal the pathological changes of the cartilage tissue of the knee, and enzyme-linked immunosorbent assay to measure the levels of interleukin-1ß(IL-1ß) and tumor necrosis factor-alpha(TNF-α). The chondrocytes treated with IL-1ß showed down-regulated expression of circRNA_0008365, up-regulated expression of miR-1271 and p38 MAPK, lowered autophagy level, increased apoptosis rate, and accelerated catabolism of extracellular matrix. The intervention with Cangxi Tongbi Capsules up-regulated the expression of circRNA_0008365, down-regulated the expression of miR-1271 and p38 MAPK, increased the autophagy level, decreased the apoptosis rate, and weakened the catabolism of extracellular matrix. However, the effect of Cangxi Tongbi Capsules was suppressed after interfering with circRNA_0008365. The in vivo experiments showed that Cangxi Tongbi Capsules dose-dependently inhibited the p38 MAPK pathway, enhanced chondrocyte autophagy, and mitigated articular cartilage damage and inflammatory response, thereby inhibiting the progression of KOA in rats. This study indicated that Cangxi Tongbi Capsules promoted chondrocyte autophagy by regulating the circRNA_0008365/miR-1271/p38 MAPK pathway to inhibit the development of KOA.
Subject(s)
MicroRNAs , Osteoarthritis, Knee , Rats , Animals , Chondrocytes , Osteoarthritis, Knee/metabolism , Osteoarthritis, Knee/pathology , RNA, Circular/genetics , RNA, Circular/metabolism , RNA, Circular/pharmacology , p38 Mitogen-Activated Protein Kinases/genetics , p38 Mitogen-Activated Protein Kinases/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , Apoptosis , Autophagy/genetics , Collagen/metabolismABSTRACT
OBJECTIVE: To explore the mechanism of electroacupunctureï¼EAï¼ in improving learning-memory ability in Alzheimer's disease ï¼ADï¼ mice from the perspective of endosomal-lysosomal system. METHODS: Male APP/PS1 transgenic mice were randomly divided into model group and EA group ï¼n=10 in each groupï¼ and 10 male C57BL/6 wild mice were taken as the normal group. EA ï¼1 Hz/50 Hz, 1 mAï¼ was applied at bilateral "Yongquan"ï¼KI1ï¼ and acupuncture was applied at "Baihui" ï¼GV20ï¼ for 15 min. The mice of the model and normal groups were subjected to restriction with the same method as those of the EA group for 15 min. The treatment was conducted once every other day for 6 weeks. The spatial learning-memory ability ï¼shown by escape latency of place navigation test and the time of crossing the target platform and total swimming distance in the target quadrant in 1 min of spatial probe test ï¼ was detected by Morris water maze test. The immunoactivity of senile plaques ï¼SPï¼ in the hippocampus tissue was detected by immunohistochemistry. The ultrastructural characters of hippocampal neurons were observed by transmission electron microscope, and the expression levels of Ras-related protein 5 ï¼Rab5ï¼, Ras-related protein 7 ï¼Rab7ï¼ and cathepsin D ï¼CTSDï¼ in the hippocampus were detected by Western blot, separately. RESULTS: Compared with the normal group, the escape latency, SP immunoactivity, and protein expression levels of Rab5, Rab7 and CTSD were significantly increased ï¼P<0.05, P<0.01ï¼, while the number of crossing the original platform and the total swimming distance in the platform quadrant were considerably reduced ï¼P<0.05ï¼ in the model group. In contrast to the model group, the EA group had a marked decrease in the escape latency, SP immunoactivity, and protein expression levels of Rab5, Rab7 and CTSD ï¼P<0.05, P<0.01ï¼, and a striking increase in the number of crossing the original platform and the swimming distance in the platform quadrant ï¼P<0.05ï¼. Results of transmission electron microscope showed an accumulation of endosome, lysosome, and endolysosomes in the hippocampal neurons in the model group, which was evidently milder in the EA group. CONCLUSION: EA of GV20 and KI1 can improve the learning-memory ability of AD mice, which may be related to its function in reducing hippocampal Aß deposition and down-regulating endosomal-lysosomal system activity.
Subject(s)
Alzheimer Disease , Electroacupuncture , Male , Mice , Animals , Mice, Inbred C57BL , Mice, Transgenic , Alzheimer Disease/genetics , Alzheimer Disease/therapy , Endosomes , Lysosomes/genetics , Plaque, AmyloidABSTRACT
BACKGROUND: Hawk tea, a medicinal and edible plant, has been consumed for thousands of years in Southwest China. To date, no unified food safety standard for Hawk tea has been established, and systematic research on the quality of Hawk tea is lacking. OBJECTIVE: The aim of this study was to develop a comprehensive evaluation method for the quality of Hawk tea based on inclusions content, high-performance liquid chromatography (HPLC) fingerprinting combined with the quantitative analysis of multiple components with a single marker (QAMS) method. METHODS: The contents of total flavonoids, total phenols, total polysaccharides, and total protein were determined using the colorimetric method. An effective comprehensive evaluation method was established to classify the 16 batches of samples based on HPLC fingerprint analysis combined with similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA), partial least-squares discrimination analysis (PLS-DA), and the QAMS method. RESULTS: Flavonoids were the main chemical components of Hawk tea. The accuracy of the QAMS method was verified by comparing the calculated results with those of the external standard method (ESM). No significant differences were found between the two methods. Additionally, the fingerprint of Hawk tea was also established. CONCLUSION: The method established in this study can be used for the comprehensive quality evaluation of Hawk tea and can also provide a reference for the quality evaluation of other herbal medicines.
Subject(s)
Drugs, Chinese Herbal , Drugs, Chinese Herbal/chemistry , Chromatography, High Pressure Liquid/methods , Quality Control , Flavonoids/analysis , Tea/chemistryABSTRACT
BACKGROUND: Jingfang granules (JFG), derived from JingFangBaiDu San (JFBDS), are a traditional herbal formulas used for the treatment of respiratory tract infections. They were initially prescribed to treat skin disease, such as psoriasis in Chinese Taiwan, but are not widely used for psoriasis treatment in mainland China because of the lack of anti-psoriasis mechanism research. PURPOSES: The present study was designed to evaluate the anti-psoriasis effect of JFG and reveal the correlated mechanisms of JFG in vivo and in vitro using network pharmacology, UPLC-Q-TOF-MS technology and molecular biotechnology methods. RESULTS: An imiquimod-induced psoriasis-like murine model was used to verify the anti-psoriasis effect in vivo, with inhibition of lymphocytosis and CD3+CD19+B cell proliferation in the peripheral blood and prevention of the activation of CD4+IL17+T cells and CD11c+ MHC â ¡+ dendritic cells (DCs) in the spleen. Network pharmacology analysis demonstrated that the targets of the active components were significantly enriched in pathways involved in cancer, inflammatory bowel disease and rheumatoid arthritis, which were closely related to cell proliferation and immune regulation. The drug-component-target networks and molecular docking analysis demonstrated the active ingredients to be luteolin, naringin and 6'-feruloylnodakenin, which had a good binding affinity to PPARγ, p38a MAPK and TNF-a. Finally, UPLC-Q-TOF-MS analysis to validate the active ingredients in drug-containing serum and in vitro experiments showed that JFG inhibited the maturation and activation of BMDCs via the p38a MAPK signaling pathway and translocation of the agonist PPARγ into the nuclei to reduce the activity of NF-κB/STAT3 inflammatory signaling pathway in keratinocytes. CONCLUSIONS: Our study demonstrated that JFG improved psoriasis by inhibiting the maturation and activation of BMDCs and proliferation and inflammation of keratinocytes, which may facilitate the applications of JFG in anti-psoriasis therapy in clinical settings.
Subject(s)
PPAR gamma , Psoriasis , Humans , Animals , Mice , PPAR gamma/metabolism , Molecular Docking Simulation , Aminoquinolines/adverse effects , Psoriasis/chemically induced , Psoriasis/drug therapy , Keratinocytes , Cell Division , Dendritic CellsABSTRACT
The human ether-a-go-go (hERG) potassium channel (Kv11.1) plays a critical role in mediating cardiac action potential. The blockade of this ion channel can potentially lead fatal disorder and/or long QT syndrome. Many drugs have been withdrawn because of their serious hERG-cardiotoxicity. It is crucial to assess the hERG blockade activity in the early stage of drug discovery. We are particularly interested in the hERG-cardiotoxicity of compounds collected in the DrugBank database considering that many DrugBank compounds have been approved for therapeutic treatments or have high potential to become drugs. Machine learning-based in silico tools offer a rapid and economical platform to virtually screen DrugBank compounds. We design accurate and robust classifiers for blockers/non-blockers and then build regressors to quantitatively analyze the binding potency of the DrugBank compounds on the hERG channel. Molecular sequences are embedded with two natural language processing (NLP) methods, namely, autoencoder and transformer. Complementary three-dimensional (3D) molecular structures are embedded with two advanced mathematical approaches, i.e., topological Laplacians and algebraic graphs. With our state-of-the-art tools, we reveal that 227 out of the 8641 DrugBank compounds are potential hERG blockers, suggesting serious drug safety problems. Our predictions provide guidance for the further experimental interrogation of DrugBank compounds' hERG-cardiotoxicity.
Subject(s)
Cardiotoxicity , Ether-A-Go-Go Potassium Channels , Humans , Ether-A-Go-Go Potassium Channels/chemistry , Ether-A-Go-Go Potassium Channels/metabolism , Ether , Potassium Channel Blockers/pharmacology , Potassium Channel Blockers/chemistry , Machine Learning , Ethyl Ethers , EthersABSTRACT
OBJECTIVE@#To compare the effect of electroacupuncture at Tinghui (GB 2) and Benshen (GB 13) combined with routine rehabilitation training and simple routine rehabilitation training on abnormal auditory response in children with autism spectrum disorder (ASD).@*METHODS@#A total of 70 children with ASD were randomly divided into an observation group and a control group, 35 cases in each group. The control group was given routine rehabilitation training. On the basis of the treatment in the control group, the observation group was given electroacupuncture at Tinghui (GB 2) and Benshen (GB 13), disperse-dense wave, frequency in 2 Hz/10 Hz, 20 min each time, and 1 day off after 6 days of treatment. Both groups were treated for 12 weeks. The incidence of abnormal auditory response, the scores of childhood autism rating scale (CARS) and autism behavior checklist (ABC) were compared between the two groups before and after treatment.@*RESULTS@#After treatment, the total incidences of abnormal auditory response in the two groups and the incidences of dull and hypersensitive auditory response in the observation group were lower than those before treatment (P<0.05). The total incidence of abnormal auditory response and the incidence of hypersensitive auditory response in the observation group were lower than those in the control group (P<0.05). After treatment, CARS and ABC scores of both groups were lower than those before treatment (P<0.05), and those in the observation group were lower than the control group (P<0.05).@*CONCLUSION@#Electroacupuncture at Tinghui (GB 2) and Benshen (GB 13) combined with routine rehabilitation training can reduce the incidence of abnormal auditory response and improve the core symptoms in children with ASD, and the clinical effect is better than the simple routine rehabilitation training.
Subject(s)
Humans , Child , Electroacupuncture , Autism Spectrum Disorder/therapy , Acupuncture Points , Medicine , PainABSTRACT
OBJECTIVE@#To observe the clinical efficacy of acupuncture for prevention of moderate to severe seasonal allergic rhinitis.@*METHODS@#A total of 105 patients with moderate to severe seasonal allergic rhinitis were randomly divided into an observation group (53 cases, 3 cases dropped off) and a control group (52 cases, 4 cases dropped off). The patients in the observation group were treated with acupuncture at Yintang (GV 24+), Yingxiang (LI 20), Hegu (LI 4), Zusanli (ST 36), Fengchi (GB 20), Feishu (BL 13), etc. 4 weeks before the seizure period, once every other day, 3 times a week for 4 weeks. The patients in the control group were not given any intervention before the seizure period. Emergency drugs can be given appropriately during the seizure period in both groups. After seizure period, the seizure rate was recorded in the two groups; before treatment and on week 1, 2, 4, 6 of seizure period after treatment, the rhinoconjunctivitis quality of life questionnaire (RQLQ) score and total nasal symptom score (TNSS) were observed in the two groups; the rescue medication score (RMS) was recorded on week 1-6 of seizure period in the two groups.@*RESULTS@#The seizure rate of the observation group was 84.0% (42/50), which was lower than 100.0% (48/48) in the control group (P<0.05). After treatment, the scores of RQLQ and TNSS at each time point of seizure period were decreased compared with before treatment in the observation group (P<0.01), which were lower than the control group (P<0.01). The RMS score at each time point of seizure period in the observation group was lower than the control group (P<0.05, P<0.01).@*CONCLUSION@#Acupuncture can reduce the incidence of moderate to severe seasonal allergic rhinitis, relieve the symptoms, improve the quality of life and reduce the use of emergency drugs.
Subject(s)
Humans , Rhinitis, Allergic, Seasonal , Rhinitis, Allergic/therapy , Quality of Life , Acupuncture Therapy , Acupuncture Points , Treatment Outcome , SeizuresABSTRACT
OBJECTIVE@#To observe the effect of electroacupuncture (EA) on liver protein kinase B (Akt)/forkhead box transcription factor 1 (FoxO1) signaling pathway in Zucker diabetic fatty (ZDF) rats, and to explore the possible mechanism of EA on improving liver insulin resistance of type 2 diabetes mellitus.@*METHODS@#Twelve male 2-month-old ZDF rats were fed with high-fat diet for 4 weeks to establish diabetes model. After modeling, the rats were randomly divided into a model group and an EA group, with 6 rats in each group. In addition, six male Zucker lean (ZL) rats were used as the blank group. The rats in the EA group were treated with EA at bilateral "Zusanli" (ST 36), "Sanyinjiao" (SP 6), "Weiwanxiashu" (EX-B 3), and "Pishu" (BL 20). The ipsilateral "Zusanli" (ST 36) and "Weiwanxiashu" (EX-B 3) were connected to EA device, continuous wave, frequency of 15 Hz, 20 min each time, once a day, six times a week, for a total of 4 weeks. The fasting blood glucose (FBG) in each group was compared before modeling, before intervention and after intervention; the serum levels of insulin (INS) and C-peptide were measured by radioimmunoassay method, and the insulin resistance index (HOMA-IR) was calculated; HE staining method was used to observe the liver tissue morphology; Western blot method was used to detect the protein expression of Akt, FoxO1 and phosphoenolpyruvate carboxykinase (PEPCK) in the liver.@*RESULTS@#Before intervention, compared with the blank group, FBG was increased in the model group and the EA group (P<0.01); after intervention, compared with the model group, FBG in the EA group was decreased (P<0.01). Compared with the blank group, the serum levels of INS and C-peptide, HOMA-IR, and the protein expression of hepatic FoxO1 and PEPCK were increased (P<0.01), while the protein expression of hepatic Akt was decreased (P<0.01) in the model group. Compared with the model group, the serum levels of INS and C-peptide, HOMA-IR, and the protein expression of hepatic FoxO1 and PEPCK were decreased (P<0.01), while the protein expression of hepatic Akt was increased (P<0.01) in the EA group. In the model group, the hepatocytes were structurally disordered and randomly arranged, with a large number of lipid vacuoles in the cytoplasm. In the EA group, the morphology of hepatocytes tended to be normal and lipid vacuoles were decreased.@*CONCLUSION@#EA could reduce FBG and HOMA-IR in ZDF rats, improve liver insulin resistance, which may be related to regulating Akt/FoxO1 signaling pathway.
Subject(s)
Male , Animals , Rats , Rats, Zucker , Proto-Oncogene Proteins c-akt/genetics , Diabetes Mellitus, Type 2/therapy , Insulin Resistance , C-Peptide , Electroacupuncture , Liver , Signal Transduction , Insulin , LipidsABSTRACT
This study aimed to explore the anti-inflammatory material basis and molecular mechanism of Artemisia stolonifera based on the analysis of the chemical components in different extracted fractions of A. stolonifera and their antioxidant and anti-inflammatory effects in combination with network pharmacology and molecular docking. Thirty-two chemical components were identified from A. stolonifera by ultra-performance liquid chromatography coupled to tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Among them, there were 7, 21 and 22 compounds in water, n-butanol and ethyl acetate fractions, respectively. The antio-xidant capacity of different extracted fractions was evaluated by measuring their scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) free radicals and total antioxidant capacity [ferric reducing antioxidant power(FRAP) assay]. The inflammatory model of RAW264.7 cells was induced by lipopolysaccharide(LPS), and the levels of nitrite oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) in the supernatant and the mRNA expression of related inflammatory factors in cells were used to evaluate the anti-inflammatory effects. The results revealed that ethyl acetate fraction of A. stolonifera was the optimal antioxidant and anti-inflammatory fraction. By network pharmacology, it was found that flavonoids such as rhamnazin, eupatilin, jaceosidin, luteolin and nepetin could act on key targets such as TNF, serine/threonine protein kinase 1(AKT1), tumor protein p53(TP53), caspase-3(CASP3) and epidermal growth factor receptor(EGFR), and regulate the phosphatidylinositol-3-kinase-protein kinase B(PI3K-AKT) and mitogen-activated protein kinase(MAPK) signaling pathways to exert the anti-inflammatory effects. Molecular docking further indicated excellent binding properties between the above core components and core targets. This study preliminarily clarified the anti-inflammatory material basis and mechanism of ethyl acetate fraction of A. stolonifera, providing a basis for the follow-up clinical application of A. stolonifera and drug development.
Subject(s)
Antioxidants/chemistry , Molecular Docking Simulation , Artemisia , Network Pharmacology , Phosphatidylinositol 3-Kinases , Anti-Inflammatory Agents/chemistry , Drugs, Chinese Herbal/pharmacology , Interleukin-6ABSTRACT
OBJECTIVE: To investigate the effect of electroacupuncture (EA) on the expression of ß-amyloid (Aß) and autophagy-related proteins in hippocampal cells of Alzheimer's disease (AD) model mice, so as to explore its underlying mechanisms. METHODS: Eighteen male APP/PS1 transgenic mice (6 months old) were randomly divided into model and EA groups, with 9 mice in each group. Nine male C57BL/6 wild-type mice of the same age were chosen as the normal group. Mice in the EA group were treated with acupuncture on "Baihui" (GV20) and EA (1 Hz/50 Hz, 1 mA) on bilateral "Yongquan" (KI1), once every other day, 20 min each time for a total of 21 times. After the interventions, the spatial learning and memory ability were observed by Morris water maze test. The autophagy-related pathological changes in hippocampus were observed by transmission electron microscopy. The expressions of microtublue associated protein 1 light chain 3 (LC3) and Aß in hippocampus were observed by immunofluorescence and the expression levels of LC3 and p62 proteins were detected by Western blot. RESULTS: Compared with the normal group, the escape latency was prolonged (P<0.01), the residence time in the original quadrant platform was shor-tened (P<0.05), the positive expressions of LC3 and Aß, the expression levels of LC3â ¡ and p62 proteins, and the ratio of LC3â ¡/LC3â proteins in hippocampus were increased (P<0.01, P<0.05) in the model group. Compared with the model group, the escape latency was shortened (P<0.05), the residence time in the original quadrant platform was prolonged (P<0.05), the positive expressions of LC3 and Aß, the expression levels of LC3â ¡ and p62 proteins, and the ratio of LC3â ¡/LC3â proteins in hippocampus were decreased (P<0.05) in the EA group. The transmission electron microscopy showed that the structure of neurons was normal in the normal group, a large number of autolysosomes and autophagosomes existed in hip-pocampal nerve cells in the model group, and only a small number of autophagosomes were observed in the EA group. CONCLUSION: EA can reduce the expression levels of autophagy-related proteins LC3 and p62 in APP/PS1 transgenic mice, improve the hip-pocampal autophagy state, reduce intracellular Aß aggregation, and thus improve the learning and memory ability.
Subject(s)
Alzheimer Disease , Electroacupuncture , Mice , Male , Animals , Alzheimer Disease/genetics , Alzheimer Disease/therapy , Amyloid beta-Peptides/genetics , Amyloid beta-Peptides/metabolism , Autophagy-Related Proteins/metabolism , Mice, Inbred C57BL , Hippocampus/metabolism , Mice, Transgenic , Spatial LearningABSTRACT
OBJECTIVE: To observe the clinical effect of acupotomy combined with warm needling on cervical spondylotic radiculopathy (CSR) of qi and blood stagnation syndrome. METHODS: A total of 90 CSR patients were randomly divided into an acupotomy group, a warm needling group and a combined treatment group, with 30 cases in each group. The patients in the acupotomy group were treated with acupotomy, once every 7 days, consecutively for 3 times. The patients in the warm needling group received warm needling, once daily, at the interval of 2 days after consecutive treatments for 5 days, 7 days as one session of treatment and 3 consecutive sessions were required. The patients in the combined treatment group were treated with acupotomy and warm needling, and the methods and the treatment session were same as the the previous two groups. Before and after the treatment, the pain rating index (PRI) of McGill pain questionnaire (MPQ) and the 20-point scale of CSR developed by Yasuhisa Tanaka (CSR20) were adopted in the assessment. The changes of clinical symptoms and functions of patients were observed and the clinical efficacy was assessed in each group. RESULTS: After the treatment, the PRI score was decreased (P<0.05) and the CSR20 score was increased (P<0.05) in the 3 treatment groups when compared with those before the treatment. After the treatment, compared with the acupotomy group and the warm needling group, the PRI score was decreased (P<0.05) and the CSR20 score was increased (P<0.05) in the combined treatment group. The total effective rate was 83.3% (25/30) in the acupotomy group, 76.7% (23/30) in the warm needling group and 93.3% (28/30) in the combined treatment group. The total effective rate in the combined treatment group was higher than those in the acupotomy group and the warm needling group (P<0.05). CONCLUSION: The combined treatment with acupotomy and warm needling may obviously improve the clinical symptoms and physical signs, e.g. pain and numbness in the patients with CSR of qi and blood stagnation syndrome. Its efficacy is remarkably higher than that of the simple application of acupotomy or warm needling.
Subject(s)
Acupuncture Therapy , Radiculopathy , Spondylosis , Humans , Radiculopathy/therapy , Qi , Spondylosis/therapy , Acupuncture Therapy/methods , Treatment Outcome , Syndrome , PainABSTRACT
L-lysine is a crucial nutrient for both humans and animals, and its main commercial use is as a supplement in animal feed to promote chicken and other animal growth. Fluorescence biosensors based on the transcriptional regulator have been developed for high-throughput screening of L-lysine producers. However, due to its inability to specifically detect lysine, this fluorescent biosensor cannot be employed to screen high-yielding strains. Here, we present a novel technique for observing L-lysine concentrations within individual Corynebacterium glutamicum cells. The transcriptional regulator LysG and its binding site, as well as the phytoene desaturase that catalyzes the synthesis of the red pigment, make up the functional core of the biosensor. The lysine-sensitive mutant LysG(E123Y, E125A), which improved the sensitivity of biosensors, was generated by site-directed saturation mutagenesis. In addition, we increased the lysine-induced chromogenic biosensor response to 320 mM by optimizing the L-lysine export mechanism and the pathway for the synthesis of lycopene precursors. The direct identification of producers with elevated L-lysine accumulation is thus made straightforward by colorimetric screening. Lys-8, a lysine producer with a maximum lysine titer of 316.2 mM, was sorted out based on the biosensor. The enzymatic colorimetric biosensor constructed here is a simple tool with great potential for the development of high-level lysine-producing C. glutamicum.
Subject(s)
Biosensing Techniques , Corynebacterium glutamicum , Biosensing Techniques/methods , Colorimetry , Corynebacterium glutamicum/metabolism , Humans , Lycopene/metabolism , Lysine/metabolismABSTRACT
OBJECTIVE: To observe the clinical efficacy of balance acupotomy combined with warm needling in treatment of cervical spondylopathy of vertebral artery type (CSA). METHODS: Ninety patients were randomly divided into a warm needling group, a balance acupotomy group and a combined treatment group, 30 cases in each one. In the warm needling group, warm needling was given, once daily, for 5 days consecutively, with the interval of 2 days every week, and the treatment was conducted for 3 weeks. In the combined treatment group, on the base of the treatment as the warm needling group, the balance acupotomy was exerted, for consecutive 3 weeks. In the balance acupotomy group, the balance acupotomy was adopted, once a week, for 3 weeks consecutively. Clinical efficacy, the mean blood flow velocity of left vertebral artery (LVA), right vertebral artery (RVA) and basilar artery (BA), the pulse index (PI) and the resistance index (RI) were observed in 3 groups separately. The score of vertigo symptom and function was compared before and after treatment in 3 groups. RESULTS: In comparison with before treatment, the score of evaluation scale for cervical vertigo (ESCV) and the mean blood flow velocity of LVA, RVA and BA were all increased (P<0.05). while PI and RI reduced (P<0.05) in each group after treatment. In comparison with those in the warm needling group and the balance acupotomy group, ESCV score and the mean blood flow velocity of LVA, RVA and BA were increased (P<0.05), while PI and RI decreased (P<0.05) in the combined treatment group after treatment. Compared with the balance acupotomy group, the mean blood flow velocity of LVA, RVA and BA was increased (P<0.05), and PI and RI reduced (P<0.05) in the warm needling group after treatment. The total effective rate was 73.3% (22/30) in the warm needling group, 70.0% (21/30) in the balance acupotomy group and 93.3% (28/30) in the combined treatment group respectively. The total effective rate in the combined treatment group was higher than that either in the warm needling group or in the balance acupotomy group (P<0.05). CONCLUSION: Balance acupotomy combined with warm needling may restore the mechanical equilibrium state of the neck, and effectively improve the change of vertebral-basilar artery blood flow and relieve vertigo symptoms in patient with of cervical spondylosis of vertebral artery type.
Subject(s)
Acupuncture Therapy , Spondylosis , Basilar Artery , Dizziness , Humans , Spondylosis/therapy , Treatment Outcome , Vertebral Artery/diagnostic imaging , Vertigo/therapyABSTRACT
OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expression of Iba-1, complement C1q and CD68 in hippocampus of SAMP8 mice, so as to explore its mechanisms underlying improvement of Alzheimer's disease (AD). METHODS: Twenty-four male SAMP8 mice were randomly and equally divided into model and EA groups, and 12 SAMR1 mice were used as the control group. EA (2 Hz, 1.5-2.0 mA) was applied to "Baihui" (GV20), "Dazhui"(GV14) and "Shen-shu"(BL23) for 20 min once daily in the EA group, each course of treatment was 8 days, with an interval of 2 days between two courses, and the mice were treated for 3 courses. Morris water maze test was performed to assess the learning-memory ability of mice. The positive expression levels of Iba-1 and CD68 proteins in the hippocampus CA1 region were detected by immunohistochemistry. The mRNA and protein expression levels of Iba-1,C1q and CD68 in the hippocampus were detected by real-time PCR and Western blot, separately. RESULTS: Compared with the control group, the average escape latency of Morris water maze test was prolonged in the model group (P<0.01), duration of swimming in the original platform quadrant and the number of original platform crossing were significantly shorter and decreased respectively (P<0.01). Compared with the model group, the average escape latency in the EA group was shortened (P<0.05, P<0.01), the duration of swimming in the original platform quadrant and the number of original platform crossing were significantly prolonged and increased (P<0.01). The immunoactivity of Iba-1 and CD68 in hippocampal CA1 region, and mRNA and protein expression levels of hippocampal Iba-1,C1q and CD68 were significantly up-regulated in the model group in contrast to the control group (P<0.01, P<0.05), and obviously down-regulated except the mRNA expression level of hippocampal Iba-1 in the EA group relevant to the model group (P<0.01, P<0.05). CONCLUSION: EA can improve the learning and memory ability of SAMP8 mice, which may be associated with its effect in inhibiting of complement C1q-dependent microglial phagocytosis in the hippocampus.
Subject(s)
Electroacupuncture , Animals , Complement C1q/genetics , Complement C1q/metabolism , Hippocampus/metabolism , Male , Memory , Mice , Microglia/metabolism , Phagocytosis , RNA, Messenger/metabolismABSTRACT
Mulberry (Morus alba L.) fruit is a fruit with nutritional and medicinal value. It is widely cultivated in different regions of China, which may result in differences in its chemical composition. In this research, 25 mulberry fruit samples from six provinces in China were investigated. The contents of anthocyanins were evaluated by high-performance liquid chromatography (HPLC). The contents of two main anthocyanins, cyanidin-3-O-glucoside (C3G) and cyanidin-3-O-rutinoside (C3R), ranged from 0.656 ± 0.006 mg/g to 4.367 ± 0.243 mg/g and from 0.226 ± 0.007 mg/g to 1.649 ± 0.013 mg/g, respectively. Additionally, the contents of total phenolic, total flavonoid, vitamin C, titratable acids, reducing sugars and antioxidant capacity (FRAP, DPPH, scavenging and hydroxyl radical scavenging activity) were also assessed. The results and principal component analysis showed that the Zhongsang 5801 variety from Sichuan, Dechang had the greatest health value with the highest active compound contents. Based on our analysis, the variety from Sichuan, Dechang is a high-quality plant source for mulberry fruit cultivation. This research provides a basis for the rational development and utilization of mulberry fruit resources in China.
Subject(s)
Anthocyanins , Morus , Anthocyanins/chemistry , Antioxidants/chemistry , Fruit/chemistry , Morus/chemistry , Plant Extracts/chemistryABSTRACT
Salvadora persica L. (Sp) has been widely used as folk medicine. This study aimed to identify and assess the significant phenolics' antioxidant activities in young (S1) and old (S2) leaves of Sp. Six flavonoids; isoquercitrin, kaempferol-3-neohesperidoside, myricetin-3-galactoside, apigenin-O-hexoside, isorhamnetin and isorhamnetin-3-neohesperidoside, were identified for the first time in Sp leaves using LC-ESI-QTOF MS/MS analysis. The flavonoid-rich fraction obtained after purification of S1 (S1TF7) has the highest TFC (358.88 ± 0.12 mg rutin equivalent/g) and TPC (180.82 ± 0.82 mg gallic acid equivalent/g) and exhibits significant (p < 0.05) free-radical scavenging activity in DPPH(IC50% = 14.64 ± 0.29 µg/mL) and ABTS (IC50% = 200.41 ± 1.85 µg/mL) assays which was similar to Butylated hydroxytoluene (BHT) as a positive control (IC50% =12.94 ± 0.10 µg/mL and IC50% = 190.47 ± 8.93 µg/mL, respectively). Also, S1TF7 inhibits Lipopolysaccharide (LPS)-stimulated ROS production in raw-cell-264.7 with no significant toxicity at 200 µg/mL (p > 0.05). The study concludes that S1TF7 can serve as a mean to prevent oxidative diseases, and it merits further pharmacological investigations.