The impact of electroacupuncture on anxiety-like behavior and gut microbiome in a mouse model of chronic restraint stress.

Introduction: Electroacupuncture (EA) is a beneficial physiotherapy approach for addressing neuropsychiatric disorders. Nevertheless, the impact of EA on the gut microbiome in relation to anxiety disorders remains poorly understood. Methods: To address this gap, we conducted a study using a chronic restraint stress (CRS) mouse model to investigate the anti-anxiety outcome of EA and its influence on gut microbiota. Our research involved behavioral tests and comprehensive sequencing of full-length 16S rRNA microbiomes. Results: Our findings revealed that CRS led to significant anxiety-like behaviors and an imbalance in the gut microbiota. Specifically, we identified 13 species that exhibited changes associated with anxiety-like behaviors. Furthermore, EA partially alleviated both behaviors related to anxiety and the dysbiosis induced by CRS. Discussion: In summary, this study sheds light on the alterations in gut microbiota species resulting from CRS treatment and brings new light into the connection between EA's anti-anxiety effects and the gut microbiota.

SmMYB4 Is a R2R3-MYB Transcriptional Repressor Regulating the Biosynthesis of Phenolic Acids and Tanshinones in Salvia miltiorrhiza.

Salvia miltiorrhiza Bunge is one of the most famous traditional Chinese medicinal plants. The two most important classes of pharmaceutically relevant compounds in S. miltiorrhiza are phenolic acids and tanshinones. The MYB family of transcription factors may efficiently regulate the secondary metabolism in plants. In this study, a subgroup 4 R2R3MYB transcription factor gene, SmMYB4, was isolated from S. miltiorrhiza and functionally characterized using overexpression and a RNAi-mediated silencing. We achieved a total of six overexpressions and eight RNAi transgenic lines from the Agrobacterium leaf disc method. The content of the total phenolics, rosmarinic acid, and salvianolic acid B markedly decreased in the SmMYB4-overexpressing lines but increased in the SmMYB4-RNAi lines. The content of the total tanshinones, cryptotanshinone, and tanshinone IIA decreased in the SmMYB4-overexpressing transgenic lines but increased in the SmMYB4-RNAi lines. A gene expression analysis demonstrated that SmMYB4 negatively regulated the transcription of the critical enzyme genes involved in the phenolic acid and tanshinone biosynthesis. The genetic control of this transcriptional repressor may be used to improve the content of these bioactive compounds in the cultivated S. miltiorrhiza.

The efficacy of acupuncture on the sleep structure of patients with insomnia: A systematic review and meta-analysis.

This study aims to evaluate the efficacy of acupuncture on the sleep structure of patients with insomnia, so as to provide a valuable basis for the effectiveness of acupuncture in the treatment of insomnia. We conducted searches based on MeSH terms and free words in Cochrane Library, PubMed, Web of science, CKNI (China Knowledge Resource Integrated Database), WanFang Database, and Chongqing VIP Information from the inception of these database until 10 July 2020 for randomized controlled trials (RCTs) that investigated acupuncture treatment in patients with insomnia, and pertinent details of the results were saved. Comprehensive analysis showed that: (1) compared with the Western medicine groups, the acupuncture groups showed significant advantages in reducing the percentage of N1 sleep stage and N2 sleep stage, as well as increasing that of N3 sleep stage and REM sleep stage. However, no significant difference was found in increasing the effective rate, reducing total PSQI score, improving the total sleep time, reducing sleep latency, and improving sleep efficiency between the Western medicine groups and the acupuncture groups. (2) Compared with the sham acupuncture groups, the acupuncture treatment showed advantages in increasing the effective rate, reducing Pittsburgh Sleep Quality Index (PSQI) score, increasing the total sleep time, and improving sleep efficiency. However, no significant difference was observed between the sham acupuncture groups and the acupuncture groups with regard to reducing sleep latency, the percentage of N1 sleep stage and N2 sleep stage, as well as increasing that of N3 sleep stage and REM sleep stage.

Effect of acupuncture on sleep quality and neurological function in stroke patients with sleep apnea syndrome.

OBJECTIVE: To investigate the effect of acupuncture on sleep quality and neurological function in stroke patients with sleep apnea syndrome (SAS). METHODS: In this prospective study, a total of 88 stroke patients with SAS were randomized into two groups: observation group (44 cases; patients received the western medicine treatment combined with acupuncture) and control group (44 cases; patients were given western medicine treatment only). All patients in both groups were treated for three weeks. The clinical efficacy, sleep quality, apnea-hypopnea index (AHI), cognitive function, neuron-specific enolase (NSE) and S100 calcium binding protein ß (S100ß) levels before and after treatment were compared between patients in the two groups. RESULTS: Compared with those before treatment, the sleep latency, sleep duration, sleep efficiency and minimal oxygen saturation (SaO2min) increased, while the longest apnea time and AHI decreased in both groups after treatment. More significant changes were found in the observation group (all P<0.05). After treatment, the overall effective rate in the observation group was higher than that in the control group (P<0.05); serum levels of NSE and S100ß in both groups were lower than those before treatment, and the levels of the observation group were lower than those of the control group (all P<0.05); Montreal Cognitive Assessment (MoCA) scores in both groups were higher than those before treatment, and scores of the observation group were higher than those of the control group (all P<0.05). CONCLUSION: Western medicine treatment combined with acupuncture can significantly relieve the clinical symptoms of stroke patients with SAS and improve sleep quality and neurological function. Therefore, it is worthy of clinical application.

Neuroprotective effects of aucubin on hydrogen peroxide-induced toxicity in human neuroblastoma SH-SY5Y cells via the Nrf2/HO-1 pathway.

BACKGROUND: When redox balance is lost in the brain, oxidative stress can cause serious damage that leads to neuronal loss, in congruence with neurodegenerative diseases. Aucubin (AU) is an iridoid glycoside and that is one of the active constituents of Eucommia ulmoides, has many pharmacological effects such as anti-inflammation, anti-liver fibrosis, and anti-atherosclerosis. PURPOSE: The present study aimed to evaluate the inhibitory effects of AU on cell oxidative stress against hydrogen peroxide (H2O2)-induced injury in SH-SY5Y cells in vitro. METHODS: SH-SY5Y cells were simultaneously treated with AU and H2O2 for 24 h. Cell viability was measured by CCK-8. Additionally, mitochondrial membrane depolarization, reactive oxygen species (ROS) generation, and cell apoptosis were measured by flow cytometry. RESULTS: The results showed that AU can significantly increase the H2O2-induced cell viability and the mitochondrial membrane potential, decrease the ROS generation, malondialdehyde (MDA), and increase glutathione (GSH) contents and the superoxide dismutase (SOD) activity. We also found that H2O2 stimulated the production of nitric oxide (NO), which could be reduced by treatment with AU through inhibiting the inducible nitric oxide synthase (iNOS) protein expression. In H2O2-induced SH-SY5Y cells, the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) content and cell apoptosis were significantly reduced by AU treatment through nuclear factor E2-related factor 2/hemo oxygenase-1 (Nrf2/HO-1) activation, inhibiting the expression of p-NF-κB/NF-κB and down-regulating MAPK and Bcl-2/Bax pathways. CONCLUSION: These results indicate that AU can reduce inflammation and oxidative stress through the NF-κB, Nrf2/HO-1, and MAPK pathways.

Progress in Application of Population Pharmacokinetic and Pharmacodynamic Models in Research on Traditional Chinese Medicine / 中国实验方剂学杂志

In recent years， the role of quantitative pharmacological models in applicable population of drugs and dose optimization has been widely recognized. In order to improve the efficiency of clinical development and optimize clinical rational drug use， quantitative pharmacological models are being gradually introduced into the research of traditional Chinese medicine （TCM）. There are various types of quantitative pharmacological models， among which the following three models are commonly used：①Population pharmacokinetic （PPK） model， which is mainly used to explore the pharmacokinetic characteristics in different populations.②Pharmacokinetic-pharmacodynamic （PK-PD） model， which is used to reveal the internal relationship among dose， time and efficacy. ③PPK-PD model， which integrates both the characteristics of PPK model and PK-PD model. The paper summarizes the application of the above three models in TCM， and extracts the main ideas and methods of TCM model research， in order to provide reference for clinical research and rational use of TCM.

Engineering of ATP-Powered Photosensitizer for Targeted Recycling Activatable Imaging of MicroRNA and Controllable Cascade Amplification Photodynamic Therapy.

Owing to the low abundance of microRNAs (miRNAs) in living tumor cells, the development of intracellular cancer-relevant miRNA stimuli-activatable photosensitizers (PSs) for accurate imaging and efficient photodynamic therapy (PDT) of tumors in vivo is extremely challenging. Herein, we engineered a tumor targeting and intracellular trace miRNA-activatable nanophotosensitizer Y-motif/FA@HyNP on the basis of an endogenous ATP-powered strand-displacement cascade amplification strategy, which was prepared by assembly of a quencher BHQ2-labeled Y-motif DNA structure (containing ATP-binding aptamer and target miRNA-binding complementary sequence) on the surface of folate (FA) and amine-functionalized hybrid micellar nanoparticles. We showed that the fluorescence emissions at both 555 and 627 nm were effectively inhibited due to BHQ2 in Y-motif/FA@HyNPs, leading to negligible PDT efficacy. Once Y-motif/FA@HyNPs were selectively internalized into tumor cells via FA-receptor-mediated endocytosis, the intracellular trace target miRNA initiated the dissociation of the BHQ2-terminated sequences from Y-motif/FA@HyNPs by means of abundant endogenous ATP-powered strand-displacement reactions, causing remarkable fluorescence enhancement and cascade amplification PDT. The activated dual-color fluorescence emissions at 555 and 627 nm were feasible to achieve real-time, highly sensitive, and specific imaging of trace target miRNA in living tumor cells. With the guidance of excellent imaging in living mice, Y-motif/FA@HyNPs exhibited the precise and efficient PDT of tumors as well as insignificant side effects in vivo. This work revealed the great potential of using an integration of receptor-mediated cell uptake and target-triggered recycling cascade amplification strategy to design early cancer-relevant stimuli-activatable PSs for both fluorescence imaging and PDT ablation of tumors in vivo, which could effectively facilitate the timeliness and precision of early cancer diagnosis and therapy.

New tanshinone I derivatives S222 and S439 similarly inhibit topoisomerase I/II but reveal different p53-dependency in inducing G2/M arrest and apoptosis.

Tanshinone I (Tanshinone-1), a major active principle of the traditional Chinese medicine Salvia miltiorrhiza, possesses excellent anticancer properties, including inhibiting proliferation, angiogenesis and metastasis and overcoming multidrug resistance (MDR). However, its direct anticancer molecular target(s) remain unknown. Here we report that tanshinone-1 and its two new derivatives, S222 and S439, directly inhibit DNA topoisomerase I/II (Top1/2). With significantly improved water solubility, S222 and S439 displayed 12- and 14-times more potent proliferative inhibition than their parent tanshinone-1 in a panel of 15 cancer cell lines. Both retained tanshinone-1's anti-MDR and anti-angiogenesis properties and its capability to reduce the phosphorylation of Stat3 at Tyr705 with apparently enhanced efficacy and in these regards, S439 was also slightly more potent than S222. Both derivatives and tanshinone-1 directly inhibited Top1 and Top2 at molecular and cellular levels; the derivatives displayed similar potency but both were more potent than tanshinone-1. The inhibition of S222 and S439 on Top1 and Top2 was also more potent than that of the Top1 inhibitor hydroxylcamptothecin and the Top2 inhibitor etoposide, respectively. Consistently, tanshinone-1 and its derivatives induced DNA double-strand breaks, G2/M arrest and apoptosis. Unexpectedly, the derivatives demonstrated different p53-dependency in inducing both cell cycle arrest and apoptosis. S222 showed no obvious p53-dependency. In contrast, S439 induced more G2/M arrest in p53-proficient cells than in p53-deficient cells while its apoptotic induction was the opposite. However, their proliferative inhibition was independent of the p53 status. Due to their structures different from the known Top1, Top2 and dual Top1/2 inhibitors, our results indicate that tanshinone-1 and its derivatives are a new type of dual Top1/2 inhibitors.

EGFR­associated pathways involved in traditional Chinese medicine (TCM)­1­induced cell growth inhibition, autophagy and apoptosis in prostate cancer.

Traditional Chinese medicine (TCM) has the synergistic effect of the combination of a single ingredient and a monomer, and systemic and local therapeutic effects in cancer treatment, through which TCM is able to enhance the curative effect and reduce the side effects. The present study analyzed the effect of TCM­1 (an anti­cancer TCM) on prostate cancer (PCa) cell lines, and studied in detail the mechanism of cell death induced by TCM­1 in vitro and in vivo. From the present results, it was identified for the first time, to the best of our knowledge, that TCM­1 arrested the cell cycle at the G1 phase, decreased cell viability and increased nuclear rupture in a dose­dependent manner; these effects finally resulted in apoptosis in PCa cells. At the molecular level, the data demonstrated that TCM­1 competitively acted on epidermal growth factor receptor (EGFR) with EGF, and suppressed the auto­phosphorylation and activity of EGFR. Inhibition of EGFR further suppressed the downstream phosphatidylinositol 3­kinase (PI3K)/RAC­α serine/threonine­protein kinase (AKT) and RAF proto­oncogene serine/threonine­protein kinase/extracellular signal regulated kinase signaling pathways and resulted in a decrease in the phosphorylated­forkhead box protein O1 (at Ser256, Thr24 and Ser319) expression level, and induced cell growth inhibition and apoptosis by regulating the expression of apoptosis­and cell cycle­associated genes. In addition, TCM­1 markedly inhibited the PI3K/AKT/serine/threonine­protein kinase mTOR signaling pathway and induced cell autophagy by downregulating the phosphorylation of p70S6K and upregulating the levels of Beclin­1 and microtubule­associated protein light chain­3II. In vivo, the TCM­1­treated group exhibited a significant decrease in tumor volume compared with the negative control group in subcutaneous xenograft nude mice by inhibiting EGFR­associated signaling pathways. Therefore, the bio­functions of Chinese medicine TCM­1 in inducing PCa cell growth inhibition, autophagy and apoptosis suggested that TCM­1 may have clinical potential for the treatment of patients with PCa.

Traditional Chinese Medicine CFF-1 induced cell growth inhibition, autophagy, and apoptosis via inhibiting EGFR-related pathways in prostate cancer.

Traditional Chinese medicine (TCM) has a combined therapeutic result in cancer treatment by integrating holistic and local therapeutical effects, by which TCM can enhance the curative effect and reduce the side effect. In this study, we analyzed the effect of CFF-1 (alcohol extract from an anticancer compound Chinese medicine) on prostate cancer (PCa) cell lines and studied in detail the mechanism of cell death induced by CFF-1 in vitro and in vivo. From our data, we found for the first time that CFF-1 obviously arrested cell cycle in G1 phase, decreased cell viability and then increased nuclear rupture in a dose-dependent manner and finally resulted in apoptosis in prostate cancer cells. In molecular level, our data showed that CFF-1 induced inhibition of EGFR auto-phosphorylation and inactivation of EGFR. Disruption of EGFR activity in turn suppressed downstream PI3K/AKT and Raf/Erk signal pathways, resulted in the decrease of p-FOXO1 (Ser256) and regulated the expression of apoptosis-related and cycle-related genes. Moreover, CFF-1 markedly induced cell autophagy through inhibiting PI3K/AKT/mTOR pathway and then up-regulating Beclin-1 and LC-3II and down-regulating phosphorylation of p70S6K. In vivo, CFF-1-treated group exhibited a significant decrease in tumor volume compared with the negative control group in subcutaneous xenograft tumor in nude mice via inhibiting EGFR-related signal pathways. Thus, bio-functions of Chinese medicine CFF-1 in inducing PCa cell growth inhibition, autophagy, and apoptosis suggested that CFF-1 had the clinical potential to treat patients with prostate cancer.

[Effects of Cangfu Congxian Decoction on Oxidative Stress in Polycystic Ovary Syndrome Patients].

OBJECTIVE: To observe the effect of Cangfu Congxian Decoction (CCD) on oxidative stress in granulosa cells of polycystic ovary syndrome (PCOS) patients. METHODS: Forty PCOS patients underwent in vitro fertilization-embryo transfer (IVF-ET) were assigned to the treatment group and the control group 1 according to random digit table, 20 in each group. Patients in the treatment group took CCD (200 mL, once in the morning and once in the afternoon) 2 months before IVF-ET, while those in the control group 1 took no Chinese medical decoction. Recruited were another 20 patients undergoing IVF-ET for tubal factors (as the control group 2). The clinical effect of IVF-ET were observed, including oocyte retrieval number, 2 pronuclear (2PN) fertilization rate, good quality embryo rate, clinical pregnancy rate, and ovarian hyperstimulation syndrome (OHSS) induced transplantation cancel rate. The expression of relative oxygen species (ROS) in granulosa cells was detected using cell immunofluorescence combined with confocal microscopy and FCM. RESULTS: Compared with the control group 1, occyte retrieval number, 2PN fertilization rate, and good quality embryo rate increased in the control group 2 and the treatment group (P <0. 05). OHSS induced transplantation cancel rate decreased in the control group 2 (P < 0.05). Fluorescence intensity of ROS decreased in the treatment group and the control group 2, as compared with the control group 1 (P < 0.01). CONCLUSION: CCD increased good quality embryo rate by down-regulating the expression of ROS protein in ovarian granulosa cells, and correcting in vivo oxidative stress.

[Effects of Bushen Wenyang Huayu Recipe on Expressions of HIF-1α, PHD2, and VHL in Endometriosis Rats with Shen Yang Deficiency Blood Stasis Syndrome].

OBJECTIVE: To observe the effect of Bushen Wenyang Huayu Recipe (BWHR) on hypoxia inducible factor-1α (HIF-1α), proline hydroxylase2 (PHD2), von Hippel Lindau disease (VHL) suppressor gene expressions in endometriosis (EM) rats with Shen yang deficiency blood stasis syndrome (SYDBSS), and to explore the pathogenesis of EM and the mechanism of BWHR for treating EM. METHODS: Totally 50 SD rats were randomly divided into five groups, i.e., the blank control group, the sham-operation group, the model group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 10 in each group. Rats in the blank control group and the sham-operation group were fed routinely. Rats in the rest 3 groups received 30-day "extended refrigerator freezing and ice water immersion" and combined with " autotransplantation" to establish EM rat model with SYDBSS. One Milliliter BWHR at 3.33 g/mL was administered to rats in the CM group by gastrogavage. Gestrinone at the daily dose of 0. 5 mg/kg was administered to rats in the WM group by gastrogavage. Equal volume of normal saline was administered to rats in the model group, the blank control group, and the sham-operation group. The size and morphology of ectopic foci in rats were observed after 4 weeks of medication. Expressions of serum CA125, plasma cyclic adenosine monophosphate (cAMP), and plasma cyclic guanosine monophosphate (cGMP) were detected by radioimmunoassay. Morphological changes of eutopic endometrium and ectopic tissue were observed under the optical microscope by HE staining. Protein expressions and contents of HIF-lα, PHD2, and VHL were detected by immunohistochemical SABC method and Western blot. mRNA expressions of HIF-1α, PHD2, and VHL were detected by RT-PCR. RESULTS: The ectopic foci grew significantly in the model group. Their volumes were obviously contracted after treated by CM and WM. Compared with the blank control group and the sham-operation group, serum CA125 and plasma cGMP obviously increased, cAMP obviously decreased (P < 0.05); expressions and contents of HIF-1α mRNA and protein all decreased (P < 0.05); mRNA and protein expressions and contents of PHD2 and VHL all decreased in the model group (P < 0.05). Compared with model group, levels of CA125 and cGMP obviously decreased; cAMP levels obviously increased, expressions and contents of HIF-1α mRNA and protein all increased, mRNA and protein expressions and contents of PHD2 and VHL all increased in the WM group and the CM group (P < 0.05). Compared with the CM group, PHD2 protein contents were higher in the WM group (P < 0.05). HIF-1α was negatively correlated with PHD2 (r = -0.799, P = 0.00). HIF-1α was negatively correlated with VHL (r = -0. 625, P = 0.003). CONCLUSIONS: BWHR could effectively treat EM. Its mechanism might be associated with reducing contents of HIF-1α, serum CA125, and plasma cGMP, and up-regulating expressions of PHD2, VHL, and cAMP.

[Studies on the chemical constituents of Ligularia macrophylla].

OBJECTIVE: To study the chemical constituents of Ligularia macrophylla. METHODS: Isolation and purification were carried out on repeated silica gel column chromatography. The structures of the compounds were identified by physico-chemical properties and spectral analyses. RESULTS: Eight compounds were isolated and identified as kaempferol (1), 2,4'-dihydroxy-5-methoxychalcone (2), 5-hydroxy-3,4', 7-trimethoxyflavone (3), isobutyl ester terephthalic acid (4), 4-hydroxybenzaldehyde (5), mono (2-ethylhexyl) terephthalate (6), lupeol (7), beta-sitosterol (8). CONCLUSION: Compounds 1 - 7 are isolated from this plant for the first time.

[Bioremediation efficiency of applying Daphnia magna and submerged plants: a case study in Dishui Lake of Shanghai, China].

From April 2007 to January 2008, a bioremediation experiment was conducted in a diversion channel of D-port pilot area of Dishui Lake (the channel length is 950 m, and its water volume is 10000 m3). Daphnia magna was first introduced to filter the high biomass of phytoplankton and other particulate organic matter, and then, five submerged plant species Elodea canadensis, Vallisneria spiralis, Hydrilla verticillata, Potamogeton lucens, and Potamogeton crispus were transplanted. Water samples were collected monthly to monitor the water quality and to investigate the bioremediation efficiency. Ten months monitoring data showed that in the remediation area, the water body's total nitrogen (TN), ammonium nitrogen (NH4(+)-N), nitrate nitrogen (NO3(-)-N), nitrite nitrogen (NO2(-)-N), total phosphorus (TP), and reactive phosphate (PO4(3-)-P) concentrations and chemical oxygen demand (COD) were significantly lower (P < 0.01), dissolved oxygen (DO) was increased by 50.4%, and the Secchi depth (SD) reached to an average of 3.4-3.7 m. Overall, the water quality was up to grades II or III of state water quality standards for surface water. In March 2008, the established submerged plant community was used to test its effectiveness in improving the eutrophicated water body from Dishui Lake, and the results showed that after 7-day treatment, except biological oxygen demand (BOD), the TN, TP, NO3(-)-N, NO2(-)-N, NH4(+)-N, and PO4(3-)-P concentrations and COD of the eutrophicated water were all decreased significantly, the DO was increased by 17.98%, and the SD was increased by 30 cm. The present study demonstrated the effectiveness of introducing D. magna and transplanting submerged plants in improving the water quality of Dishui Lake.