Supplementation with high-GABA-producing Lactobacillus plantarum L5 ameliorates essential tremor triggered by decreased gut bacteria-derived GABA.

BACKGROUND: The γ-aminobutyric acid (GABA) hypothesis posits a role of GABA deficiency in the central nervous system in the pathogenesis and progression of essential tremor (ET). However, the specific causative factor for GABA deficiency is not clear. The gut microbiota in mammals has recently been considered as a significant source of GABA. Furthermore, the GABA-based signals originating from the intestine can be transmitted to the brain through the "enteric nervous system-vagus nerve-brain" axis. However, the plausible contribution of gut microbiota to ET seems inspiring but remains obscure. METHODS: Fecal samples from patients with ET and healthy controls were examined by metagenomic sequencing to compare the composition of gut microbiota and the expression of genes involved in GABA biosynthesis. The impact of gut microbiota on ET was explored through transplantation of fecal microbiota from patients with ET into the murine ET model. Lactic acid bacteria producing high amounts of GABA were identified through whole-genome sequencing and ultra-performance liquid chromatography-tandem mass spectrometry. Subsequently, mice were treated with the high-GABA-producing strain Lactobacillus plantarum L5. Tremor severity, behavioral tests, pro-inflammatory cytokines, GABA concentration, and gut microbiota composition were examined in these mice. RESULTS: The gut microbiota of patients with ET demonstrated an impaired GABA-producing capacity and a reduced fecal GABA concentration. Transplantation of the gut microbiota from patients with ET induced an extension of tremor duration and impaired mobility in the murine model of ET. L5 exhibited an augmented GABA-producing capacity, with the De Man-Rogosa-Sharpe culture broth containing 262 mg/l of GABA. In addition, administration of L5 significantly decreased the tremor severity and enhanced the movement capability and grasping ability of ET mice. In vivo mechanistic experiments indicated that L5 reshaped the gut microbial composition, supplemented the mucosa-associated microbiota with GABA-producing capacity, increased the GABA concentrations in the cerebellum, and diminished inflammation in the central nervous system. CONCLUSIONS: These findings highlight that deficiency of GABA-producing gut microbes plays an essential role in the pathogenesis of ET and that L5 is a promising candidate for treating ET.

[Effect of Danzhi Xiaoyao Powder on behavior and mitochondrial morphology and function of anxiety model rats].

Danzhi Xiaoyao Powder is a classical prescription for anxiety. This study aims to analyze the effect of this medicine on mitochondrial morphology and function of anxiety rats and explore the mechanism of it against anxiety. Specifically, uncertain empty bottle drinking water stimulation(21 days) was employed to induce anxiety in rats. The elevated plus-maze test and open field test were respectively performed on the 7 th, the 14 th, and the 21 st days of the stimulation, so as to detect the anxiety-related protein index brain-derived neurotrophic factor(BDNF) and evaluate the anxiety level of animals. On this basis, the effect of this prescription on anxiety rats was preliminarily evaluated. After the behavioral test on the 21 st day, rats were killed and the brain tissues were separated for the observation of the mitochondrial morphology and the determination of mitochondrial function-related indicators and the adenosine 5'-monophosphate-activated protein kinase(AMPK) level. The results showed that Danzhi Xiaoxiao Powder could alleviate the anxiety-like behavior of rats, significantly increase the percentage of time in open arm in elevated plus-maze test and the ration of activity time in the central area of the field, dose-dependently raise the activity levels of respiratory chain complex Ⅰ,Ⅱ,Ⅲ and Ⅳ and the adenosine triphosphate(ATP) content, and elevate the levels of BDNF and phosphorylated AMPK(p-AMPK). Clear structure and intact morphology of mitochondrial cristae in medial prefrontal cortex cells and amygdala were observed in the Danzhi Xiaoyao Powder group. In summary, Danzhi Xiaoyao Powder exerts therapeutic effect on anxiety, and the mechanism is the likelihood that p-AMPK protects the structure and maintains the function of mitochondria.

Effect of manual soft tissue therapy on the pain in patients with chronic neck pain: A systematic review and meta-analysis.

OBJECTIVE: To systematically evaluate the impact of manual soft tissue therapy (MSTT) on the degree of pain in patients with chronic neck pain (CNP). METHODS: Trials included in the meta-analysis were identified by searching 5 English databases, including the PubMed, Embase, Cochrane Library, Web of Science and U.S. Clinical Trial Registry databases. The search was conducted with the subject terms neck pain, soft tissue treatment, massage, and myofascial release. We assessed the included trials using the Cochrane risk-of-bias tool. STATA statistical software version 16.0 was used for statistical analysis. Additionally, subgroup analysis and sensitivity analysis were performed to analyze the sources of heterogeneity and assess the stability of the research results. Begg's funnel plot and Egger's publication bias plot were used to assess potential publication bias. RESULTS: This systematic review included a total of 12 randomized controlled trials (566 patients in total). The participants were between 18 and 85 years old. Most of the included studies were of medium quality. This meta-analysis validated the effectiveness of MSTT in alleviating pain symptoms in patients with CNP (ES: 0.83; 95% CI: 1.15 to -0.51; P = 0.001). Egger's publication bias plot and Begg's funnel plot indicated that there may be potential publication bias. CONCLUSION: This meta-analysis found that MSTT has a significant effect on alleviating the pain of patients with CNP. In addition, the use of different pain measurement tools may influence effect of the intervention, but more clinical studies are needed in the future to determine the specific effect.

[Effect of manual acupuncture on expression of BDNF/TrkB/ERK/CREB signaling in infarcted tissue of cerebral ischemia rats].

OBJECTIVE: To observe the effect of acupuncture on the expression of cortical brain-derived neurothrophic factor (BDNF) and tyrosine kinase receptor B (TrkB) mRNAs, and phosphorylated extracellular regulated protein kinases (p-ERK) 1/2 and phosphorylated cAMP response element binding protein (p-CREB) proteins in cerebral ischemia (CI) rats, so as to explore its neuroprotective mechanism on ischemic brain tissue. METHODS: Thirty male SD rats were randomly divided into sham-operation (control), model and acupuncture groups, with 10 rats in each group. The CI model was established by occlusion of the left middle cerebral artery (MCAO) using thread embolization method. Manual acupuncture was applied to"Baihui"(GV20),"Quchi"(LI11),"Neiguan"(PC6),"Hegu"(LI4),"Zusanli"(ST36),"Sanyinjiao"(SP6) and"Shenmai"(BL62) once daily, 6 days a week for 2 weeks. The modified neurological severity score (mNSS) including motor, sensation, balance, and neural reflex functions (0 point, normal; 18 points, maximal deficit) was used to assess the neurological impairment state. The expression levels of p-ERK 1/2 and p-CREB proteins, and BDNF and TrkB mRNAs in the ischemic cerebral cortex were measured by Western blot and real-time fluorescence quantitative PCR, respectively. RESULTS: The mNSS was significantly increased on day 1, 7 and 14 in the model group compared with the control group (P<0.01), and evidently decreased on day 14 in the acupuncture group in contrast to the model group (P<0.05). The expression levels of BDNF and TrkB mRNAs, and p-ERK1, p-ERK2 and p-CREB proteins on day 7 and 14 were significantly lower in the model group than in the control group (P<0.001). Following acupuncture treatment, the down-regulated expression levels of the two genes and the three proteins on day 14 were reversed (P<0.001, P<0.05, P<0.01). CONCLUSION: Acupuncture may promote neurological recovery in rats with cerebral ischemia, which may be related to its function in up-regulating the activities of BDNF/TrkB-ERK-CREB signaling in the cerebral cortex on the ischemic side.

Rubus chingii Hu: an overview of botany, traditional uses, phytochemistry, and pharmacology.

Rubus chingii Hu, a member of the rosaceae family, is extensively distributed in China and Japan. Its unripe fruits (Fupenzi in Chinese) have a long history of use as an herbal tonic in traditional Chinese medicine for treating various diseases commonly associated with kidney deficiency, and they are still in use today. Phytochemical investigations on the fruits and leaves of R. chingii indicate the presence of terpenoids, flavonoids, steroids, alkaloids, phenylpropanoids, phenolics, and organic acids. Extracts or active substances from this plant are reported to have various pharmacological properties, including antioxidant, anti-inflammatory, antitumor, antifungal, antithrombotic, antiosteoporotic, hypoglycemic, and central nervous system-regulating effects. This review provides up-to-date information on the botanical characterizations, traditional usages, chemical constituents, pharmacological activities, toxicity, and quality control of R. chingii. Possible directions for future research are also briefly proposed. This review aims to supply fundamental data for the further study of R. chingii and contribute to the development of its clinical use.

Study on the Changes of Chemical Constituents in Different Compatibilities of Ginseng-Prepared Rehmannia Root and Their Effects on Bone Marrow Inhibition after Chemotherapy.

Ginseng (G) and Prepared Rehmannia Root (PRR) are commonly used in traditional Chinese medicine for blood supplementation. This study aimed to study G and PRR with different compatibility ratios changes in chemical composition and inhibition of cyclophosphamide-induced myelosuppression. HPLC was used to determine the chemical constituents of 13 ginsenosides, 5-hydroxymethylfurfural (5-HMF) and verbascoside in different proportions of G-PRR. Balb/c mice were injected intraperitoneally with cyclophosphamide (CTX) to induce bone marrow suppression. The effects of different proportions of G-PRR on peripheral blood, bone marrow nucleated cells, thymus and spleen index of myelosuppressed mice were analyzed. The results showed that the compatibility of G and PRR can promote the dissolution of ginsenosides, and the content of conventional ginsenosides decreased, and the content of rare ginsenosides increased. Different proportions of G-PRR increased the number of peripheral blood and bone marrow nucleated cells in cyclophosphamide-induced bone marrow suppression mice (p < 0.01), increased thymus index (p < 0.01), decreased spleen index (p < 0.01). Different proportions of G-PRR can improve the myelosuppression induced by cyclophosphamide in mice, and the combined effect of G-PRR is better than the single decoction of G and PRR. Among them, G-PRR 2 : 3 and G-PRR 1 : 2 were better than the other groups. These results indicate that different proportion of G-PRR can improve bone marrow suppression, and the combined decoction of G-PRR is better than the separate Decoction in improving bone marrow suppression. This improvement may be related to the changes of the substance basis and active ingredients of G-PRR.

Saikosaponin-d attenuated lipopolysaccharide-induced depressive-like behaviors via inhibiting microglia activation and neuroinflammation.

Saikosaponin-d (SSd), a triterpenoid saponins compound extracted from Radix Bupleuri, has been demonstrated to effectively alleviate chronic mild stress-induced depressive behaviors in rats, but the underlying molecular mechanisms are still uncertain. Increasing evidence indicated that microglia activation and inflammatory responses were involved in the pathogenesis of depression. Thus, we desired to induce inflammation-related depressive-like behaviors in mice by injecting lipopolysaccharide (LPS) to investigate whether the antidepressant effect of SSd is related to inhibiting inflammation. The results of behavioral tests showed that SSd administration ameliorated LPS-induced depressive-like behaviors, as shown by increased sucrose consumption in the sucrose preference test and decreased immobility time in the tail suspension test and forced swimming test. Furthermore, immunostaining results showed that SSd pretreatment inhibited LPS-induced microglia activation in the hippocampus of mice and primary microglia cells. Enzyme-linked immunosorbent assay (ELISA) results showed that SSd pretreatment suppressed LPS-induced overexpression of inflammatory factors such as interleukin (IL)-1ß, IL-6, tumor necrosis factor (TNF)-α both in vivo and in vitro. Immunostaining and western blot analysis results demonstrated that SSd pretreatment also inhibited LPS-induced HMGB1 translocation from nuclear to extracellular and decreased the protein levels of TLR4, p-IκB-α, NF-κBp65. These results suggested that SSd effectively improved LPS-induced inflammation-related depressive-like behaviors by inhibiting LPS-induced microglia activation and neuroinflammation, and the possible mechanism might associate with the regulation of the HMGB1/TLR4/NF-κB signaling pathway.

Global Functional Network Connectivity Disturbances in Parkinson's Disease with Mild Cognitive Impairment by Resting-State Functional MRI.

Examining the spontaneous BOLD activity to understand the neural mechanism of Parkinson's disease (PD) with mild cognitive impairment (MCI) is a focus in resting-state functional MRI (rs-fMRI) studies. This study aimed to investigate the alteration of brain functional connectivity in PD with MCI in a systematical way at two levels: functional connectivity analysis within resting state networks (RSNs) and functional network connectivity (FNC) analysis. Using group independent component analysis (ICA) on rs-fMRI data acquired from 30 participants (14 healthy controls and 16 PD patients with MCI), 16 RSNs were identified, and functional connectivity analysis within the RSNs and FNC analysis were carried out between groups. Compared to controls, patients with PD showed decreased functional connectivity within putamen network, thalamus network, cerebellar network, attention network, and self-referential network, and increased functional connectivity within execution network. Globally disturbed, mostly increased functional connectivity of FNC was observed in PD group, and insular network and execution network were the dominant network with extensively increased functional connectivity with other RSNs. Cerebellar network showed decreased functional connectivity with caudate network, insular network, and self-referential network. In general, decreased functional connectivity within RSNs and globally disturbed, mostly increased functional connectivity of FNC may be characteristics of PD. Increased functional connectivity within execution network may be an early marker of PD. The multi-perspective study based on RSNs may be a valuable means to assess functional changes corresponding to specific RSN, contributing to the understanding of the neural mechanism of PD.

[History of processing of Sojae Semen Praeparatum].

Sojae Semen Praeparatum (SSP) is commonly used as a type of dietetic Chinese herb. By collecting and analyzing ancient and recent literatures, a textual criticism was conducted on the historical evolution of the processing of SSP. Fermented soybean was recorded in Shijing, and relevant rational processing was described in Qimin Yaoshu. In the early time, fermented soybean included the type of "salty" and "light". After the Ming Dynasty, "light" fermented soybean or SSP was recognized as a better medicinal matter than salty fermented soybean, and the fermentation processing was recorded more clearly. In modern time, many characteristic methods for processing SSP have been developed. Today, the processing of SSP is mainly based on the Chinese Pharmacopoeia, which records soybean as a main ingredient and Artemisiae Annuae Herba, Mori Folium as excipients.

Salvia miltiorrhiza Bunge (Danshen) extract attenuates permanent cerebral ischemia through inhibiting platelet activation in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Danshen is a crude herbal drug isolated from dried roots of Salvia miltiorrhiza Bunge. This plant is widely used in oriental medicine for the treatment of cardiovascular and cerebrovascular diseases. The supercritical CO2 extract from Danshen (SCED) (57.85%, 5.67% and 4.55% for tanshinone IIA, tanshinone I and cryptotanshinone respectively) was studied in this article, whose potential molecular mechanism remains unclear, especially in anti-thrombosis. AIM OF THE STUDY: The present study was designed to observe the protective effect of SCED on ischemic stroke in rats and to explore the underlying anti-thrombosis mechanism. MATERIALS AND METHODS: Following induction of cerebral ischemia in rats by permanent middle cerebral artery occlusion (pMCAO). Neurological defect score, cerebral blood flow, infarct size, and brain edema were measured to evaluate the injury. Arteriovenous shunt thrombosis model and adenosine 5'-diphosphate (ADP) induced acute pulmonary embolism model were conducted to estimate the antithrombotic effect of SCED. In order to investigate the effects of SCED on platelet aggregation, rat platelet-rich-plasma (PRP) were incubated with SCED prior to the addition of the stimuli (ADP or 9, 11-dideoxy-11α, 9α-epoxymethanoprostaglandin F2α (U46619)). Aggregation was monitored in a light transmission aggregometer. Inhibitory effect of SCED on thromboxane A2 (TXA2) release was detected by ELISA kit. Phospholipase C (PLC)/ Protein kinase C (PKC) signaling pathway was analyzed by a Western blot technique. The effect of the SCED was also studied in vivo on bleeding time in mice. RESULTS: SCED improved the neurological defect score, increased cerebral blood flow, reduced infarct size and alleviated brain edema in rats exposed to pMCAO. After administration of SCED, thrombosis formation in arteriovenous shunt was inhibited and recovery time in pulmonary embolism was shortened. The inhibitory effect of SCED on platelet activation was further confirmed by TXB2 ELISA kit and Western blot analysis of PLC/PKC signaling pathway. CONCLUSIONS: SCED attenuates cerebral ischemic injury. The possible mechanism is that SCED inhibits thrombosis formation, platelet aggregation and activation of PLC/PKC pathway. On this basis, this new extract could be a promising agent to inhibit thrombosis formation and protect against cerebral ischemia injury.

Fabrication and characterization of antioxidant pickering emulsions stabilized by zein/chitosan complex particles (ZCPs).

Lipid peroxidation in oil-in-water (o/w) emulsions leads to rancidity and carcinogen formation. This work attempted to protect lipid droplets of emulsions from peroxidation via manipulation of the emulsions' interface framework using dual-function zein/CH complex particles (ZCPs). ZCP with intermediate wettability was fabricated via a simple antisolvent approach. Pickering emulsions were produced via a simple and inexpensive shear-induced emulsification technique. ZCP was irreversibly anchored at the oil-water interface to form particle-based network architecture therein, producing ultrastable o/w Pickering emulsions (ZCPEs). ZCPE was not labile to lipid oxidation, evidenced by low lipid hydroperoxides and malondialdehyde levels in the emulsions after thermally accelerated storage. The targeted accumulation of curcumin, a model antioxidant, at the interface was achieved using the ZCP as interfacial vehicle, forming antioxidant shells around dispersed droplets. The oxidative stability of ZCPEs was further improved. Interestingly, no detectable hexanal peak appeared in headspace gas chromatography of the Pickering emulsions. The novel interfacial architecture via the combination of steric hindrance from ZCP-based membrane and interfacial cargo of curcumin endowed the emulsions with favorable oxidative stability. This study opens a promising pathway for producing antioxidant emulsions via the combination of Pickering stabilization mechanism and interfacial delivery of antioxidant.

GAMETOPHYTE DEFECTIVE 1, a putative subunit of RNases P/MRP, is essential for female gametogenesis and male competence in Arabidopsis.

RNA biogenesis, including biosynthesis and maturation of rRNA, tRNA and mRNA, is a fundamental process that is critical for cell growth, division and differentiation. Previous studies showed that mutations in components involved in RNA biogenesis resulted in abnormalities in gametophyte and leaf development in Arabidopsis. In eukaryotes, RNases P/MRP (RNase mitochondrial RNA processing) are important ribonucleases that are responsible for processing of tRNA, and transcription of small non-coding RNAs. Here we report that Gametophyte Defective 1 (GAF1), a gene encoding a predicted protein subunit of RNases P/MRP, AtRPP30, plays a role in female gametophyte development and male competence. Embryo sacs were arrested at stages ranging from FG1 to FG7 in gaf1 mutant, suggesting that the progression of the gametophytic division during female gametogenesis was impaired in gaf1 mutant. In contrast, pollen development was not affected in gaf1. However, the fitness of the mutant pollen tube was weaker than that of the wild-type, leading to reduced transmission through the male gametes. GAF1 is featured as a typical RPP30 domain protein and interacts physically with AtPOP5, a homologue of RNases P/MRP subunit POP5 of yeast. Together, our data suggest that components of the RNases P/MRP family, such as RPP30, play important roles in gametophyte development and function in plants.