Rapid Mycobacterium abscessus antimicrobial susceptibility testing based on antibiotic treatment response mapping via Raman Microspectroscopy.

OBJECTIVES: Antimicrobial susceptibility tests (ASTs) are pivotal tools for detecting and combating infections caused by multidrug-resistant rapidly growing mycobacteria (RGM) but are time-consuming and labor-intensive. DESIGN: We used a Mycobacterium abscessus-based RGM model to develop a rapid (24-h) AST from the beginning of the strain culture, the Clinical Antimicrobials Susceptibility Test Ramanometry for RGM (CAST-R-RGM). The ASTs obtained for 21 clarithromycin (CLA)-treated and 18 linezolid (LZD)-treated RGM isolates. RESULTS: CAST-R-RGM employs D2O-probed Raman microspectroscopy to monitor RGM metabolic activity, while also revealing bacterial antimicrobial drug resistance mechanisms. The results of clarithromycin (CLA)-treated and linezolid (LZD)-treated RGM isolates exhibited 90% and 83% categorical agreement, respectively, with conventional AST results of the same isolates. Furthermore, comparisons of time- and concentration-dependent Raman results between CLA- and LZD-treated RGM strains revealed distinct metabolic profiles after 48-h and 72-h drug treatments, despite similar profiles obtained for both drugs after 24-h treatments. CONCLUSIONS: Ultimately, the rapid, accurate, and low-cost CAST-R-RGM assay offers advantages over conventional culture-based ASTs that warrant its use as a tool for improving patient treatment outcomes and revealing bacterial drug resistance mechanisms.

"Huoling Shengji granule" for amyotrophic lateral sclerosis: protocol for a multicenter, randomized, double-blind, riluzole parallel controlled clinical trial.

Background: There is still a large demand for effective treatments to delay disease deterioration in amyotrophic lateral sclerosis (ALS). Typical symptoms of ALS are considered "flaccidity syndrome" in traditional Chinese medicine (TCM). Huoling Shengji Granule (HLSJ) is a TCM formula used to treat flaccidity syndrome. Results of preclinical tests and a previous clinical study support HLSJ as a novel drug for ALS patients. This trial proposed to examine whether a 48-week course of HLSJ is effective and safe for ALS patients diagnosed with the Chinese medicine syndrome of spleen qi insufficiency and kidney yang deficiency. Methods and analysis: In this phase II, multicenter, randomized, double-blind, riluzole parallel-controlled, superiority-design study, eligible participants had the equal opportunity to be assigned to receive either HLSJ or riluzole randomly. Eleven specialized ALS centers in Mainland China will recruit 144 patients for this trial. The primary and secondary outcomes included the change in the ALSFRS-R score and the Rasch-Built Overall Amyotrophic Lateral Sclerosis Disability Scale (ROADS) from baseline to Week 48. Discussion: Here, we endeavored to evaluate TCM for ALS using a standard evidence-based approach for the first time. In addition, the ROADS, a self-report linear-weighted questionnaire, was selected as a secondary outcome measure. We expect to offer a new reference for the outcome evaluation of ALS trials.Clinical trial registration:http://www.Chictr.org.cn, identifier ChiCTR2100044085.

Development of fermented sea buckthorn (Hippophae rhamnoides L.) juice and investigation of its antioxidant and antimicrobial activity.

Sea buckthorn (Hippophae rhamnoides L.) is an edible and medicinal plant species. However, due to its sour taste, it is not readily accepted by consumers. To overcome this, fermentation can be used to change its flavor profile. In this study, we used response surface methodology (RSM) to determine the best process for producing fermented sea buckthorn juice (FSBJ) using probiotics. The biological enzyme activity and total flavonoid content (TFC) of sea buckthorn juice (SBJ) increased after fermentation. When the number of bacteria inoculated was 4.08 × 106 CFU/mL and the inoculation ratio was 30% Z. mobilis, 5% L. casei, 13.75% L. plantarum, 31.25% P. acidilactici, 12.5% L. animalis, and 7.5% P. pentosaceus, the amount of sugar was 2.98% (w/v) after 20 h of fermentation at 37°C, and the superoxide dismutase (SOD) activity reached 725.44 U/mL, and the TFC reached 2.38 mg/mL. FSBJ demonstrated strong antimicrobial activity against Escherichia coli, Staphylococcus aureus and Botrytis cinerea. Then, to investigate the antioxidant capacity of FSBJ, we used H2O2 to induce oxidative stress in C2C12 cells and assessed the protection conferred by FSBJ to damaged cells. It was discovered that after 24 h of treatment with FSBJ, not only was there an increase in the activities of intracellular SOD and glutathione peroxidase (GSH-Px), but also a reduction in reactive oxygen species (ROS) content, catalase (CAT) activity, and malondialdehyde (MDA) content. This research lays the theoretical groundwork and provides reference materials for the improved fermentation of sea buckthorn and demonstrates its resulting antioxidant effect.

STUDY ON THE INFLUENCE OF DRY DENSITY AND PARTICLE SIZE FRACTAL DISTRIBUTION ON THE RADON CONTROL PERFORMANCE OF OVERLYING SOIL IN URANIUM TAILINGS POND.

In order to explore the influence of dry density and particle size fractal distribution on the radon control performance of overlying soil in uranium tailings pond, overlying soil samples with different particle size fractal dimension and dry density were prepared for radon exhalation experiments. According to the principle of radon generation and diffusion, a set of radon measuring device was designed independently. In addition, the radon concentration on the surface of different overlying soil layers was measured by local static method, and the radon exhalation rate was further calculated. The relationship between radon exhalation law, dry density and fractal dimension of overlying soil was studied by correlation analysis method. Furthermore, a piecewise linear function between radon exhalation rate, dry density and fractal dimension was constructed. Then, the average value of the experimental data was compared with the calculated results. Results show that: at the same dry density and different fractal dimension, the radon exhalation rate decreased with the increase of fractal dimension. When the fractal dimension increased to a certain value, the radon exhalation rate tended to be stable. At the same fractal dimension and different dry density, the radon exhalation rate decreased with the increase of dry density. The calculated results of piecewise linear function were relatively close to the experimental data, which verified the practicability of the formula.

An Integrated Analysis of Network Pharmacology and Experimental Validation to Reveal the Mechanism of Chinese Medicine Formula Naotaifang in Treating Cerebral Ischemia-Reperfusion Injury.

BACKGROUND: Cerebral ischemia-reperfusion injury (CIRI) is a crucial factor leading to a poor prognosis for ischemic stroke patients. As a novel Chinese medicine formula, Naotaifang (NTF) was proven to exhibit a neuroprotective effect against ischemic stroke, clinically, and to alleviate CIRI in animals. However, the mechanisms underlying the beneficial effect have not been fully elucidated. METHODS: In this study, we combined a network pharmacology approach and an in vivo experiment to explore the specific effects and underlying mechanisms of NTF in the treatment of ischemia-reperfusion injury. A research strategy based on network pharmacology, combining target prediction, network construction, gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and molecular docking was used to predict the targets of NTF in treating the ischemic stroke and CIRI. On the other hand, we used HPLC and HRMS to identify biologically active components of NTF. Middle cerebral artery occlusion models in rats were utilized to evaluate the effect and the underlying mechanisms of NTF against CIRI after ischemic stroke. RESULTS: Network pharmacology analysis revealed 43 potential targets and 14 signaling pathways for the treatment of NTF against CIRI after ischemic stroke. Functional enrichment analysis showed that a STAT3/PI3K/AKT signaling pathway serves as the target for in vivo experimental study validation. The results of animal experiments showed that NTF significantly alleviated CIRI by decreasing neurological score, infarct volume, numbers of apoptotic neuronal cells, increasing density of dendritic spines and survival of neurons. Furthermore, NTF could increase the expression of p-STAT3, PI3K, p-AKT. In addition, the detection of apoptosis-related factors showed that the NTF could raise the expression of Bcl-2 and reduce the expression of Bax. CONCLUSION: This network pharmacological and experimental study indicated that NTF, as a therapeutic candidate for the management of CIRI following ischemic stroke, may exert a protective effect through the STAT3/PI3K/AKT signaling pathway.

Effects of the Cistanche tubulosa Aqueous Extract on the Gut Microbiota of Mice with Intestinal Disorders.

Disorders of the gut microbiota are associated with many diseases. The aqueous extract from Cistanche tubulosa (CT), a traditional Chinese herbal formula, has been reported to play a role in protecting the human intestine. However, little is known about its effects on the gut microbiota. The present study was carried out to determine whether the CT aqueous extract can modulate the gut microbiome in mice with intestinal disorders. We found that the damaged intestinal morphology resulting from treatment with cefixime could be rescued using the CT aqueous extract. The comparison of microbial diversity between mice treated with the CT extract and control mice also indicated that the disorder in the microbiome community of model groups could be restored by treatment with high and medium concentrations of the CT aqueous extract. Treatment with cefixime led to a significant decrease in lactic acid bacteria; however, the supplementation of the CT aqueous extract recovered the growth of these lactic acid bacteria. Furthermore, the CT aqueous extract was able to moderate the dramatic changes in the metabolic pathways of the gut microbiome induced by cefixime. These findings provided an insight into the beneficial effects of the CT aqueous extract on gut microbiota, and they also provided an important reference for the development of related drugs in the future.

In Silico and In Vivo Studies on the Mechanisms of Chinese Medicine Formula (Gegen Qinlian Decoction) in the Treatment of Ulcerative Colitis.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease, and Gegen Qinlian Decoction (GQD), a Chinese botanical formula, has exhibited beneficial efficacy against UC. However, the mechanisms underlying the effect of GQD still remain to be elucidated. In this study, network pharmacology approach and molecular docking in silico were applied to uncover the potential multicomponent synergetic effect and molecular mechanisms. The targets of ingredients in GQD were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Bioinformatics Analysis Tool for Molecular mechANism of TCM (BATMAN-TCM) database, while the UC targets were retrieved from Genecards, therapeutic target database (TTD) and Online Mendelian Inheritance in Man (OMIM) database. The topological parameters of Protein-Protein Interaction (PPI) data were used to screen the hub targets in the network. The possible mechanisms were investigated with gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Molecular docking was used to verify the binding affinity between the active compounds and hub targets. Network pharmacology analysis successfully identified 77 candidate compounds and 56 potential targets. The targets were further mapped to 20 related pathways to construct a compound-target-pathway network and an integrated network of GQD treating UC. Among these pathways, PI3K-AKT, HIF-1, VEGF, Ras, and TNF signaling pathways may exert important effects in the treatment of UC via inflammation suppression and anti-carcinogenesis. In the animal experiment, treatment with GQD and sulfasalazine (SASP) both ameliorated inflammation in UC. The proinflammatory cytokines (TNF-α, IL-1ß, and IL-6) induced by UC were significantly decreased by GQD and SASP. Moreover, the protein expression of EGFR, PI3K, and phosphorylation of AKT were reduced after GQD and SASP treatment, and there was no significance between the GQD group and SASP group. Our study systematically dissected the molecular mechanisms of GQD on the treatment of UC using network pharmacology, as well as uncovered the therapeutic effects of GQD against UC through ameliorating inflammation via downregulating EGFR/PI3K/AKT signaling pathway and the pro-inflammatory cytokines such as TNF-α, IL-1ß and IL-6.

Tetrandrine inhibits proliferation of colon cancer cells by BMP9/ PTEN/ PI3K/AKT signaling.

Despite advances in screening and treatment, colon cancer remains one of the leading causes of cancer-related death. Finding novel and useful drug treatment targets is also an urgent need for clinical applications. Tetrandrine (Tet) is extracted from the Chinese medicinal herbal medicine, which is a well-known calcium blocker with a variety of pharmacological activities, including anti-cancer. In this study, we recruited cell viability assay, flow cytometry analysis, cloning formation to confirm that Tet can inhibit the proliferation of SW620 cells, and induce apoptosis. Mechanically, we confirmed that Tet up-regulates the mRNA and protein level of BMP9 in SW620 cells. Over-expression BMP9 enhances the anti-cancer effects of Tet in SW620 cells, but these effects can be partly reversed by silencing BMP9. Also, Tet reduces phosphorylation of Aktl/2/3 in SW620 cells, which could be elevated by overexpressed BMP9 and impaired by silencing BMP9. Furthermore, we demonstrated that Tet reduces phosphorylated PTEN, which can be promoted by overexpressed BMP9, analogously also be attenuated through silencing BMP9. Finally, we introduced a xenograft tumor model to investigate the anti-proliferative effect of Tet, further to explore the effects of BMP9 and PTEN in SW620 cells. Our findings suggested that the anti-cancer activity of Tet in SW620 cells may be mediated partly by up-regulating BMP9, followed by inactivation PI3K/Akt through up-regulating PTEN at least.

Experimental study on permeability characteristics and radon exhalation law of overburden soil in uranium tailings pond.

The permeability characteristics of overburden soil have a significant effect on radon exhalation in uranium tailings pond. To understand this change, the effects of dry density, particle size range, and moisture content on the gas permeability and radon exhalation were studied by correlation analysis method. According to the mechanism of radon generation, the permeability characteristics of overburden, and the weather conditions, a theoretical model of radon exhalation was constructed and an experimental setup was designed. A series of tests on different properties of overburden soil were conducted by uranium tailings permeameter, and the radon exhalation rate of corresponding overburden surface was measured by local static method. Results show that the permeability of overburden decreased, which led to the decrease of radon exhalation: (1) At the same moisture content, with the increase of dry density, both the gas permeability and radon exhalation rate decrease. (2) At the same moisture content, with the increase of the particle size range, both the gas permeability and radon exhalation rate increase. (3) At the same dry density, with the increase of moisture content, the gas permeability decreases, while the radon exhalation rate increases slightly at first and then decreases.

Electroacupuncture ameliorates cerebral ischemia/reperfusion injury by suppressing autophagy via the SIRT1-FOXO1 signaling pathway.

Cerebral ischemia/reperfusion (CIR) injury occurs when blood flow is restored in the brain, causing secondary damage to the ischemic tissues. Previous studies have shown that electroacupuncture (EA) treatment contributes to brain protection against CIR injury through modulating autophagy. Studies indicated that SIRT1-FOXO1 plays a crucial role in regulating autophagy. Here we investigated the mechanisms underlying the neuroprotective effect of EA and its role in modulating autophagy via the SIRT1-FOXO1 signaling pathway in rats with CIR injury. EA pretreatment at "Baihui", "Quchi" and "Zusanli" acupoints (2/15Hz, 1mA, 30 min/day) was performed for 5 days before the rats were subjected to middle cerebral artery occlusion, and the results indicated that EA pretreatment substantially reduced the Longa score and infarct volume, increased the dendritic spine density and lessened autophagosomes in the peri-ischemic cortex of rats. Additionally, EA pretreatment also reduced the ratio of LC3-II/LC3-I, the levels of Ac-FOXO1 and Atg7, and the interaction of Ac-FOXO1 and Atg7, but increased the levels of p62, SIRT1, and FOXO1. The above effects were abrogated by the SIRT1 inhibitor EX527. Thus, we presume that EA pretreatment elicits a neuroprotective effect against CIR injury, potentially by suppressing autophagy via activating the SIRT1-FOXO1 signaling pathway.

Natural history and clinical features of sporadic amyotrophic lateral sclerosis in China.

OBJECTIVES: To describe the natural history and clinical features of sporadic amyotrophic lateral sclerosis (ALS) in Chinese patients, and to report data on the prognostic factors for survival. METHODS: All patients referred to our ALS centre between 2003 and 2012 were followed up every 3 months. Survival and tracheotomy were predefined as primary outcome measures. Group differences were analysed using parametric and non-parametric tests as appropriate. Survival was analysed using the Kaplan-Meier method and Cox regression analysis. RESULTS: Of the 1624 patients with ALS, 75.1% had limb-onset, 14.0% had bulbar-onset, 7.8% had flail-arm syndrome (FAS), 2.6% had progressive muscular atrophy and 0.5% had primary lateral sclerosis. The male:female ratio was 1.7:1, and the mean age at onset was 49.8 years. The median diagnostic delay was 14 months, and the median survival time after symptom onset was 71 months. Male gender, older age at symptom onset, lower body mass index, shorter diagnostic delay, bulbar-onset ALS phenotype, higher Airlie House category at presentation, rural place of residence, use of traditional Chinese medicine and a history of contact with pesticides were associated with poorer survival, whereas female gender or an FAS phenotype may have a better prognosis. CONCLUSIONS: The clinical characteristics and outcomes of Chinese patients with sporadic ALS were different compared with patients from other countries. Compared with other studies, the age at onset of Chinese patients was earlier, the percentage of bulbar-onset ALS was lower and the prognosis was better. This study substantially advances the understanding of the clinical features and epidemiology of this rare disease.

[Advances in the pathway and molecular mechanism for the biodegradation of microcystins].

With increasing discharge of wastewater containing nitrogen and phosphorus into rivers and lakes, harmful cyanobacterial blooms have become more frequent worldwide. The main harm of cyanobacterial blooms is producing and releasing a great amount of algal toxins mainly containing microcystins (MCs). Since MCs are extremely harmful to plants and animals and difficult to be removed efficiently by the traditional processing methods, how to control harmful cyanobacterial blooms and remove MCs have become an unsolved problem in the field of environmental science all over the world. This paper summarized the structure and toxicity of MCs, the MCs-biodegrading bacterial strains, the enzymes, the genes, and the biodegradation pathway and molecular mechanism of MCs. The further research subjects were also proposed. It was hoped that this review could provide a reference for restoring MCs-polluted lakes and reservoirs and ensuring drinking water safety.

Transcriptome analysis of grain-filling caryopses reveals involvement of multiple regulatory pathways in chalky grain formation in rice.

BACKGROUND: Grain endosperm chalkiness of rice is a varietal characteristic that negatively affects not only the appearance and milling properties but also the cooking texture and palatability of cooked rice. However, grain chalkiness is a complex quantitative genetic trait and the molecular mechanisms underlying its formation are poorly understood. RESULTS: A near-isogenic line CSSL50-1 with high chalkiness was compared with its normal parental line Asominori for grain endosperm chalkiness. Physico-biochemical analyses of ripened grains showed that, compared with Asominori, CSSL50-1 contains higher levels of amylose and 8 DP (degree of polymerization) short-chain amylopectin, but lower medium length 12 DP amylopectin. Transcriptome analysis of 15 DAF (day after flowering) caryopses of the isogenic lines identified 623 differential expressed genes (P < 0.01), among which 324 genes are up-regulated and 299 down-regulated. These genes were classified into 18 major categories, with 65.3% of them belong to six major functional groups: signal transduction, cell rescue/defense, transcription, protein degradation, carbohydrate metabolism and redox homeostasis. Detailed pathway dissection demonstrated that genes involved in sucrose and starch synthesis are up-regulated, whereas those involved in non-starch polysaccharides are down regulated. Several genes involved in oxidoreductive homeostasis were found to have higher expression levels in CSSL50-1 as well, suggesting potential roles of ROS in grain chalkiness formation. CONCLUSION: Extensive gene expression changes were detected during rice grain chalkiness formation. Over half of these differentially expressed genes are implicated in several important categories of genes, including signal transduction, transcription, carbohydrate metabolism and redox homeostasis, suggesting that chalkiness formation involves multiple metabolic and regulatory pathways.