Efficacy of acupuncture in refractory irritable bowel syndrome: study protocol for a randomised controlled trial.

INTRODUCTION: Irritable bowel syndrome (IBS) is a common chronic functional gastrointestinal disorder that presents with abdominal pain/discomfort and altered bowel patterns. IBS has multiple potential causes for which conventional medicines have had limited success, resulting in a significant number of patients who do not sensitively respond to pharmacotherapy for a period of 12 months and who develop a continuing symptom profile (described as refractory IBS) and seek help through (non)pharmacological treatments. The aim of this study is to investigate the efficacy and safety of acupuncture therapy for refractory IBS on the basis of conventional treatments. METHODS AND ANALYSIS: A total of 170 eligible patients who meet the Rome IV criteria for refractory IBS will be randomly allocated to receive acupuncture or sham acupuncture. Each patient will receive 12 sessions of acupuncture over 4 weeks and a 4-week follow-up. The primary outcome will be the IBS Symptom Severity Score. Secondary outcomes will include the proportion of participants experiencing adequate relief of global IBS symptoms, the weekly frequency of defecation, the stool properties assessed by the Bristol Grading Scale, the Work and Social Adjustment Scale, the IBS-Quality of Life score, and the Self-Rating Depression Scale and Self-Rating Anxiety Scale anxiety and depression scores. Outcome measures will be collected at baseline, 2 and 4 weeks of the intervention, and 6 and 8 weeks after the intervention. Categorical variables will be compared with Fisher's exact test or the Wilcoxon rank-sum test, and continuous variables will be compared using Student's t-test or the Wilcoxon rank-sum test. ETHICS AND DISSEMINATION: The entire project has been approved by the ethics committees of Beijing University of Chinese Medicine (2020BZYLL0507) and Sichuan Province Regional Institution for Conducting Research on Traditional Chinese Medicine (2020KL-025). The outcomes of the trial will be disseminated through peer-reviewed publications. TRIAL REGISTRATION NUMBER: NCT04276961.

Prognostic Value of an Autophagy-Related Five-Gene Signature for Lower-Grade Glioma Patients.

Background: Molecular characteristics can be good indicators of tumor prognosis and have been introduced into the classification of gliomas. The prognosis of patients with newly classified lower-grade gliomas (LGGs, including grade 2 and grade 3 gliomas) is highly heterogeneous, and new molecular markers are urgently needed. Methods: Autophagy related genes (ATGs) were obtained from Human Autophagy Database (HADb). From the Cancer Genome Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA), gene expression profiles including ATG expression information and patient clinical data were downloaded. Cox regression analysis, receiver operating characteristic (ROC) analysis, Kaplan-Meier analysis, random survival forest algorithm (RSFVH) and stratification analysis were performed. Results: Through univariate Cox regression analysis, we found a total of 127 ATGs associated with the prognosis of LGG patients from TCGA dataset and a total of 131 survival-related ATGs from CGGA dataset. Using TCGA dataset as the training group (n = 524), we constructed a five-ATG signature (including BAG1, BID, MAP1LC3C, NRG3, PTK6), which could divide LGG patients into two risk groups with significantly different overall survival (Log Rank P < 0.001). Then we confirmed in the independent CGGA dataset that the five-ATG signature had the ability to predict prognosis (n = 431, Log Rank P < 0.001). We further discovered that the predictive ability of the five-ATG signature was better than the existing clinical indicators and IDH mutation status. In addition, the five-ATG signature could further classify patients after receiving radiotherapy or chemotherapy into groups with different prognosis. Conclusions: We identified a five-ATG signature that could be a reliable prognostic marker and might be therapeutic targets for autophagy therapy for LGG patients.

Electroacupuncture Improves Cognitive Function in Senescence-Accelerated P8 (SAMP8) Mice via the NLRP3/Caspase-1 Pathway.

Background. Clinically, electroacupuncture (EA) is the most common therapy for aging-related cognitive impairment (CI). However, the underlying pathomechanism remains unidentified. The aims of this study were to observe the effect of EA on cognitive function and explore the potential mechanism by which EA acts on the NLRP3/caspase-1 signaling pathway. Main Methods. Thirty male SAMP8 mice were randomly divided into the model, the 2 Hz EA and 10 Hz EA groups. Ten male SAMR1 mice were assigned to the control group. Cognitive function was assessed through the Morris water maze test. Hippocampal morphology and cell death were observed by HE and TUNEL staining, respectively. The serum IL-1ß, IL-6, IL-18, and TNF-α levels were measured by ELISA. Hippocampal NLRP3, ASC, caspase-1, GSDM-D, IL-1ß, IL-18, Aß, and tau proteins were detected by Western blotting. Key Findings. Cognitive function, hippocampal morphology, and TUNEL-positive cell counts were improved by both EA frequencies. The serum IL-1ß, IL-6, IL-18, and TNF-α levels were decreased by EA treatment. However, 10 Hz EA reduced the number of TUNEL-positive cells in the CA1 region and serum IL-1ß and IL-6 levels more effectively than 2 Hz EA. NLRP3/caspase-1 pathway-related proteins were significantly downregulated by EA, but 2 Hz EA did not effectively reduce ASC protein expression. Interestingly, both EA frequencies failed to reduce the expression of Aß and tau proteins. Significance. The effects of 10 Hz EA at the GV20 and ST36 acupoints on the NLRP3/caspase-1 signaling pathway may be a mechanism by which this treatment relieves aging-related CI in mice.

[Encircling needling is superior to "Bangci"(focal center-side needling) in promoting wound healing in diabetic mice].

OBJECTIVE: To compare the therapeutic effects of "Bangci"(focal center-side needling) and encircling needling in promoting skin wound healing and local blood perfusion in diabetic mice. METHODS: Thirty-two male C57BL/6N mice were randomized into normal, diabetic model, focal center-side needling and encircling needling groups (n=8 in each group). The skin wound model was prepared by cutting a piece of full-thickness skin at the mouse's back by using a puncher. One hour after modeling, two acupuncture needles were respectively inserted into the center of the wound and the spot at the normal skin about 0.5 cm away from the edge of the wound for mice of the focal center-side needling group, followed by EA (0.5 mA, 0.5 Hz) for 30 min. For mice of the encircling needling group, 4 acupuncture needles were respectively inserted into the upper, lower, left and right normal marginal skin around the wound, followed by EA stimulation with the same parameters as those of the center-side needling group. The wound conditions, diameter and area of the wound were monitored, and the wound blood perfusion volume was measured by using a laser speckle flowmeter. RESULTS: The wound shrinkage rates were significantly higher at the time-points of 3, 5, 7, 9 and 11 d after modeling in both focal center-side needling and encircling needling groups than in the model group (P<0.05), and on day 3, 5, 7 and 9 in the encircling needling group than in the focal center-side needling group (P<0.05). The wound healing time was obviously earlier in both focal center-side needling and encircling needling groups than in the model group (P<0.05), and in the encircling needling group than in the focal center-side needling group (P<0.05). Following modeling, the volume of wound blood perfusion was considerably higher from day 1 to 9 and markedly lower on day 11 in the model group than in the normal group (P<0.01), and after the intervention, the blood perfusion volume was considerably decreased on day 3, 5, 7 and 9 in both the focal center-side needling and encircling needling groups(P<0.05, P<0.01), and obviously increased on day 11 in the encircling needling group relevant to the model group (P<0.01).Comparison between post- and pre-EA stimulation showed that the immediate blood perfusion volume was significantly increased from day 1 to 11 after EA stimulation in both the focal center-side needling and encircling needling groups (P<0.01, P<0.05). The therapeutic effect of the encircling needling group was significantly superior to that of the focal center-side needling group in lowering blood perfusion volume from day 3 to 7, and in increasing blood perfusion volume on day 9 and 11 (P<0.01). Under the naked eyes, the conditions of exudation and inflammatory reaction, and the scar and granulation tissue were relatively milder and better respectively in both the center-side needling and encircling needling groups than in the model group. CONCLUSION: Both focal center-side needling and encircling needling can promote the skin wound healing by increasing the blood perfusion in diabetic mice, and the therapeutic effect of the encircling needling method was significantly superior to that of the focal center-side needling method.

Cytoprotective effects of the medicinal herb Astragalus membranaceus on lipopolysaccharide­exposed cells.

Astragalus membranaceus (AM) is a traditional Chinese medicinal herb, whose cytoprotective effects remain largely unknown. Here, the bacterial endotoxin lipopolysaccharide (LPS) was applied to a human pulmonary type II­like epithelial lung adenocarcinoma cell line, a human umbilical vein endothelial cell line, and a human bladder carcinoma cell line to construct in vitro models of intracellular oxidative stress. The authors assayed the cellular and mitochondrial cytoprotective effects of varying doses of AM root extract upon these cell lines. The cell lines were cultured as follows: LPS­only group, four LPS+AM groups treated with various AM concentrations plus LPS, and an untreated control group. Flow cytometry was used to assess apoptosis and cell cycle progression. A 2',7'­dichlorofluorescein­diacetate assay was used to quantitate reactive oxygen species (ROS) generation. Mitochondrial membrane potential (Δψmit) was analyzed by Rhodamine 123 assay. Western blotting was performed to detect cleaved caspase­3, p53, and B cell lymphoma (Bcl)­2 levels. Across all cell lines, LPS significantly elevated apoptosis rates, shifted cells to S/G2 phase, increased ROS production, reduced Δψmit, upregulated cleaved caspase­3, upregulated p53, and downregulated Bcl­2 relative to controls (all P<0.05). As a general trend, increasing AM concentrations produced progressively greater reductions in the apoptosis rate, greater reductions in S/G2 phase %, greater reductions in ROS production, greater increases in Δψmit, greater reductions in cleaved caspase­3 and p53 expression, and greater increases in Bcl­2 expression. AM treatment protects human pulmonary and bladder epithelial cells, in addition to human endothelial cells, from LPS­induced apoptosis, in a dose­dependent manner.