Factors influencing the quality of acupuncture clinical trials: a qualitative interview of stakeholders.

OBJECTIVE: To investigate the influencing factors on the quality of acupuncture clinical trials from the stakeholders, and to provide references for improving the quality of acupuncture clinical trials. METHODS: A qualitative study based on semi-structured interviews was performed. Experts, acupuncturists, editors, and patients were interviewed. The interview results were thematically analyzed from transcribed audio recordings. RESULTS: A total of 38 stakeholders were interviewed, including 12 experts, 14 acupuncturists, 2 editors, and 10 patients. There were 25 tree nodes and 106 sub-nodes, with 1141 reference points. The key factors influencing the quality of acupuncture clinical trials could be divided into five core theme frameworks: a) trial design, b) trial conduction, c) research results reporting and publication, d) research evidence dissemination, and e) research evidence transformation and application. CONCLUSIONS: The results reveal that to improve the quality of acupuncture trials, it should consider each step of trial design, trial conduction, research results reporting and publication, research evidence dissemination, and research evidence transformation and application. A guideline for quality control of the whole process of acupuncture clinical trials is needed.

Polysaccharides from Fu brick tea ameliorate obesity by modulating gut microbiota and gut microbiota-related short chain fatty acid and amino acid metabolism.

Fu brick tea (FBT) is a traditional tea manufactured by solid-state fermentation of tea leaves (Camellia sinensis). Although anti-obesity effects have been reported for FBT, the associated role of FBT polysaccharides (PSs) and the underlying mechanisms remain unknown. In this study, we found that FBTPSs inhibited obesity, hyperlipidemia, and inflammation; improved intestinal barrier function; and alleviated gut microbiota dysbiosis in high-fat diet-fed rats. Akkermansia muciniphila, Bacteroides, Parasutterella, Desulfovibrio, and Blautia were the core microbes regulated by FBTPSs. FBTPSs regulated the production of gut microbiota-related metabolites, including short-chain fatty acids (SCFAs), branched-chain amino acids, and aromatic amino acids throughout the development of obesity, and regulated the SCFA-GPR signaling pathway. FBTPS-treated fecal microbiota transplant ameliorated obesity, alleviated gut microbiota dysbiosis, and improved gut microbiota-associated metabolites, suggesting that the anti-obesity effect of FBTPSs was gut microbiota-dependent. FBTPSs may serve as novel prebiotic agents for the treatment of obesity and dysbiosis of gut microbiota.

Observational studies of traditional Chinese medicine may provide evidence nearly consistent with the randomized controlled trials: A meta-epidemiological study.

Background: In traditional Chinese medicine (TCM) field, the benefits of observational studies was more significant. Whether the evidence from observational studies agreed with RCTs in the field of TCM was still unclear. Methods: A meta-epidemiological study was conducted. Meta-analyses and systematic reviews including cohort studies and case-control studies of TCM were included. Ratio of odds ratio (ROR) of randomized controlled trials and observational studies were calculated individually and intercomparisons were conducted by pool analysis. Results: A total of 11 studies and 30 outcome pairs were included in the pool analysis. Using results from the observational studies as the reference group, the polled ROR comparing randomized controlled trials with observational studies was 1.23 (95% confidence interval 1.05 to 1.44, and 95% prediction interval 0.90 to 1.68). The ROR by subgroup analysis were 1.15 (95% confidence interval 0.96 to 1.38; 95% prediction interval 0.95 to 1.39) and 1.12 (95% confidence interval 0.86 to 1.46; 95% prediction interval 0.51 to 2.47) for cohort studies and case-control studies, respectively. Conclusions: There is difference in pooled results between randomized controlled studies and observational studies on TCM. However, the prediction interval shows the difference is small, which suggests observational studies of TCM can be included in data analysis to provide evidence for TCM. Future studies are needed to verify the above conclusion.

Clinical Application of Traditional Chinese Medicine Therapy for Type 2 Diabetes Mellitus: An Evidence Map.

Type 2 diabetes mellitus (T2DM), a common disease with a complex etiology in the world, is an important risk factor for severe cardiovascular and cerebrovascular diseases. However, treatments of T2DM are mainly based on Western medicine, whose severe side effects make traditional Chinese medicine (TCM) therapy more appealing to patients and clinicians. The overall clinical evidence for different TCM therapies in the treatment of T2DM is still unclear. This study aimed to adopt the evidence-mapping method and integrate the evidence from various researches on this topic, to depict the whole picture of TCM therapies for T2DM. This review included searches of PubMed, Embase, Web of Science, and three major Chinese literature databases (CNKI, VIP, and Wanfang) from inception to November 18, 2021. Two independent reviewers screened the literature, extracted information, and evaluated the quality of all included studies. A systematic review was subsequently performed. In total, 47 studies were reviewed, of which 46 studies (97.9%) were from China and 1 (2.1%) was from Canada. The evidence map was conducted according to different TCM therapies, including herbs or herbal extracts, compounds, powders, decoctions, pills, external treatment, basic theories and treatment principles of TCM, proprietary Chinese medicines, and unspecified TCM integrated therapies. According to the AMSTAR-2 scoring results, 4 papers were rated as high quality, 11 were low quality, and 32 were very low quality. Outcome indicators mainly focused on FBG, HbA1c, 2-h PBG, TC, TG, LDL-C, etc. The results showed that different types of TCM treatment had different improvement effects on the outcome indicators of T2DM. More consistent benefits were observed in the improvement of FBG, HbA1c, and 2-h PBG with treatment regimens based on basic theories and treatment principles of TCM, decoctions and pills, and unspecified TCM integrated therapies. Among herbs, ginger and Coptis root showed more improvement in all outcomes. Compounds, powders, and external treatment showed relatively consistent beneficial effects on the improvement of FBG. No serious adverse events were reported. Overall, the current evidence map provided an intuitive overview of the beneficial effects of TCM therapies in the treatment of type 2 diabetes. This study can be used as a reference for the clinical application of traditional Chinese medicine in T2DM, but due to the low-quality level of the included studies, it should be treated with caution in clinical practices.

Alteration of intestinal microecology by oral antibiotics promotes oral squamous cell carcinoma development.

Oral antibiotics can influence cancers and immunotherapy by interfering with the intestinal microbiota. However, the association between oral antibiotics and oral squamous cell carcinoma (OSCC) as well as the mechanisms underlying the effects of oral antibiotics on OSCC remain unclear. Here, we found that oral antibiotics cocktail (4Abx) promoted the tumor development and shifted the microbiota, decreasing the abundance of probiotic bacteria, and altered microbial metabolism in the gut of OSCC mice, increasing tyrosine and decreasing glutamate levels. In vitro experiments showed that tyrosine upregulated the PD-1 expression in T cells, SCC7 cell proliferation, and necroptosis expression. IL-10 expression level in CD11c+ cells was reduced by glutamate. Furthermore, the expression of the necroptosis-related proteins, including receptor-interacting protein kinase 1 (RIPK1), RIPK3, and mixed lineage kinase domain-like (MLKL), was higher in the OSCC mice treated with 4Abx. Supplementation with glutamate or healthy mouse feces by gavage alleviated the tumor-promoting effect of 4Abx with restored balance of microbial metabolism. Overall, we identified the detrimental role of oral antibiotics in promoting OSCC development through altered intestinal microbiota, microbial metabolism, and immune dysbiosis, implying the need for antibiotic stewardship in OSCC treatment.

Synthesis, Cytotoxicity, Anti-Migration and Anti-Invasion Activity of Diphyllin Heterocyclic Derivatives.

BACKGROUND: Diphyllin, an arylnaphthalene lignan lactone, isolated from many traditional medicinal plants, has been reported to possess anticancer and antiviral activities. Natural diphyllin and its glycosides were identified as potent vacuolar H+-ATPase (V-ATPase) inhibitors. OBJECTIVE: The aim of this study was to design and synthesize a series of heterocyclic derivatives of diphyllin as novel anticancer agents. METHODS: The targeted heterocyclic derivatives of diphyllin were synthesized from diphyllin employing etherification reaction and N-substitution reaction. Cytotoxicity of these compounds on four cancer cells was assessed by MTT assay. The inhibitory activity of V-ATPase of compound 3n was measured on MGC-803 cells. Anti-migration and anti-invasion abilities were assessed by transwell invasion assay and scratch wound assay. RESULTS: Most of these derivatives displayed potent cytotoxicity on four cancer cells at submicromolar concentrations. The most potent derivative 3n has been shown to inhibit V-ATPase activity, migration and invasion abilities on MGC-803 cells at 0.75 µM. CONCLUSION: The collective results clearly indicate that heterocyclic derivatives of diphyllin inhibit the viability, V-ATPase activity, migration and invasion of the MGC803 cells. The current findings provide valuable insights for the future development of novel diphyllin derivatives as anticancer agents.

Arginine accelerates intestinal health through cytokines and intestinal microbiota.

Arginine supplementation improves intestinal damage and intestinal immunity, but the underlying mechanism of the effects of arginine supplementation on intestinal SIgA secretion is largely unknown. Therefore, this study was conducted to investigate the underlying pathway on the effects of arginine supplementation in secretory IgA (SIgA) production in mice. The results showed that 0.4% arginine supplementation promoted (P < 0.05) SIgA production in intestinal lumina and IgA+ plasma cell numbers in the ileum of mouse. Arginine supplementation significantly increased (P < 0.05) cytokines expression in mouse ileal associated with T cell-dependent and T cell-independent pathways of SIgA induction, including IL-5, IL-6, IL-13, transforming growth factor (TGF-)ß2, TGF-ß3, TGF-ßR2, a proliferation-inducing ligand (APRIL), B cell-activating factor (BAFF), vasoactive intestinal peptide (VIP) receptor, and retinal dehydrogenases. Further study showed that 0.4% arginine supplementation markly decreased (P < 0.05) bacterial loads in mouse mesenteric lymph nodes and increased bacterial invasion into the mouse ileal wall, while supplementation of antibiotic abrogated the influence of arginine supplementation on SIgA secretion. Therefore, these data suggest that arginine supplementation might promote SIgA secretion through cytokines and intestinal microbiota might play an important role in SIgA secretion by arginine supplementation in the mouse intestine.

N-Acetyl-L-cysteine Protects the Enterocyte against Oxidative Damage by Modulation of Mitochondrial Function.

The neonatal small intestine is susceptible to damage caused by oxidative stress. This study aimed to evaluate the protective role of antioxidant N-acetylcysteine (NAC) in intestinal epithelial cells against oxidative damage induced by H2O2. IPEC-J2 cells were cultured in DMEM-H with NAC and H2O2. After 2-day incubation, IPEC-J2 cells were collected for analysis of DNA synthesis, antioxidation capacity, mitochondrial respiration, and cell apoptosis. The results showed that H2O2 significantly decreased (P < 0.05) proliferation rate, mitochondrial respiration, and antioxidation capacity and increased cell apoptosis and the abundance of associated proteins, including cytochrome C, Bcl-XL, cleaved caspase-3, and total caspase-3. NAC supplementation remarkably increased (P < 0.05) proliferation rate, antioxidation capacity, and mitochondrial bioenergetics but decreased cell apoptosis. These findings indicate that NAC might rescue the intestinal injury induced by H2O2.

Glutamine promotes intestinal SIgA secretion through intestinal microbiota and IL-13.

SCOPE: Glutamine supplementation enhances secretory IgA (SIgA) production in the intestine, but the mechanism is largely unknown. We examined this issue using a mouse model. METHODS AND RESULTS: In mouse model, glutamine supplementation increased both SIgA abundance in intestinal luminal contents and IgA(+) plasma cell numbers in the mouse ileum, and decreased bacterial loads in mouse mesenteric lymph nodes. Glutamine supplementation increased mouse ileal expression of cytokines associated with T cell-dependent and T cell-independent pathways of SIgA induction, including IL-5, IL-6, IL-13, transforming growth factor (TGF-ß), a proliferation-inducing ligand (APRIL), B cell-activating factor (BAFF), vasoactive intestinal peptide (VIP) receptor, and retinal dehydrogenases. Injecting an IL-13 antibody during glutamine supplementation reduced J-chain expression in the mouse ileum. Glutamine supplementation increased bacterial invasion into the mouse ileal wall, while disrupting the mouse intestinal microbiota abrogated the influence of glutamine supplementation on SIgA secretion. CONCLUSION: Glutamine supplementation appears to enhance SIgA secretion in the mouse intestine through the intestinal microbiota and subsequently through T cell-dependent and T cell-independent pathways.

Pyrrolidine Dithiocarbamate Inhibits NF-KappaB Activation and Upregulates the Expression of Gpx1, Gpx4, Occludin, and ZO-1 in DSS-Induced Colitis.

Inflammatory bowel disease (IBD) correlates with oxidative stress, inflammation, and alteration in several signal pathways, including nuclear transcription factor-kappaB (NF-κB). Pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB, has been widely demonstrated to exhibit an antioxidant and anti-inflammatory function. This study aimed to test the hypothesis that NF-κB inhibitor PDTC confers a beneficial role in a colitis model induced by dextran sodium sulfate (DSS) in mouse. The results showed that DSS decreased daily weight gain, induced colonic inflammation, suppressed the expression of antioxidant enzymes and tight junctions, and activated NF-κB and nuclear factor erythroid 2-related factor 2/Kelch-like ECH-associated protein 1 (Nrf2/Keap1) signaling pathways. PDTC significantly upregulated (P < 0.05) Gpx1, Gpx4, occludin, and ZO-1 expressions in the DSS-induced colitis model. Meanwhile, PDTC reversed (P < 0.05) the activation of NF-κB signal pathway caused by DSS treatment. In conclusion, PDTC could serve as an adjuvant therapy for the patient with IBD.

An NMR-based metabolomic approach to investigate the effects of supplementation with glutamic acid in piglets challenged with deoxynivalenol.

Deoxynivalenol (DON) has various toxicological effects in humans and pigs that result from the ingestion of contaminated cereal products. This study was conducted to investigate the protective effects of dietary supplementation with glutamic acid on piglets challenged with DON. A total of 20 piglets weaned at 28 d of age were randomly assigned to receive 1 of 4 treatments (5 piglets/treatment): 1) basal diet, negative control (NC); 2) basal diet +4 mg/kg DON (DON); 3) basal diet +2% (g/g) glutamic acid (GLU); 4) basal diet +4 mg/kg DON +2% glutamic acid (DG). A 7-d adaptation period was followed by 30 days of treatment. A metabolite analysis using nuclear magnetic resonance spectroscopy (1H-NMR)-based metabolomic technology and the determination of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities for plasma, as well as the activity of Caspase-3 and the proliferation of epithelial cells were conducted. The results showed that contents of low-density lipoprotein, alanine, arginine, acetate, glycoprotein, trimethylamine-N-oxide (TMAO), glycine, lactate, and urea, as well as the glutamate/creatinine ratio were higher but high-density lipoprotein, proline, citrate, choline, unsaturated lipids and fumarate were lower in piglets of DON treatment than that of NC treatment (P<0.05). Compared with DON treatment, dietary supplementation with glutamic acid increased the plasma concentrations of proline, citrate, creatinine, unsaturated lipids, and fumarate, and decreased the concentrations of alanine, glycoprotein, TMAO, glycine, and lactate, as well as the glutamate/creatinine ratio (P<0.05). Addition glutamic acid to DON treatment increased the plasma activities of SOD and GSH-Px and the proliferating cell nuclear antigen (PCNA) labeling indexes for the jejunum and ileum (P<0.05). These novel findings indicate that glutamic acid has the potential to repair the injuries associated with oxidative stress as well as the disturbances of energy and amino acid metabolism induced by DON.

Dietary glutamate supplementation ameliorates mycotoxin-induced abnormalities in the intestinal structure and expression of amino acid transporters in young pigs.

The purpose of this study was to investigate the hypothesis that dietary supplementation with glutamic acid has beneficial effects on growth performance, antioxidant system, intestinal morphology, serum amino acid profile and the gene expression of intestinal amino acid transporters in growing swine fed mold-contaminated feed. Fifteen pigs (Landrace×Large White) with a mean body weight (BW) of 55 kg were randomly divided into control group (basal feed), mycotoxin group (contaminated feed) and glutamate group (2% glutamate+contaminated feed). Compared with control group, mold-contaminated feed decreased average daily gain (ADG) and increased feed conversion rate (FCR). Meanwhile, fed mold-contaminated feed impaired anti-oxidative system and intestinal morphology, as well as modified the serum amino acid profile in growing pigs. However, supplementation with glutamate exhibited potential positive effects on growth performance of pigs fed mold-contaminated feed, ameliorated the imbalance antioxidant system and abnormalities of intestinal structure caused by mycotoxins. In addition, dietary glutamate supplementation to some extent restored changed serum amino acid profile caused by mold-contaminated feed. In conclusion, glutamic acid may be act as a nutritional regulating factor to ameliorate the adverse effects induced by mycotoxins.

Therapeutic effects of glutamic acid in piglets challenged with deoxynivalenol.

The mycotoxin deoxynivalenol (DON), one of the most common food contaminants, primarily targets the gastrointestinal tract to affect animal and human health. This study was conducted to examine the protective function of glutamic acid on intestinal injury and oxidative stress caused by DON in piglets. Twenty-eight piglets were assigned randomly into 4 dietary treatments (7 pigs/treatment): 1) uncontaminated control diet (NC), 2) NC+DON at 4 mg/kg (DON), 3) NC+2% glutamic acid (GLU), and 4) NC+2% glutamic acid + DON at 4 mg/kg (DG). At day 15, 30 and 37, blood samples were collected to determine serum concentrations of CAT (catalase), T-AOC (total antioxidant capacity), H2O2 (hydrogen peroxide), NO (nitric oxide), MDA (maleic dialdehyde), DAO (diamine oxidase) and D-lactate. Intestinal morphology, and the activation of Akt/mTOR/4EBP1 signal pathway, as well as the concentrations of H2O2, MDA, and DAO in kidney, liver and small intestine, were analyzed at day 37. Results showed that DON significantly (P<0.05) induced oxidative stress in piglets, while this stress was remarkably reduced with glutamic acid supplementation according to the change of oxidative parameters in blood and tissues. Meanwhile, DON caused obvious intestinal injury from microscopic observations and permeability indicators, which was alleviated by glutamic acid supplementation. Moreover, the inhibition of DON on Akt/mTOR/4EBP1 signal pathway was reduced by glutamic acid supplementation. Collectively, these data suggest that glutamic acid may be a useful nutritional regulator for DON-induced damage manifested as oxidative stress, intestinal injury and signaling inhibition.

Serum amino acids profile and the beneficial effects of L-arginine or L-glutamine supplementation in dextran sulfate sodium colitis.

This study was conducted to investigate serum amino acids profile in dextran sulfate sodium (DSS)-induced colitis, and impacts of graded dose of arginine or glutamine supplementation on the colitis. Using DSS-induced colitis model, which is similar to human ulcerative colitis, we determined serum profile of amino acids at day 3, 7, 10 and 12 (5 days post DSS treatment). Meanwhile, effects of graded dose of arginine (0.4%, 0.8%, and 1.5%) or glutamine (0.5%, 1.0% and 2.0%) supplementation on clinical parameters, serum amino acids, colonic tight junction proteins, colonic anti-oxidative indicators [catalase, total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px)], colonic pro-inflammatory cytokines [interleukin-1 beta (IL-1ß), IL-6, IL-17 and tumor necrosis factor alpha (TNF-α)] in DSS-induced colitis were fully analyzed at day 7 and 12. Additionally, the activation of signal transduction pathways, including nuclear factor kappa B (NF-κB), mitogen-activated protein kinases (MAPK), phosphoinositide-3-kinases (PI3K)/PI3K-protein kinase B (Akt), and myosin light chain kinase (MLCK)-myosin light chain (MLC20), were analyzed using immunoblotting. Serum amino acids analysis showed that DSS treatment changed the serum contents of amino acids, such as Trp, Glu, and Gln (P<0.05). Dietary arginine or glutamine supplementation had significant (P<0.05) influence on the clinical and biochemical parameters (T-SOD, IL-17 and TNF-α) in colitis model. These results were associated with colonic NF-κB, PI3K-Akt and MLCK signaling pathways. In conclusion, arginine or glutamine could be a potential therapy for intestinal inflammatory diseases.

Dietary supplementation with proline confers a positive effect in both porcine circovirus-infected pregnant and non-pregnant mice.

Porcine circovirus type 2 (PCV2) is associated with various diseases that impose a significant economic burden on the swine industry. We hypothesised that nutritional supplementation with proline to enhance the immune response might be a useful prophylactic measure against PCV2 infection. To test this hypothesis, in the present study, we measured clinical data, including blood parameters, serum cytokine profile, PCV2 virus load in organs and serum, and microscopic lesions in the lung, liver and spleen, in both PCV2-infected pregnant and non-pregnant mice. Dietary supplementation with proline had no effect (P>0·05) on abortion rates in PCV2-infected pregnant mice, although a numerically lower abortion rate (22·2 v. 44·4%) was observed compared with the control. Dietary supplementation with proline significantly increased serum C-reactive protein levels (P= 0·03) in PCV2-infected pregnant mice, and increased serum TNF-α levels (P= 0·01), leucocytes (P< 0·05), lymphocytes (P< 0·05) and neutrophilic granulocytes (P< 0·05) in PCV2-infected non-pregnant mice. Meanwhile, dietary proline significantly (P< 0·05) decreased the PCV2 virus load in the lung. Furthermore, mice in the dietary proline group showed a significant (P< 0·01) decrease in microscopic lesion scores in the lung, liver and spleen compared with those in the alanine group. Collectively, dietary proline supplementation confers a functional role in PCV2-infected mice.

Dietary L-glutamine supplementation improves pregnancy outcome in mice infected with type-2 porcine circovirus.

Porcine circovirus type 2 (PCV2) causes reproductive failure in swine. As glutamine can enhance immune function in animals, this study was conducted with mice to test the hypothesis that dietary glutamine supplementation will improve pregnancy outcome in PCV2-infected dams. Beginning on day 0 of gestation, mice were fed a standard diet supplemented with 1.0% L-glutamine or 1.22% L-alanine (isonitrogenous control). All mice were infected with PCV2 (2000 TCID50) on day 10 of gestation. On day 17 of gestation, six mice from each group were euthanized to obtain maternal tissues and fetuses for hematology and histopathology tests. The remaining mice continued to receive their respective diets supplemented with 1.0% L-glutamine or 1.22% L-alanine through lactation. The PCV2 virus was present in maternal samples (serum and lung) of most mice in the control group but was not detected in the glutamine-supplemented mice. Dietary glutamine supplementation reduced abortion, decreased fetal deaths, and enhanced neonatal survival. The glutamine treatment also reduced concentrations of interleukin-6, while increasing concentrations of tumor necrosis factor-α and C-reactive protein, in the maternal serum of mice. Furthermore, glutamine supplementation attenuated microscopic lesions in maternal tissues (lung, spleen, and liver). Collectively, these results indicate that dietary glutamine supplementation is beneficial for ameliorating reproductive failure in virus-infected mice. The findings support the notion that gestating dams require adequate amounts of dietary glutamine for the optimal survival and growth of embryos, fetuses, and neonates, and have important implications for nutritional support of mammals (including swine and humans) during gestation and lactation.