Eugenol Possesses Colitis Protective Effects: Impacts on the TLR4/MyD88/NF-[Formula: see text]B Pathway, Intestinal Epithelial Barrier, and Macrophage Polarization.

Eugenol (EU) has been shown to ameliorate experimental colitis due to its anti-oxidant and anti-inflammatory bioactivities. In this study, DSS-induced acute colitis was established and applied to clarify the regulation efficacy of EU on intestinal barrier impairment and macrophage polarization imbalance along with the inflammatory response. Besides, the adjusting effect of EU on macrophages was further investigated in vitro. The results confirmed that EU intervention alleviated DSS-induced colitis through methods such as restraining weight loss and colonic shortening and decreasing DAI scores. Microscopic observation manifested that EU maintained the intestinal barrier integrity in line with the mucus barrier and tight junction protection. Furthermore, EU intervention significantly suppressed the activation of TLR4/MyD88/NF-[Formula: see text]B signaling pathways and pro-inflammatory cytokines gene expressions, while enhancing the expressions of anti-inflammatory cytokines. Simultaneously, WB and FCM analyses of the CD86 and CD206 showed that EU could regulate the DSS-induced macrophage polarization imbalance. Overall, our data further elucidated the mechanism of EU's defensive effect on experimental colitis, which is relevant to the protective efficacy of intestinal barriers, inhibition of oxidative stress and excessive inflammatory response, and reprogramming of macrophage polarization. Hence, this study may facilitate a better understanding of the protective action of the EU against UC.

Therapeutic effect of a self-made herbal formula on a multi-drug resistant Eimeria tenella isolate infection in broiler chickens.

In-feed prophylactic chemotherapy is widely considered the mainstay of avian coccidiosis control, while serious drug resistance strictly restricts its application. Confronted with the urgent need for an alternative strategy, a traditional Chinese medicine formula (TCMF) was developed. Meanwhile, its potential to iron out complicated clinical coccidiosis was scrutinized in vivo with a field-isolated multi-drug resistant Eimeria tenella (E. tenella) isolate. Birds were inoculated with 5 × 104 sporulated oocysts and administrated with TCMF supplementation in water from 72 h post-infection to the end of the experiment, diclazuril (DIC) was set as a positive control. As a result, TCMF intervention reduced oocyst shedding, cecal lesion and mortality, and enhanced body weight gain. According to the above, anticoccidial index (ACI) was calculated and TCMF exerted a moderate anticoccidial activity. Besides, macroscopic, histopathological, and ultrastructural observations revealed the safeguarding effects of TCMF on E. tenella-induced cecal injury. Following, TCMF treatment presented an obvious inhibition effect on E. tenella caused oxidative stress and inflammatory response. Moreover, TCMF supplementation restored the cecal flora abundance and diversity, reduced the colonization of harmful bacteria, and increased the probiotics abundance. In conclusion, TCMF exhibited a moderate anticoccidial effect along with alleviating E. tenella-induced cecal injury, redox imbalance, and inflammatory response which may be associated with the microflora modulatory effect.

Acupuncture in the Treatment of Amyotrophic Lateral Sclerosis: A Research Progress in Clinical Trials.

Background: Acupuncture, a complementary and alternative medicine (CAM) modality, shows promise as an integrative therapy for Amyotrophic Lateral Sclerosis (ALS) due to the chronic nature of the disease and its persistent symptoms. Many patients turn to CAM for ALS treatment. Objective: This review assesses acupuncture's efficacy in treating Amyotrophic Lateral Sclerosis. Methods: We searched China National Knowledge Network (CNKI) and PubMed databases for Chinese and English articles, including clinical trials, case studies, cohorts, and randomized controlled trials. The search, performed on March 31, 2023, encompassed literature published up to that date. Keywords used in titles and abstracts were (acupuncture) OR (electro-acupuncture)) AND (Amyotrophic Lateral Sclerosis). Results: Among the 45 articles studied, 34 were included in this research. Acupuncture's benefits primarily lie in neuro-immune system regulation, enhanced quality of life, reduced fatigue, disease progression delay, and fewer relapses. Conclusions: Recent clinical trials highlight the potential of traditional Chinese acupuncture in improving Amyotrophic Lateral Sclerosis symptoms (e.g., fatigue, neural functional deficits) and curtailing relapses. Consequently, acupuncture holds promise as an integrative therapy for ALS patients.

Electro-nape-acupuncture regulates the differentiation of microglia through PD-1/PD-L1 reducing secondary brain injury in acute phase intracerebral hemorrhage rats.

INTRODUCTION: This study aimed to investigate the effect of electro-nape-acupuncture (ENA) on the differentiation of microglia and the secondary brain injury in rats with acute-phase intracerebral hemorrhage (ICH) through the programmed cell death protein-1/ligand-1 (PD-1/PD-L1) pathway. METHODS: A total of 27 male Sprague-Dawley rats were randomly divided into three groups: sham group, ICH group, and ENA group. The autologous blood infusion intracerebral hemorrhage model was used to study the effects of ENA by administering electroacupuncture at GB20 (Fengchi) and Jiaji (EX-B2) acupoints on 24 h after the modeling, once per day for 3 days. The neurological function damage, hematoma lesion, and inflammatory cell infiltration were measured by the beam walking test and hematoxylin-eosin staining. The expression of PD-1, PD-L1, CD86, CD206, and related cytokines around the hematoma was measured by western blot, quantitative reverse transcription polymerase chain reaction, and immunofluorescence. RESULTS: The ICH group had significant neurological deficits (p < .001), hematoma lesions, and inflammatory cell infiltration. The levels of CD86 protein, inflammatory factors tumor necrosis factors (TNF)-α, interleukin (IL)-1ß, and IL-6 were increased (p < .001), while CD206 protein was reduced (p < .01), and the number of CD86+ /CD11b+ cells was also increased (p < .001) compared to the sham group. However, after ENA intervention, there was a significant reduction in neurological function damage (p < .05), infiltration of inflammatory cells, and the expression levels of CD86+ /CD11b+ cells (p < .05), resulting in the increased expression of PD-1 protein and differentiation of M2 phenotype significantly (p < .001). CONCLUSION: The study concludes that ENA could reduce neurological function damage, inhibit the expression of pro-inflammatory cytokines, and improve the infiltration of inflammatory cells to improve secondary brain injury in acute-phase intracerebral hemorrhage rats. These effects could be related to the increased expression of PD-1 around the lesion, promoting the differentiation of microglia from M1 to M2 phenotype.

Electroacupuncture regulates microglial polarization via inhibiting NF-κB/COX2 pathway following traumatic brain injury.

BACKGROUND: Neuroinflammation and oxidative stress are important pathological mechanisms following traumatic brain injury (TBI). The NF-κB/COX2 pathway regulates neuroinflammation and oxidative damage, while microglia also play an important role in neuroinflammation. Since NF-κB is involved in microglial polarization, targeting this pathway and microglial polarization is a critical component of TBI treatment. Currently, electroacupuncture (EA) is widely used to treat various symptoms after TBI, but the mechanisms of EA remain poorly understood. Additionally, the optimal frequency of EA remains unclear, which affects its efficacy. This study focuses on exploring the optimal frequency parameters of EA on TBI and investigating the underlying mechanisms of EA through NF-κB/COX2 pathway and microglial polarization. METHODS: The study was divided into two parts. In Experiment 1, 42 Sprague Dawley (SD) rats were induced and randomly divided into seven groups (n = 6). Except for the sham group, all rats underwent controlled cortical impact (CCI) to establish TBI model. Four EA groups (with different frequencies) and manual acupuncture (without current stimulation) received stimulation on the acupoints of Shuigou (GV26), Fengchi (GB20) and Neiguan (PC6) once a day for 7 days. The neurological function was assessed by modified Neurological Severity Scores (mNSS), and the rats' memory and learning were examined by the Morris water maze (MWM). SOD, MDA, and GSH-Px were detected to evaluate the levels of oxidative stress. The levels of IL-1ß, IL-6, and TNF-α were evaluated by Enzyme Linked Immunosorbent Assay (ELISA). Detection of the above indicators indicated a treatment group that exerted the strongest neuroprotection against TBI, we then conducted Experiment 2 using this screened acupuncture treatment to investigate the mechanism of acupuncture. 48 rats were randomly divided into four groups (n = 12): sham, TBI model, acupuncture and PDTC (NF-κB inhibitor). Evaluations of mNSS, MWM test, SOD, MDA, GSH-Px, IL-1ß, IL-6, TNF-α, and IL-10 were the same as in Experiment 1. Western blot was applied for detecting the expression levels of NF-κB, p-NF-κB, COX2, and Arg-1. TUNEL was used to examine neuronal apoptosis. Brain structure was observed by H&E. Iba-1, COX2, and Arg-1 were investigated by immunofluorescence staining. RESULTS: EA with frequency of 2/100 Hz markedly improved neuronal and cognitive function as compared to the other treatment groups. Moreover, it downregulated the expression of MDA, IL-6, IL-1ß, and TNF-α and upregulated the levels of SOD and GSH-Px. In addition, Both EA with 2/100 Hz and PDTC reduced the levels of p-NF-κB, COX2 and M1 markers (COX2, IL-6, IL-1ß, TNF-α) and increased the levels of M2 markers (Arg-1, IL-10). Moreover, they had similar effects on reducing inflammation, oxidative stress and apoptosis, and improving neuronal and cognitive function. CONCLUSIONS: The collective findings strongly suggest that EA with 2/100 Hz can improve neurologic function by suppressing neuroinflammation, oxidative stress and apoptosis. Additionally, we confirm that EA promotes microglial polarization towards the M2 phenotype through the suppression of NF-κB/COX2 pathway, thus exerting neuroprotective effects after TBI.

Acupuncture for acute migraine attacks in adults: a systematic review and meta-analysis.

Objective To evaluate the effect and safety of acupuncture for acute migraine attacks in adults. DESIGN AND SETTING: We searched PubMed, MEDLINE(OVID), Chinese Biomedical Literature Database, China National Knowledge Infrastructure, Chinese Science and Technology Periodical Database and Wanfang database from inception to 15 July 2022. We included randomised controlled trials (RCTs) published in Chinese and English comparing acupuncture alone against sham acupuncture/placebo/no treatment/pharmacological therapy or comparing acupuncture plus pharmacological therapy against the same pharmacological therapy. Results were reported as risk ratios (RRs) for dichotomous outcomes or mean differences (MDs) for continuous outcomes, with 95% CIs. Risk of bias was assessed with the Cochrane tool and the certainty of the evidence (CoE) with GRADE. : Main outcome measures : a) The rate of headache freedom (pain score=0) at 2h after the treatment; b) the rate of headache relief (at least 50% reduction of pain score); c) headache intensity at 2h after the treatment(study data from scales measuring pain intensity, including visual analogue scale, numerical rating scale) d) the improvement of headache intensity at 2h after the treatment; e) the improvement values of migraine-associated symptoms; f) adverse events. RESULTS: We included 21 RCTs from 15 studies with 1926 participants comparing acupuncture against other interventions. Comparing to sham acupuncture or placebo, acupuncture may result in an increase in the rate of headache freedom (RR 6.03, 95% CI 1.62 to 22.41, 180 participants, 2 studies, I2=0%, low CoE) and the improvement of headache intensity (MD 0.51, 95% CI 0.16 to 0.85, 375 participants, 5 studies, I2=13%, moderate CoE) at 2 hours after treatment. It also may result on a higher rate of headache relief (RR 2.29, 95% CI 1.16 to 4.49, 179 participants,3 studies, I2=74%, very low CoE) and greater improvement of migraine-associated symptoms (MD 0.97, 95% CI 0.33 to 1.61, 90 participants, 2 studies, I2=0%, very low CoE) at 2 hours after treatment but the evidence is very uncertain. Meanwhile the analysis indicates acupuncture probably results in little to no difference in adverse events compared with sham acupuncture (RR 1.53, 95% CI 0.82 to 2.87, 884 participants, 10 studies, I2=0%, moderate CoE). In acupuncture plus pharmacological intervention versus the same pharmacological intervention, acupuncture plus pharmacological therapy may result in little to no difference in the rate of headache freedom (RR 1.55, 95% CI 0.99 to 2.42, 94 participants, 2 studies, I2=0%, low CoE), the rate of headache relief (RR 1.20, 95% CI 0.91 to 1.57, 94 participants, 2 studies, I2=0%, low CoE) at 2 hours after treatment and adverse events(RR 1.48, 95% CI 0.25 to 8.92, 94 participants, 2 studies, I2=0%, low CoE). However, it may result in a reduction in headache intensity (MD -1.05, 95% CI -1.49 to -0.62, 129 participants, 2 studies, I2=0%, low CoE) and an increase in the improvement of headache intensity (MD 1.18, 95% CI 0.41 to 1.95, 94 participants, 2 studies, I2=0%, low CoE) at 2 hours after treatment compared with pharmacological therapy only. In comparison to pharmacological intervention, acupuncture may result in little to no difference in the rate of headache freedom (RR 0.95, 95% CI 0.59 to 1.52, 294 participants, 4 studies, I2=22%, low CoE), the rate of headache relief (RR 0.95, 95% CI 0.80 to 1.14, 206 participants, 3 studies. I2=0%, low CoE) at 2 hours and adverse events (RR 0.65, 95% CI 0.35 to 1.22, 294 participants, 4 studies, I2=0%, low CoE) after treatment. The evidence is very uncertain about the effect of acupuncture on the headache intensity (MD -0.07, 95% CI -1.11 to 0.98, 641 participants, 5 studies, I2=98%, very low CoE) and the improvement of headache intensity (MD -0.32, 95% CI -1.07 to 0.42, 95 participants, 2 studies, I2=0%, very low CoE) at 2 hours after treatment compared with pharmacological intervention. CONCLUSION: The body of evidence suggests that acupuncture may be more effective than sham acupuncture in the treatment of migraine. Acupuncture may also be as effective as pharmacological therapy. However, the certainty evidence across outcomes was low to very low and new high-quality studies can provide more clarity. PROSPERO REGISTRATION NUMBER: CRD42014013352.

Acupuncture for the Treatment of Migraine: An Overview of Systematic Reviews.

PURPOSE OF REVIEW: Over the past few years, there have been many systematic reviews (SRs) and meta-analyses (MAs) used to assess the safety and efficacy of acupuncture in the treatment of migraine. Our aim is to assess the methodological and reporting quality of SRs/MAs and make an evaluation about the available evidence of the safety and efficacy of acupuncture in the treatment of migraine. RECENT FINDINGS: Migraine is one of the most common primary headaches, with various symptoms, threatening human health. Acupuncture is a specific treatment of Traditional Chinese Medicine (TCM), and a validated non-pharmaceutical option as well, widely used in the treatment of migraine, and has gained remarkable therapeutic effect. An overview can evaluate research methods and evidence for SRs/MAs in evidence-based medicine, although there is much relevant research, it's still a hard work to synthesis all the evidence or to make robust conclusions, the variation in the methodological and quality evidence in such SRs/MAs play a significant role. For this overview, we searched six electronic databases from inception until 8 September 2022, and without languages restrictions, the results showed that, acupuncture as a more safety and more convenient therapeutic, and it has been shown to be effective in the treatment of migraine, that is worthy of clinical promotion. However, there are also some limitations because of the low-quality evidence of most of the studies. In conclusion, most included SRs/MAs suggested that acupuncture was more effective than the control group in the treatment of migraine. However, the quality evidence of most of the studies still needs to be improved.

Eugenol alleviates Salmonella Typhimurium-infected cecal injury by modulating cecal flora and tight junctions accompanied by suppressing inflammation.

BACKGROUND: Salmonella enterica serovar Typhimurium (ST) mainly exists in poultry and poultry related products, which are common sources of human salmonellosis. So, ST is an important zoonotic pathogen that threatens public health and safety. Eugenol has been noted for its antibacterial and anti-inflammatory properties, and it is expected to develop into an antibacterial therapy in vivo. METHODS: Herein, the effects of eugenol against ST infection in terms of intestinal flora, cecal tight junction, and cecal inflammation in broilers was evaluated in this study. RESULTS: The results showed that oral eugenol pretreatment obviously relieved the histopathological changes and ultrastructural injury of cecum caused by ST infection. Further analysis found that eugenol lessened the number of ST in the cecum, restrained Proteobacteria and Ruminococcus, and kept the ratio of Firmicutes to Bacteroidetes (F/B), improved the relative abundance of Alistipes compared to the infection control. Moreover, eugenol sustained the expression of ZO-1, claudin-1, and occluding tight junction proteins, reduced the mRNA levels of myeloid differentiation factor 88 (MyD88), toll-like receptor-4 (TLR4) and inducible nitric oxide synthesis (iNOS), suppressed the phosphorylation of p65 and IκBα in the NF-κB signaling pathway, as well as the mRNA levels of TNF-α, IL-1ß, IL-2, and IL-18 in the cecum tissue. CONCLUSION: Taken together, eugenol could protect broilers against ST infection via maintaining intestinal flora, tight junctions involved in mucosal barrier function, then accordingly limiting the excessive inflammation, finally contributed to improving relative weight gains and survival rates of broilers.

Bactericidal activity of gallic acid against multi-drug resistance Escherichia coli.

The continuous emergence of multidrug-resistant (MDR) bacteria has posed an increasingly serious public health threat which urges people to develop some alternatives. Gallic acid (GA) is a natural ingredient in many traditional Chinese medicines, which has many biological activities, such as antibacterial, and antiseptic. Here, clinical isolates of MDR Escherichia coli (E. coli) were used to evaluate the antibacterial effect of GA and the underlying mechanism. The results revealed that GA exerted bactericidal activity and inhibited the formation of bacterial biofilm. GA enhanced the activities of ceftiofur sodium or tetracycline against E. coli, and facilitated antibiotic accumulation in bacteria. Further analysis of morphological alterations and efflux pump gene expressions confirmed that GA damaged outer and inner membranes, and suppressed the mRNA expressions of acrA, acrB, tolC, acrD and acrF involved in membrane permeability. In addition, GA showed protective effects against bacterial infection and improved the survival rates of Galleria mellonella and BALB/c mice. These data highlight a better understanding of GA against bacteria and provide an alternative strategy for MDR bacterial infection.

Emodin from Aloe inhibits Swine acute diarrhea syndrome coronavirus in cell culture.

Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an emerging swine enteropathogenic coronavirus that causes severe diarrhea in neonatal piglets, leading to serious economic losses to the pig industries. At present, there are no effective control measures for SADS, making an urgent need to exploit effective antiviral therapies. Here, we confirmed that Aloe extract (Ae) can strongly inhibit SADS-CoV in Vero and IPI-FX cells in vitro. Furthermore, we detected that Emodin from Ae had anti-SADS-CoV activity in cells but did not impair SADS-CoV infectivity directly. The time-of-addition assay showed that Emodin inhibits SADS-CoV infection at the whole stages of the viral replication cycle. Notably, we found that Emodin can significantly reduce virus particles attaching to the cell surface and induce TLR3 (p < 0.001), IFN-λ3 (p < 0.01), and ISG15 (p < 0.01) expressions in IPI-FX cells, indicating that the anti-SADS-CoV activity of Emodin might be due to blocking viral attachment and the activation of TLR3-IFN-λ3-ISG15 signaling axis. These results suggest that Emodin has the potential value for the development of anti-SADS-CoV drugs.

Acupuncture for oculomotor nerve palsy:A systematic review and meta-analysis.

BACKGROUND: To evaluate the safety and efficacy of acupuncture in the treatment of oculomotor nerve palsy (ONP). METHODS: The following database will be required from PubMed, Cochrane Library, Medline, Chinese Biomedical Literature Database, China National Knowledge Infrastructure (CNKI), Wanfang data. Randomized controlled trials (RCTs) comparing acupuncture alone versus no treatment/another active therapy/sham acupuncture or comparing acupuncture with another active therapy versus the same active therapy were included. Meta-analysis was conducted according to the 2020 PRISMA guidelines. Data was analyzed using RevMan 5.4 software. Results were reported as risk ratios (RR) for dichotomous outcomes or mean differences (MD) for continuous outcomes, with 95 % confidence intervals (CI). The Cochrane risk of bias tool was used to access the methodological quality of the trails. RESULTS: Eighteen RCTs with 1150 participants comparing acupuncture versus other therapies were included. The results showed a significant differences in the clinical efficiency rate (RR = 1.30, 95 %CI = 1.23-1.37, P < 0.001), scores of diplopia (MD = - 0.78, 95 %CI = - 1.39 to - 0.77, P < 0.001), palpebral fissure size (MD = 1.04, 95 %CI = 0.41-1.68, P = 0.001), the pupil diameter (MD = - 0.56, 95 %CI = - 0.70 to - 0.42, P < 0.001), quality of life (MD = 8.96, 95 %CI = 6.79-11.13, P < 0.001) between the experiment and control groups. However, there were no significant differences in the adverse effects (RR = 0.52, 95 %CI = 0.22-1.22, P = 0.13). The quality of the evidence test by GRADE was low or very low. CONCLUSION: Most included studies suggested that acupuncture was more effective than the control group in the treatment of ONP. However, the quality evidence of most of the studies was low and most of them were performed in China.

A comparative study on roles of natural killer T cells in two diet-induced non-alcoholic steatohepatitis-related fibrosis in mice.

BACKGROUND: Immune responses are important in the progression of non-alcoholic fatty liver disease (NAFLD). Natural killer T (NKT) cells are main components of the innate immune system that modulate immunity. However, the role of NKT cells in NAFLD remains controversial. OBJECTIVE: We aimed to investigate the role of NKT cells in non-alcoholic steatohepatitis (NASH)-related fibrosis in fast food diet (FFD)- and methionine choline-deficient (MCD) diet-induced mouse models. METHODS: Hepatic NKT cells were analysed in wild-type (WT) and CD1d-/- mice fed FFD or MCD diets. Hepatic pathology, cytokine profiles and liver fibrosis were evaluated. Furthermore, the effect of chronic administration of α-galactosylceramide (α-GalCer) on liver fibrosis was investigated in both FFD- and MCD-treated mice. RESULTS: FFD induced a significant depletion of hepatic NKT cells, thus leading to mild to moderate NASH and early-stage fibrosis, while mice fed MCD diets developed severe liver inflammation and progressive fibrosis without a significant change in hepatic NKT cell abundance. FFD induced a similar liver fibrogenic response in CD1d-/- and WT mice, while MCD induced a higher hepatic mRNA expression of Col1α1 and TIMP1 as well as relative fibrosis density in CD1d-/- mice than WT mice (31.8 vs. 16.3, p = .039; 40.0 vs. 22.6, p = .019; 2.24 vs. 1.59, p = .036). Chronic administration of α-GalCer induced a higher hepatic mRNA expression of TIMP1 in MCD-treated mice than controls (36.7 vs. 14.9, p = .005). CONCLUSION: NKT cells have protective roles in NAFLD as the disease progresses. During diet-induced steatosis, mild to moderate NASH and the early stage of fibrosis, hepatic NKT cells are relatively depleted, leading to a proinflammatory status. In severe NASH and the advanced stage of liver fibrosis, NKT cells play a role in inhibiting the NASH-related fibrogenic response. Chronic administration of α-GalCer induces NKT cell anergy and tolerance, which may play a role in promoting the liver fibrogenic response.

Rapamycin Alleviates 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis through Autophagy Induction and NF-κB Pathway Inhibition in Mice.

Background: Recent genetic studies indicated that variants of autophagy genes were associated with the predisposition of Crohn's disease (CD). The autophagy deficiency may affect the innate and adaptive immunity, which is related to persistent and excessive inflammation of the bowel. However, it remains unclear how autophagy modulates the expression of immune response regulator NF-κB and proinflammatory cytokine TNF-α in CD. Aim: We aimed to investigate the role of rapamycin on the expression of NF-κB p65 and TNF-α in 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced mouse colitis and lipopolysaccharide (LPS)-induced HT-29 cells. Methods: TNBS-induced colitis mice were treated with saline or rapamycin, and the disease activity index (DAI) and histological scores of colonic mucosa were evaluated. The expressions of p65, ATG16L1 and LC3 were detected by western blot and immunohistochemistry staining. The monodansylcadaverine (MDC) staining and transmission electron microscopy were developed to study the autophagy in LPS-induced HT-29 cells. Expression of TNF-α from colon tissue and HT-29 cells were detected by ELISA. The expressions of p65, ATG16L1 and LC3 in active CD patients were also investigated. Results: Significantly more autophagosomes were observed in rapamycin-treated cells than in controls. Rapamycin remarkably upregulated the expression of ATG16L1 and LC3II, inhibited p65 nucleus translocation and secretion of TNF-α both in vivo and in vitro. The expression of both ATG16L1 and LC3II increased in mild to moderate CD specimens, while no significant difference was noted between severe CD and normal controls. The expression of p65 increased notably in severe CD compared to those in mild to moderate patients. Conclusions: In LPS-treated HT-29 cells and TNBS-induced colitis, p65 is overexpressed, which results in exaggerated secretion of TNF-α and induce or worsen the inflammation in the bowel. Rapamycin protects against colitis through induction of autophagy, thus inhibiting the activation of NF-κB pathway and secretion of TNF-α.

Poly(A)-Binding Protein Cytoplasmic 1 Inhibits Porcine Epidemic Diarrhea Virus Replication by Interacting with Nucleocapsid Protein.

Porcine epidemic diarrhea virus (PEDV) is the etiological agent of porcine epidemic diarrhea (PED) characterized by vomit, watery diarrhea, dehydration and high mortality. Outbreaks of highly pathogenic variant strains of PEDV have resulted in extreme economic losses to the swine industry all over the world. The study of host-virus interaction can help to better understand the viral pathogenicity. Many studies have shown that poly(A)-binding proteins are involved in the replication process of various viruses. Here, we found that the infection of PEDV downregulated the expression of poly(A)-binding protein cytoplasmic 1 (PABPC1) at the later infection stage in Vero cells. The overexpression of PABPC1 inhibited the proliferation of PEDV at transcription and translation level, and siRNA-mediated depletion of PABPC1 promoted the replication of PEDV. Furthermore, mass spectrometry analysis and immunoprecipitation assay confirmed that PABPC1 interacted with the nucleocapsid (N) protein of PEDV. Confocal microscopy revealed the co-localizations of PABPC1 with N protein in the cytoplasm. Taken together, these results demonstrate the antiviral effect of PABPC1 against PEDV replication by interacting with N protein, which increases understanding of the interaction between PEDV and host.

The protective effect and potential mechanisms of eugenol against Salmonella in vivo and in vitro.

Salmonella enterica serovar Typhimurium (S. Typhimurium) continues to be a serious concern to the poultry industry as a bacterial foodborne zoonosis, which generally results in intestinal inflammation and barrier dysfunction or even death. Eugenol is a phenolic compound with various pharmacological activities involved antioxidant, anti-inflammatory, and antibacterial effects, which is expected to be an effective nonantibiotic therapy. The purpose of this study was to explore the protective effects of eugenol in the cellular and broiler models of S. Typhimurium infection and the possible underlying mechanisms. The results of animal infection showed that eugenol treatments enhanced the relative weight gains and survival rates of broilers with a reduction of the organ bacterial load and intestinal ultrastructural injury. Moreover, eugenol significantly inhibited the mRNA levels of myeloid differentiation factor 88 (MyD88) and toll-like receptor-4 (TLR4), then declined the phosphorylation of p65 and IκBα of NF-κB pathway and the expressions of inflammatory factors (TNF-α, IL-1ß, IL-2, and IL-18) in duodenum tissues, while maintained the expressions of intestinal tight junction proteins (ZO-1, claudin-1, occludin). Further experiments in vitro revealed that eugenol markedly inhibited the adhesion and invasion of S. Typhimurium to RAW264.7 or IEC-6 cells, then reduce bacterial multiplication in IEC-6 or DF-1 cells. In conclusion, eugenol could defend broilers from S. Typhimurium infection by stabilizing the intestinal mucosal barrier and relieving inflammatory response, as well as inhibiting bacterial adhesion and invasion to cells.

Development of an indirect ELISA for detecting swine acute diarrhoea syndrome coronavirus IgG antibodies based on a recombinant spike protein.

Swine acute diarrhoea syndrome coronavirus (SADS-CoV) is a newly identified swine enteropathogenic coronavirus that causes watery diarrhoea in neonatal piglets, leading to significant economic losses to the swine industry. Currently, there are no suitable serological methods to assess the infection of SADS-CoV and effectiveness of vaccines, making an urgent need to exploit effective enzyme-linked immunosorbent assay (ELISA) to compensate for this deficiency. Here, a recombinant plasmid that expresses the spike (S) protein of SADS-CoV fused to the Fc domain of human IgG was constructed to generate recombinant baculovirus and expressed in HEK 293F cells. The S-Fc protein was purified with protein G Resin, which retained reactivity with anti-human Fc and anti-SADS-CoV antibodies. The S-Fc protein was then used to develop an indirect ELISA (S-iELISA) and the reaction conditions of S-iELISA were optimized. As a result, the cut-off value was determined as 0.3711 by analyzing OD450nm values of 40 SADS-CoV-negative sera confirmed by immunofluorescence assay (IFA) and western blot. The coefficient of variation (CV) of 6 SADS-CoV-positive sera within and between runs of S-iELISA were both less than 10%. The cross-reactivity assays demonstrated that S-iELISA was non-cross-reactive with other swine viruses' sera. Furthermore, the overall coincidence rate between IFA and S-iELISA was 97.3% based on testing 111 clinical serum samples. Virus neutralization test with seven different OD450nm values of the sera showed that the OD450nm values tested by S-iELISA are positively correlated with the virus neutralization assay. Finally, a total of 300 pig field serum samples were tested by S-iELISA and commercial kits of other swine enteroviruses showed that the IgG-positive for SADS-CoV, TGEV, PDCoV and PEDV was 81.7, 54, 65.3 and 6%, respectively. The results suggest that this S-iELISA is specific, sensitive, repeatable and can be applied for the detection of the SADS-CoV infection in the swine industry.

Isolated hepatic tuberculosis associated with portal vein thrombosis and hepatitis B virus coinfection: A case report and review of the literature.

BACKGROUND: While tuberculosis (TB) itself is a common disease, isolated TB of the liver is a rare entity. Tubercular involvement of the liver is more commonly a part of a disseminated disease of the hepatic parenchyma. In contrast, isolated hepatic TB spread through the portal vein from the gastrointestinal tract is seldom encountered in clinical practice, with only a few sporadic cases and short series available in the current literature. Vascular complications, such as portal vein thrombosis (PVT), have rarely been reported previously. CASE SUMMARY: A 22-year-old man was hospitalized with complaints of a 3-mo history of fever and weight loss of approximately 10 kg. He had a 10-year hepatitis B virus (HBV) infection in his medical history. Contrast-enhanced computed tomography (CECT) confirmed hepatosplenomegaly, with hypodensity of the right lobe of the liver and 2.1 cm thrombosis of the right branch of the portal vein. A liver biopsy showed epithelioid granulomas with a background of caseating necrosis. Ziehl-Nelson staining showed acid-fast bacilli within the granulomas. The patient was diagnosed with isolated hepatic TB with PVT. Anti-TB therapy (ATT), including isoniazid, rifapentine, ethambutol, and pyrazinamide, was administered. Along with ATT, the patient was treated with entecavir as an antiviral medication against HBV and dabigatran as an anticoagulant. He remained asymptomatic, and follow-up sonography of the abdomen at 4 mo showed complete resolution of the PVT. CONCLUSION: Upon diagnosis of hepatic TB associated with PVT and HBV coinfection, ATT and anticoagulants should be initiated to prevent subsequent portal hypertension. Antiviral therapy against HBV should also be administered to prevent severe hepatic injury.

Emodin from Aloe Inhibits Porcine Reproductive and Respiratory Syndrome Virus via Toll-Like Receptor 3 Activation.

Porcine reproductive and respiratory syndrome virus (PRRSV) causes severe reproductive failure in sows and respiratory diseases in growing and finishing pigs and results in great economic losses to the swine industry. Although vaccines are available, PRRSV remains a major threat to the pig farms. Thus, there is an urgent need to develop antiviral drugs to compensate for vaccines. In this study, we report that Aloe extract (Ae) can strongly inhibit PRRSV in Marc-145 cells and porcine alveolar macrophages lines (iPAMs) in vitro. Furthermore, we identified a novel anti-PRRSV molecule, Emodin, from Ae by high-performance liquid chromatography (HPLC). Emodin exerted its inhibitory effect through targeting the whole stages of PRRSV infectious cycle. Moreover, we also found that Emodin can inactivate PRRSV particles directly. Notably, we confirmed that Emodin was able to significantly induce Toll-like receptor 3 (TLR3) (p < 0.01), IFN-α (p < 0.05) and IFN-ß expression in iPAMs, indicating that induction of antiviral agents via TLR3 activation by Emodin might contribute to its anti-PRRSV effect. These findings imply that the Emodin from Aloe could hamper the proliferation of PRRSV in vitro and might constitute a new approach for treating PRRSV infection.

Correlation between serum 25-(OH)D3 level and immune imbalance of Th1/Th2 cytokines in patients with Hashimoto's thyroiditis and its effect on autophagy of human Hashimoto thyroid cells.

The study aimed to determine the relationship between serum 25-(OH)D3 and Th1/Th2 cytokine immune imbalance, and the effect of 25-(OH)D3 on the autophagy of human Hashimoto thyroid cells. Western blot analysis was used to detect the expression levels of microtubule-associated protein 1 light chain 3 (LC3) and autophagy-associated protein mammalian target protein of rapamycin (mTOR) in thyroid tissues of 20 Hashimoto's thyroiditis (HT) patients and normal tissues of 20 benign thyroid adenomas. Nthy-ori3-1 cells (normal cells of human thyroid follicular epithelium) were treated with different concentrations of 25-(OH)D3 for 24 h. The expression of LC3, mTOR and caspase-3 protein in the cells was detected by western blot analysis. The apoptosis and proliferation levels were detected by flow cytometry and MTT assay, respectively. The levels of FT3, FT4 and IL-10 in the HT group were lower than those in the healthy control group. The serum levels of 25-(OH)D3, TPOAb and TGAb in the HT group were lower than those in the healthy control group. Serum 25-(OH)D3 level in the HT group was negatively correlated with IL-2 and IFN-γ, and positively correlated with IL-4. In Hashimoto's thyroiditis tissues, the expression of mTOR was higher while the expression of LC3B-II was lower than that of normal thyroid tissue. With the increase in 25-(OH)D3 concentration, the expression level of mTOR increased, the expression level of LC3B-II decreased and the apoptosis rate was significantly increased. The cell proliferation rate decreased with the increase in 25-(OH)D3 concentration. The serum 25-(OH)D3 level in HT hypothyroidism patients was significantly lower than that of the control group. Thus, 25-(OH)D3 may be involved in the disease progression by upregulating the levels of Th1 cytokines and downregulating the levels of Th2 cytokines. 25-(OH)D3 can inhibit autophagy of thyroid cells, induce apoptosis and participate in the pathogenesis of Hashimoto's thyroiditis.

Epidemiological characteristics and associated factors of delayed voiding among middle school students / 中国学校卫生

Objective@#To analyze the epidemiological characteristics and associated factors of delayed voiding among middle school students, and to provide the basis for health education.@*Methods@#A total of 1 260 middle school students were selected from Sanmen country in Taizhou by using multi-stage stratified sampling method. A cross-sectional survey was conducted to investigate the demographic characteristics, epidemiological characteristics and associated factors of delayed voiding by questionnaire.@*Results@#The proportion of frequent delayed voiding among middle school students was 14.37%(181). The proportion among senior high school students was 15.51%(90), and among junior high school students was 13.67%(91)，no significant differences were found by grade(χ2=0.57, P=0.45). The multivariate Logistic regression analysis showed that residential school, homework per day >2 hours, games and reduce the frequency of drinking ≥7 times/week were risk factors for delayed voiding frequently in junior high school students(OR=3.71, 1.70, 3.02,6.52，P<0.05). Higher than 2 hours of daily homework, a supportive or indifferent attitude to delayed voiding, staying in bed after waking up, drinking <0.5 L/d or ≥2.0 L/d, and reduce the frequency of drinking ≥7 times/week were risk factors for delayed voiding behavior in high school students(OR=2.23, 1.81, 2.22, 3.48, 2.69, 3.00, P<0.05).@*Conclusion@#The prevalence of frequent delayed voiding was high among middle school students. Attitude, environment and the quality of drinking water were the primary factors of frequent delayed voiding.