Effective-components combination alleviates PM2.5-induced inflammation by evoking macrophage autophagy in COPD.

ETHNOPHARMACOLOGICAL RELEVANCE: Bufei Yishen formula (BYF) is clinically used to treat chronic obstructive pulmonary disease (COPD). Effective-component compatibility (ECC) is a combination of five active components derived from BYF, which has an equal effect on COPD to BYF. Our previous study has also demonstrated that ECC can protect COPD rats against PM2.5 exposure. However, the precise mechanisms remain to be elucidated. AIM OF THE STUDY: To explore the mechanism underlying the anti-inflammatory effects of ECC-BYF against PM2.5-accelerated COPD. MATERIALS AND METHODS: MH-S macrophages were stimulated by PM2.5 suspension to establish an in vitro model. Western blotting and immunofluorescent staining were used to measure the protein levels of autophagy markers. ELISA and quantitative PCR were used to detect the levels of inflammatory cytokines. In vivo, an established PM2.5-accelerated COPD rat model was used to determine the protective effect of ECC-BYF. Lung function, pathology, autophagy, and inflammatory mediators were detected. RESULTS: Firstly, we observed a significantly increased number of macrophages in the lungs upon PM2.5 exposure. Then, decreased autophagy flux while elevated inflammation was detected in PM2.5-exposed rats and MH-S cells. In MH-S cells, ECC-BYF significantly suppressed the PM2.5-increased inflammatory cytokines production, which was accompanied by the enhancement of autophagy flux. An autophagy inhibitor counteracted the anti-inflammatory effect elicited by ECC-BYF. In addition, ECC-BYF stimulated Foxo3 nuclear translocation and upregulated Foxo3 expression, whereas Foxo3 knockdown abrogated the inhibitory effect of ECC-BYF on inflammation. In PM2.5-accelerated COPD rats, ECC-BYF also attenuated the autophagy disruption and increased Foxo3 in the lungs, finally resulting in a suppression of pulmonary inflammation and an enhancement of lung function. CONCLUSION: ECC-BYF can ameliorate PM2.5-aggravated inflammation in COPD, which might be associated with the enhancement of autophagy flux in alveolar macrophages through the activation of Foxo3 signals.

Clinical Management of Low Anterior Resection Syndrome: Review of the Current Diagnosis and Treatment.

BACKGROUND: Low anterior resection syndrome (LARS) is a series of bowel dysfunction symptoms, including altered bowel frequency, irregular bowel rhythms, fecal incontinence, and constipation. LARS occurs in 80% of patients undergoing sphincter-preserving surgery, affecting patients' quality of life along with social avoidance. Different measurements and treatments have been raised to deal with LARS, but no systematic standard has been developed. OBJECTIVE AND METHODS: To promote the standardization of clinical trials and clinical management of LARS, this review summarizes the latest findings up until 2023 regarding the diagnostic criteria, assessment protocols, and treatment modalities for postoperative LARS in rectal cancer. RESULTS: The diagnostic criteria for LARS need to be updated to the definition proposed by the LARS International Collaborative Group, replacing the current application of the LARS score. In both clinical trials and clinical treatment, the severity of LARS should be assessed using at least one symptom assessment questionnaire, the LARS score or MSKCC BFI, and at least one scale related to quality of life. Anorectal manometry, fecoflowmetry, endoscopic ultrasonography, and pelvic floor muscle strength testing are recommended to be adopted only in clinical trials. After analysis of the latest literature on LARS treatment, a stepwise classification model is established for the standardized clinical management of LARS. Patients with minor LARS can start with first-line treatment, including management of self-behavior with an emphasis on diet modification and medication. Lamosetron, colesevelam hydrochloride, and loperamide are common antidiarrheal agents. Second-line management indicates multi-mode pelvic floor rehabilitation and transanal irrigation. Patients with major LARS should select single or several treatments in second-line management. Refractory LARS can choose antegrade enema, neuromodulation, or colostomy. CONCLUSIONS: In clinical trials of LARS treatment between 2020 and 2022, the eligibility criteria and evaluation system have been variable. Therefore, it is urgent to create a standard for the diagnosis, assessment, and treatment of LARS. Failure to set placebos and differentiate subgroups are limitations of many current LARS studies. Randomized controlled trials comparing diverse therapies and long-term outcomes are absent, as well. Moreover, a new scale needs to be developed to incorporate the patient's perspective and facilitate outpatient follow-up. Though the establishment of a stepwise classification model for LARS treatment here is indispensable, the refinement of the guidelines may be improved by more standardized studies.

First Report of Root Rot Caused by Ilyonectria robusta in the Medicinal Herb Aconitum carmichaelii in China.

Aconitum carmichaelii Debeaux is used as a traditional Chinese medicine with antiarrhythmic, antiinflammatory and other pharmacological functions. It is widely cultivated in China. According to our survey, about 60% of A. carmichaelii in Qingchuan, Sichuan, suffered from root rot, reducing yields by 30% in the past five years. Symptomatic plants exhibited stunted growth, dark brown roots, reduced root biomass, and fewer root hairs. The disease caused root rot and plant death in 50% of the infected plants. In October 2019, ten symptomatic 6-month-old plants were collected from fields in Qingchuan. Diseased pieces of the roots were surface sterilized with sodium hypochlorite solution (2%), rinsed three times in sterile water, plated on potato dextrose agar (PDA), and incubated at 25°C in the dark. Six single-spore isolates of a Cylindrocarpon-like anamorp were obtained. The colonies on PDA were 35 to 37 mm diam after seven days with regular margins. The plates were covered with felty aerial mycelium, white to buff, and the reverse side chestnut near center with a ochre to yellowish leading edge. On spezieller nährstoffarmer agar (SNA), macroconidia were 1 to 3 septate, straight or slightly curved, cylindrical, with rounded ends, and varied in size: 1-septate 15.1 to 33.5 × 3.7 to 7.3 µm (n=250), 2-septate 16.5 to 48.5 × 3.7 to 7.6 µm (n=85), and 3-septate 22.0 to 50.6 × 4.9 to 7.4 µm (n=115). Microconidia were ellipsoid to ovoid, and 0 to 1 septate; aseptate spores were 4.5 to 16.8 × 1.6 to 4.9 µm (n=200), and 1-septate spores were 7.4 to 20.0 × 2.4 to 5.1 µm (n=200). The chlamydospores were brown, thick-walled, globose to subglobose, 7.9 to 15.9 µm (n=50). The morphology of these isolates was consistent with the previous description of Ilyonectria robusta (Cabral et al. 2012). Isolate QW1901 was characterized by sequencing the ITS, TUB, H3, and tef1α loci using previously reported primer pairs: ITS1/ITS4 (White et al. 1990), T1/Bt-2b (O'Donnell and Cigelnik 1997), CYLH3F/CYLH3R (Crous et al. 2004), and EF1/EF2 (O'Donnell et al. 1998). A Blastn search of the sequences of ITS, TUB, H3, and tef1α showed that QW1901 shared 99.26, 97.89, 97.79, and 99.17 % identities, respectively, with the ex-type strain of I. robusta (CBS308.35). The ITS, TUB, H3, and tef1α sequences were deposited in GenBank under accession nos. MW534715, and MW880180 to MW880182, respectively. A phylogenetic tree was constructed from a neighbor-joining analysis on the alignment of the combined ITS, TUB, H3, and tef1α sequence. QW1901 was clustered with the ex-type strain of I. robusta. To confirm the pathogenicity of I. robusta, bare roots of healthy 6-month-old A. carmichaelii were inoculated with mycelial plugs of 7-day-old QW1901 colonies selected randomly (Lu et al. 2015). Five needle-wound lateral roots and five intact roots were inoculated as replicates with pathogen-free agar plugs as a control. Then, all plants were grown in sterile soil in a growth chamber at 20±1°C and watered regularly. Pathogenicity assays were repeated twice. After 20 days of cultivation, infected plants exhibited symptoms similar to those observed in the field. All control plants remained asymptomatic. Sequencing confirmed the re-isolation of I. robusta from the inoculated plants, satisfying Koch's hypothesis. Ilyonectria robusta has been reported to cause root rot of plants such as Codonopsis tangshen and Panax ginseng ( Lu et al. 2015; Zheng et al. 2021), and has also been reported to be isolated from Aconitum kongboense in China (Wang et al. 2015). However, this is the first report of the pathogen causing root rot of A. carmichaelii. Management measures, such as growing disease-free seedlings in sterile soil, should be used to minimize the risk of this pathogen.

A comparison of exosomes derived from different periods breast milk on protecting against intestinal organoid injury.

AIM OF THE STUDY: Human breast milk reduces the risk and severity of necrotizing enterocolitis (NEC). Exosomes are extracellular vesicles (EVs) found in high concentrations in milk, and they mediate intercellular communication and immune responses. The aim of this study is to compare the protective effects of exosomes that are derived from different time periods of breast milk production against intestinal injury using an ex vivo intestinal organoid model. METHODS: Colostrum, transitional and mature breast milk samples from healthy lactating mothers were collected. Exosomes were isolated using serial ultracentrifugation and filtration. Exosomes' presence was confirmed using transmission electron microscopy (TEM) and western blot. To form the intestinal organoids, terminal ileum was harvested from neonatal mice pups at postnatal day 9, crypts were isolated and organoids were cultured in matrigel. Organoids were either cultured with exposure to lipopolysaccharide (LPS), or in treatment groups where both LPS and exosomes were added in the culturing medium. Inflammatory markers and organoids viability were evaluated. MAIN RESULTS: Human milk-derived exosomes were successfully isolated and characterized. LPS administration reduced the size of intestinal organoids, induced inflammation through increasing TNFα and TLR4 expression, and stimulated intestinal regeneration. Colostrum, transitional and mature human milk-derived exosome treatment all prevented inflammatory injury, while exosomes derived from colostrum were most effective at reducing inflammatory cytokine. CONCLUSIONS: Human breast milk-derived exosomes were able to protect intestine organoids against epithelial injury induced by LPS. Colostrum exosomes offer the best protective effect among the breast-milk derived exosomes. Human milk exosomes can be protective against the development of intestinal injury such as that seen in NEC.

Cockroach sensitization mitigates allergic rhinoconjunctivitis symptom severity in patients allergic to house dust mites and pollen.

BACKGROUND: Modifiers of symptom severity in patients with allergic rhinoconjunctivitis (AR) are imprecisely characterized. The hygiene hypothesis implicates childhood microbial exposure as a protective factor. Cockroach sensitization (C+) might be a proxy for microbial exposure. OBJECTIVE: We sought to determine whether C+ assayed by means of skin prick tests influenced AR symptom severity in controlled and natural settings. METHODS: Total symptom scores (TSSs) were recorded by 21 participants with house dust mite allergy (M+) in the natural setting and during repeated exposures of 3 hours per day to house dust mite allergen in an allergen challenge chamber (ACC). In M+ participants the peripheral blood and nasal cells were assayed for T-cell activation and transcriptomic profiles (by using RNA sequencing), respectively. Participants allergic to mountain cedar (n = 21), oak (n = 34), and ragweed (n = 23) recorded TSSs during separate out-of-season exposures to these pollens (any pollen sensitization [P+]) in the ACC; a subset recorded TSSs in the pollination seasons. RESULTS: The hierarchy of TSSs (highest to lowest) among M+ participants tracked the following skin prick test sensitization statuses: M+P+C- > M+P+C+ > M+P-C- > M+P-C+. In nasal cells and peripheral blood the immune/inflammatory responses were rapidly resolved in M+P+C+ compared with M+P+C- participants. Among those allergic to pollen, C+ was associated with a lower TSS during pollen challenges and the pollination season. After aggregated analysis of all 4 ACC studies, C+ status was associated with a 2.8-fold greater likelihood of a lower TSS compared with C- status (odds ratio, 2.78; 95% CI, 1.18-6.67; P = .02). CONCLUSIONS: C+ status is associated with mitigation of AR symptom severity in adults with AR.

Effect of confounding cofactors on responses to pollens during natural season versus pollen challenge chamber exposure.

BACKGROUND: The severity of allergic rhinoconjunctivitis (AR) symptomatology elicited after exposure to pollen in the absence versus the presence of confounding cofactors, such as in a pollen challenge chamber (PCC) and the natural pollinating season, respectively, might differ. OBJECTIVE: We sought to determine the correlation of AR severity in the natural season versus out-of-season PCC exposures. METHODS: Twenty-four Virginia live oak (VLO)-positive, 14 VLO-negative, 16 mountain cedar (MC)-positive, 8 MC-negative, and 26 ragweed-positive participants recorded AR symptoms (total symptom score [TSS]) during the VLO, MC, and ragweed pollinating seasons and during 2 consecutive PCC exposures of 3 hours each to these pollens separately. RESULTS: The TSSs recorded before the natural season were higher than the pre-PCC values. This prepriming was greater among VLO(+) than MC(+) participants, and it blunted further increases in TSSs during the VLO natural season. Nonatopic participants were nonreactive in the PCC. There was wide variation in the level of AR symptomatology after exposure to VLO, MC, or ragweed pollen in the PCC. Prepriming formed the basis for higher AR responses observed in the natural season than in the PCC, resulting in the identification of distinct PCC/natural season endophenotypes and a partial correlation between the TSSs recorded in the natural season versus those recorded in the PCC (r = 0.34, 0.54, and 0.65 for VLO(+), MC(+), and ragweed-positive participants, respectively). CONCLUSIONS: Prepriming in the natural pollinating season might obscure the true correlation between AR severity in the natural season versus the PCC. By mitigating confounding cofactors, PCC exposures have utility for evaluation of novel AR therapeutics.

Responses to ragweed pollen in a pollen challenge chamber versus seasonal exposure identify allergic rhinoconjunctivitis endotypes.

BACKGROUND: The level of concordance between allergic symptoms induced on exposure to pollen in a pollen challenge chamber (PCC) versus the natural season is unknown. OBJECTIVE: We sought to test the hypothesis that the symptom levels of allergic rhinoconjunctivitis elicited after out-of-season exposure to short ragweed in a PCC and during the natural season for giant ragweed pollen are highly correlated. METHODS: Thirty-one ragweed-sensitive participants recorded symptoms for 15 days during the natural giant ragweed season in San Antonio, Texas. Twenty-six of these participants were challenged to short ragweed pollen in a PCC for 3 hours per day for up to 4 days. RESULTS: In the PCC participants were dichotomized into those in whom low versus high levels of symptoms developed slowly or rapidly (ie, slow/low vs rapid/high). Each successive exposure visit associated with a progressive increase in symptom levels that approximated those experienced during the natural season. Hierarchic clustering identified 3 endotypes: endotypes I and II reflected concordantly low (n= 7) versus high (n = 14) total symptom scores (TSSs) in both the natural season and the PCC, respectively. Accordingly, the correlation between the TSSs recorded in the natural season and in the PCC for these 21 participants was very high. Although participants with endotype III (n = 5) had greater TSSs in the natural season than in the PCC, the degree of correlation between the TSSs remained high. CONCLUSIONS: Our findings affirm our hypothesis, underscore the high cross-reactivity between distinct pollens, and highlight the utility of the PCC to identify novel allergy endotypes that might have contrasting mechanistic underpinnings and potentially therapeutic responses.