Apolipoprotein L3 inhibits breast cancer proliferation and modulates cell cycle via the P53 pathway.

Background: Breast cancer is the second most common cause of cancer-related mortality globally. Apolipoprotein L3 (APOL3), a member of the apolipoprotein family, has been implicated in the pathogenesis of cardiovascular diseases. Nevertheless, the functions and underlying mechanisms of APOL3 in breast cancer have yet to be elucidated. Methods: The patient data were sourced from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database. Quantitative real-time PCR (qRT-PCR), western blotting, and immunohistochemistry (IHC) assays were used to assess expression of APOL3. Cell proliferation rates were determined by Cell Counting Kit-8 (CCK-8) and colony formation assays. Flow cytometry was used to examine cell cycle distribution. Western blotting was conducted to investigate the expression of cell cycle related proteins. A xenograft model was used to evaluate the effect of APOL3 in vivo. APOL3-binding proteins were identified through mass spectrometry, co-immunoprecipitation (CO-IP) assay and immunofluorescence assay. Results: APOL3 expression was significantly downregulated in breast cancer, and its low expression was correlated with poor prognostic outcomes. Overexpression of APOL3 suppressed breast cancer cell proliferation, induced cell cycle disruption. Conversely, knockdown of APOL3 promoted cell proliferation. In vivo animal experiments demonstrated that APOL3 overexpression can inhibit tumor proliferation. Mass spectrometry, CO-IP and immunofluorescence assay confirmed the interaction between APOL3 and Y-box binding protein 1 (YBX1). Furthermore, YBX1 knockdown following APOL3 knockdown mitigated the enhanced proliferation. These results provide new ideas for clinically targeting APOL3 to inhibit proliferation in breast cancer. Conclusions: Our findings indicate that APOL3 inhibits breast cancer cell proliferation and cell cycle modulating P53 pathway through the interaction of YBX1.

Digital Immunoassay for Rapid Detection of SARS-CoV-2 Infection in a Broad Spectrum of Animals.

The ability of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) to infect a wide-range of species raises significant concerns regarding both human-to-animal and animal-to-human transmission. There is an increasing demand for highly sensitive, rapid, and simple diagnostic assays that can detect viral infection across various species. In this study, we developed a biosensor assay that adapted a monoclonal-antibody (mAb)-based blocking ELISA format into an Activate Capture + Digital Counting (AC + DC)-based immunoassay. The assay employs a photonic crystal (PC) biosensor, gold-nanoparticle (AuNP) tags, SARS-CoV-2 nucleocapsid (N) protein, and specific anti-N mAb to detect antibody responses in animals exposed with SARS-CoV-2. We demonstrated a simple 2-step 15-min test that was capable of detecting as low as 12.5 ng of antibody in controlled standard serum samples. Based on an evaluation of 176 cat serum samples with known antibody status, an optimal percentage of inhibition (PI) cut-off value of 0.588 resulted in a diagnostic sensitivity of 98.3% and a diagnostic specificity of 96.5%. The test is highly repeatable with low variation coefficients of 2.04%, 2.73%, and 4.87% across different runs, within a single run, and on a single chip, respectively. The test was further employed to detect antibody responses in multiple animal species as well as investigate dynamics of antibody response in experimentally infected cats. This test platform provides an important tool for rapid field surveillance of SARS-CoV-2 infection across multiple species.

Successful treatment with tislelizumab plus chemotherapy for SMARCA4-deficient undifferentiated tumor: a case report.

SMARCA4-deficient undifferentiated tumor (SMARCA4-dUT) is a devastating subtype of thoracic tumor with SMARCA4 inactivation and is characterized by rapid progression, poor prognosis, and high risk of postoperative recurrence. However, effective treatments for SMARCA4-dUT are lacking. Herein, we describe a patient with SMARCA4-dUT who exhibited an impressive response to the anti-programmed cell death protein-1 (PD-1) antibody (tislelizumab) in combination with conventional chemotherapy (etoposide and cisplatin). To the best of our knowledge, this is the first case of SMARCA4-dUT treated with chemotherapy, comprising etoposide and cisplatin, combined with anti-PD-1 inhibitors. Immunotherapy combined with etoposide and cisplatin may be a promising strategy to treat SMARCA4-dUT.

Osteoporosis: Emerging targets on the classical signaling pathways of bone formation.

Osteoporosis is a multifaceted skeletal disorder characterized by reduced bone mass and structural deterioration, posing a significant public health challenge, particularly in the elderly population. Treatment strategies for osteoporosis primarily focus on inhibiting bone resorption and promoting bone formation. However, the effectiveness and limitations of current therapeutic approaches underscore the need for innovative methods. This review explores emerging molecular targets within crucial signaling pathways, including wingless/integrated (WNT), bone morphogenetic protein (BMP), hedgehog (HH), and Notch signaling pathway, to understand their roles in osteogenesis regulation. The identification of crosstalk targets between these pathways further enhances our comprehension of the intricate bone metabolism cycle. In summary, unraveling the molecular complexity of osteoporosis provides insights into potential therapeutic targets beyond conventional methods, offering a promising avenue for the development of new anabolic drugs.

BMP2-ERK-ATF4 Axis-Based 6-methoxybenzofuran Compound I-9 Acts as Candidate Drug for Bone Formation and Anti-Osteoporosis.

As the global population ages, the number of patients with osteoporosis is rapidly rising. The existing first-line clinical drugs are bone resorption inhibitors that have difficulty restoring the bone mass of elderly patients to the safe range. The range and period of use of existing peptides and monoclonal antibodies are limited, and small-molecule bone formation-promoting drugs are urgently required. We established an I-9 synthesis route with high yield, simple operation, and low cost that was suitable for future large-scale production. I-9 administration promoted bone formation and increased bone mass in mice with low bone mass in an aged C57 mouse model. Our findings revealed a hitherto undescribed pathway involving the BMP2-ERK-ATF4 axis that promotes osteoblast differentiation; I-9 has favorable biosafety in mice. This study systematically investigated the efficacy, safety, and mechanism of I-9 for treating osteoporosis and positions this drug for preclinical research in the future. Thus, this study has promoted the development of small-molecule bone-promoting drugs.

Integration of network pharmacology with experimental validation to reveal the mechanism of action of Longdan Xiegan Decoction against HSV2 infection and determine its effective components.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese Medicine (TCM) has made enormous strides recently in the discovery of anti-herpes simplex virus (HSV) drugs under the guidance of TCM theory. Longdan Xiegan Decoction (LXD), a formulation recorded in the Pharmacopoeia of the People's Republic of China, has proved to be effective against HSV infection. However, its effective components and action mechanism remain unclear. AIM OF THE STUDY: To investigate the effective components and mechanisms of LXD in treating HSV infection based on network pharmacology and experimental validation. MATERIALS AND METHODS: The anti-HSV activities of key compounds predicted by network analysis were detected by antiviral tests. High-performance liquid chromatography (HPLC) was applied to identify the main components of the LXD aqueous extract. Time-of-addition assay and infectivity inhibition reversibility assay were conducted to identify the potential antiviral mechanisms of licochalcone B (LCB). Additionally, we assessed the antiviral effect of LCB in vivo by use of body weight, viral load, histological analysis, and scoring of genital lesions in an HSV2-infected mouse model. RESULTS: Our data demonstrated that some components exhibited significant anti-HSV1/2 activity in vitro, including quercetin, kaempferol, wogonin, formononetin, naringenin, baicalein, isorhamnetin, glabridin, licochalcone A, echinatin, oroxylin A, isoliquiritigenin, pinocembrin, LCB and acacetin. HPLC analysis showed that LCB was the main component of LXD aqueous extract. In vitro experiments revealed that LCB not only inactivated HSV2 particles, but also inhibited HSV2 multiplication through the inhibition of the phosphorylation of Akt and its downstream targets. In vivo experiments confirmed that LCB could significantly reduce viral titer, delay weight loss, and alleviate pathological changes in vaginal tissue in vaginal infection mouse models. CONCLUSION: LCB acted as the main component of LXD, with significant anti-HSV2 infection effects both in vivo and in vitro. This study provides additional evidence of the healing efficacy of LXD against HSV infection and presents an efficient analytical method for further investigation of the mechanisms of TCM in prevention and treatment of various diseases.

Comparison of the effect of capsular bend on the rotational stability between 2 toric intraocular lenses.

PURPOSE: To compare the effect of capsular bend on the rotational stability between 2 toric intraocular lenses (IOLs). SETTING: Eye Hospital of Wenzhou Medical University, Hangzhou, Zhejiang Province, China. DESIGN: Prospective study. METHODS: Patients with preexisting astigmatism received AcrySof IQ (SN6AT) or TECNIS (ZCT/ZMT) toric IOL during cataract surgery. CASIA2 was used to record the toric IOL axial orientation and capsular bend index (CBI) at the 1-day, 1-week, 1-month, and 3-month interval postoperatively. The postoperative rotational stability and CBI of both models were compared. RESULTS: A total of 58 eyes from 58 patients were enrolled in this study. The total misalignment of the TECNIS (ZCT/ZMT) group (6.96 ± 5.10 degrees, 7.41 ± 5.19 degrees, 6.93 ± 5.29 degrees, and 6.86 ± 5. 27 degrees) was significantly higher than that of the AcrySof IQ (SN6AT) group (3.55 ± 2.21 degrees, 4.00 ± 2.74 degrees, 3.72 ± 2.72 degrees, and 3.52 ± 2.50 degrees) at all follow-up intervals ( P < .05). The mean rotation of the TECNIS (ZCT/ZMT) group (2.66 ± 2.18 degrees) was significantly greater than that of the AcrySof IQ (SN6AT) group (1.65 ± 1.47 degrees) from 1 day to 1 week postoperatively ( P < .05). The capsular bend formation in the TECNIS (ZCT/ZMT) group was delayed compared with the AcrySof IQ (SN6AT) group ( P < .05, at the 1-week, 1-month, and 3-month interval). The TECNIS (ZCT/ZMT) group showed fibrosis in the peripheral anterior capsule, leading to its stretching away from the IOL surface, while the AcrySof IQ (SN6AT) group exhibited gentle adherence of the anterior capsule to the IOL surface. CONCLUSIONS: The AcrySof IQ toric IOL (SN6AT) exhibited greater rotational stability than the TECNIS toric IOL (ZCT/ZMT), which may partially result from the delay in capsular bend formation of TECNIS at the 1-day to 1-week follow-up postoperatively.

Design and synthesis of dabigatran etexilate derivatives with inhibiting thrombin activity for hepatocellular carcinoma treatment.

Hepatocellular carcinoma (HCC) is one of the most fatal solid malignancies worldwide. Evidence suggests that thrombin stimulates tumor progression via fibrin formation and platelet activation. Meanwhile, we also found a correlation between thrombin and HCC through bioinformatics analysis. Dabigatran is a selective, direct thrombin inhibitor that reversibly binds to thrombin. Dabigatran was used as the lead agent in this study, and 19 dabigatran derivatives were designed and synthesized based on docking mode. The thrombin-inhibitory activity of the derivative AX-2 was slightly better than that of dabigatran. BX-2, a prodrug of AX-2, showed a fairly strong inhibitory effect on thrombin-induced platelet aggregation, and effectively antagonized proliferation of HCC tumor cells induced by thrombin at the cellular level. Furthermore, BX-2 reduced tumor volume, weight, lung metastasis, and secondary tumor occurrence in nude mouse models. BX-2 combined with sorafenib increased sorafenib efficacy. This study lays the foundation for discovering new anti-HCC mechanism based on thrombin. BX-2 can be used as an anti-HCC drug lead for further research.

Unsupervised Learning Based on Meibography Enables Subtyping of Dry Eye Disease and Reveals Ocular Surface Features.

Purpose: This study aimed to establish an image-based classification that can reveal the clinical characteristics of patients with dry eye using unsupervised learning methods. Methods: In this study, we analyzed 82,236 meibography images from 20,559 subjects. Using the SimCLR neural network, the images were categorized. Data for each patient were averaged and subjected to mini-batch k-means clustering, and validated through consensus clustering. Statistical metrics determined optimal category numbers. Using a UNet model, images were segmented to identify meibomian gland (MG) areas. Clinical features were assessed, including tear breakup time (BUT), tear meniscus height (TMH), and gland atrophy. A thorough ocular surface evaluation was conducted on 280 cooperative patients. Results: SimCLR neural network achieved clustering patients with dry eye into six image-based subtypes. Patients in different subtypes harbored significantly different noninvasive BUT, significantly correlated with TMH. Subtypes 1 and 5 had the most severe MG atrophy. Subtype 2 had the highest corneal fluorescent staining (CFS). Subtype 4 had the lowest TMH, whereas subtype 5 had the highest. Subtypes 3 and 6 had the largest MG areas, and the upper MG areas of a person's bilateral eyes were highly correlated. Image-based subtypes are related to meibum quality, CFS, and morphological characteristics of MG. Conclusions: In this study, we developed an unsupervised neural network model to cluster patients with dry eye into image-based subtypes using meibography images. We annotated these subtypes with functional and morphological clinical characteristics.

UVEAL EFFUSION ASSOCIATED WITH LOCALIZED SCLERODERMA BECAUSE OF SCLERAL FIBROSIS.

BACKGROUND/PURPOSE: To report a case of uveal effusion associated with localized scleroderma because of scleral collagen fibrosis. Partial-thickness sclerectomy treatment was successful in acquiring the resolution of the uveal effusion. METHODS: Case report. RESULTS: A 44-year-old Chinese woman with known localized scleroderma visited the retinal clinic complaining of insidious onset blurring of vision in both eyes for 8 months. The best-corrected visual acuity was 20/200. Ophthalmoscopy revealed apparent inferior bullous serous retinal detachments in the right eye. Optical coherence tomography showed subretinal fluid and folds of the retinal pigment epithelium layer in both eyes. B-scan ultrasonographic image of the right eye confirmed a 360-degree serous retinal detachment in the right eye accompanied with increased thickness of the ocular wall. Ultrasound biomicroscopy of the anterior segment detected a shallow ciliary body detachment in the right eye. Fluorescein angiography and indocyanine green angiography demonstrated the leopard-spot pattern in all phases. Partial-thickness sclerectomy treatment was successful in acquiring the resolution of the uveal effusion. Histopathologic examinations of the sclera flaps revealed scleral collagen fibrosis. CONCLUSION: This clinicopathologic report first describes a patient with localized scleroderma and scleral collagen fibrosis, resulting in uveal effusion that responded to partial-thickness sclerectomy.

Treatment of visual axis opacification and secondary membranes with Nd:YAG laser after pediatric cataract surgery under intranasal sedation.

Purpose: To describe neodymium-doped yttrium-aluminum-garnet (Nd:YAG) laser treatment of visual axis opacification and secondary membranes in pediatric patients with cataracts under intranasal dexmedetomidine sedation. Methods: Twenty eyes of 17 patients with secondary membrane formation after cataract extraction were enrolled in this study. Intranasal dexmedetomidine sedation (3 ug/kg) was administered, and Nd:YAG laser (Ellex Super Q, Adelaide, Australia) procedures were performed with children in the sitting position with their chin supported on a laser delivery slit lamp. Preoperative and postoperative visual acuities were documented, and medical records were reviewed. Results: The age of the patients ranged from 5 to 83 months (31.82 ± 27.73). Nineteen (95.0%) eyes had congenital cataracts and one (5.0%) had a traumatic cataract. Nd:YAG laser treatment of VAO with ten (50.0%) eyes, pupillary membranes with three (15.0%) eyes, pupillary cortical proliferation with six (30.0%) eyes, and anterior capsule contraction with one (5.0%) eye. Five (25.0%) eyes demonstrated visual acuity improvement, whereas six (30.0%) eyes remained unchanged after laser treatment. The recurrence rate was 30.0% and four eyes underwent a second Nd:YAG membranectomy. No side effects or tolerances due to sedative drugs were observed. Conclusion: Nd:YAG laser membranectomy under intranasal dexmedetomidine sedation was safely performed in children as young as 5 months old in a sitting position. This approach facilitates patient convenience, doctor proficiency, and cost reductions. Patients with recurrence can be treated by repeating the procedure.

Femtosecond Laser-Assisted Cataract Surgery versus Conventional Phacoemulsification Surgery: Clinical Outcomes with EDOF IOLs.

In this study, we evaluate and compare the outcomes of conventional phacoemulsification cataract surgery (CPS) and femtosecond laser-assisted cataract surgery (FLACS) with the implantation of an extended depth of field (EDOF) intraocular lens (IOL). A prospective, consecutive cohort study was conducted. Patients were given the option to choose FLACS or CPS and were implanted with an EDOF IOL. Refractive data, visual acuity data, ocular aberration measured with a wavefront aberrometer, and optical quality measured with an optical quality analysis system II were collected at one month postoperatively. A total of 92 eyes of 64 patients were enrolled in this study; 35 eyes of 26 patients were treated with FLACS, whereas 57 eyes of 38 patients were treated with CPS. Uncorrected visual acuity at far, intermediate, and near distance and best-spectacle-corrected visual acuity were not statistically significantly different between the groups (all p > 0.05), nor were the mean cylinder and mean spherical equivalent refraction (both p > 0.05). The FLACS group had a lower ocular trefoil than the CPS group (p = 0.033), and there was no significant difference between the two groups considering other aberration parameters, whether ocular or internal (all p > 0.05). Optical-quality-related parameters showed also no significant difference between the two groups (all p > 0.05). In conclusion, there was no significant difference between FLACS and CPS with implantation of EDOF IOLs in postoperative ocular parameters, refractive outcomes, ocular aberration, optical quality, and aberration parameters, except a lower ocular trefoil in the FLACS group. In terms of these indicators, FLACS does not provide an additional clinical benefit for patients over CPS.

Paeonol inhibits melanoma growth by targeting PD1 through upregulation of miR-139-5p.

The abnormal immune response mediated by malignant melanoma is related to PD1. Paeonol has pharmacological antitumor activity. Previous studies have indicated that paeonol induces tumor cell apoptosis, but its underlying mechanism in tumor immunity remains unknown. In this study, malignant melanoma was established in normal and thymectomized mice to determine the important role of the thymus in the antitumor effects of paeonol. Paeonol-treated thymocytes were cocultured with melanoma cell spheres to further evaluate the regulatory role of thymocytes in tumor immune dysfunction. Studies have shown that PD1 may be targeted by miR-139-5p. Our results revealed that tumor-induced thymic atrophy was significantly accompanied by high PD1 expression and low miR-139-5p expression. Interestingly, paeonol significantly reversed thymic atrophy and largely protected thymocytes against low PD1 expression and high miR-139-5p expression. Dual-luciferase assays indicated that miR-139-5p interacted with the 3' untranslated region (3'-UTR) of PD1. These results showed that paeonol alleviates PD1-mediated antitumor immunity by reducing miR-139-5p expression and demonstrated a novel mechanism for melanoma immunotherapy.

Do Descriptive Norm Appeals in Public Service Ads Reduce Freedom Threats? Examining the Effects of Normative Messages and Media Literacy Skills on Decreasing Reactance.

While health communication campaigns seek to encourage and promote healthy behaviors, they are not always successful. Health communication efforts may fail for several reasons, such as viewers experiencing excessive freedom threats and reactance. This study (n = 201) proposes and demonstrates that descriptive norm appeals in health PSAs can indirectly lead to enhanced behavioral intentions toward the message advocacy via reducing perceived freedom threats, inhibiting psychological reactance, and improving message credibility. However, this serial mediation was only found for message viewers with relatively low media literacy skills - precisely, who did not show considerable critical thinking toward media content. For participants who reported a high level of critical thinking toward media content, the use of descriptive norm appeals did not decrease freedom threats, nor did it indirectly affect behavioral intentions. The findings of this study contribute to the theory of psychological reactance and norms-based research. Both theoretical and practical implications are provided for health communication scholars and practitioners.

Accuracy of new-generation intraocular lens calculation formulas in eyes with variations in predicted refraction.

PURPOSE: To investigate the characteristics of eyes with large variations in predicted refraction using four traditional intraocular lens (IOL) formulas and evaluate the accuracy of new-generation intraocular lens power calculation formulas. METHODS: Eyes that had variation in predicted refraction (≥ 0.75 D) using four traditional formulas (SRK/T, Holladay 1, Hoffer Q, and Haigis formulas) were included. Axial length (AL), anterior chamber depth (ACD), average keratometry (AK), and the ratio of axial length to corneal radius (AL/CR) were measured. New-generation formulas (Barrett Universal II, Emmetropia Verifying Optical 2.0, Kane, and Pearl-DGS formulas) and traditional formulas were compared. The median absolute error (MedAE) was the main parameter to evaluate the accuracy of formulas. RESULTS: A total of 79 participants (79 eyes) who had variation in predicted refraction of (≥ 0.75 D) using four traditional formulas out of 510 eyes (510 patients) underwent uncomplicated cataract surgeries. The Barrett Universal II (0.29 D), EVO 2.0 (0.31 D), Kane (0.30 D), and Pearl-DGS (0.33 D) formulas produced significantly lower median absolute errors (MedAEs) than the Hoffer Q (0.61 D) and Holladay 1 (0.59 D) formulas (P < 0.01). The Wang-Koch (WK) adjustment significantly improved the accuracy of the Holladay 1 formula in long eyes (P < 0.001). CONCLUSIONS: Abnormal AL, ACD, and AK are more likely to lead to prediction errors using traditional formulas. New-generation formulas and traditional formulas with WK adjustment showed satisfactory prediction accuracy.

Breast cancer gene expression signatures: development and clinical significance-a narrative review.

Background and Objective: Breast cancer gene expression signatures are developing rapidly and are expected to better understand the intrinsic features of the tumor, and also to optimize the treatment strategy in clinical practice. This review is to summarize the controversy and consensus in clinical practice of gene expression signatures, and to provide our perspective on these issues as well as recommendation for future direction. Methods: We reviewed English publications in PubMed related to breast cancer gene expression signatures from 2002 to 2022. Key Content and Findings: Five mature commercial gene expression signatures: Oncotype, MammaPrint, Prosigna/PAM50, EndoPredict and Breast Cancer Index (BCI) are available to provide the prognostic and predictive assessment. Although they could help to evaluate the risk of recurrence and to predict the benefits of certain treatments, their applications remain challenging. Treatment decisions should be determined by a combination of related clinical pathological factors in clinical practice. Conclusions: Gene expression signatures could assist in the determination of the adjuvant therapy of early-stage breast cancer. The prospective randomized clinical trials showed that chemotherapy may be exempted in low-risk patients. More sufficient data are expected for the application in radiotherapy, extended endocrine therapy, and neoadjuvant treatment. The treatment cannot be determined by a single factor but by comprehensive assessments of clinicopathological factors, test purpose, and cost-effectiveness. Patients will benefit from personalized treatments with the publication of further evidence.

Structure-based virtual screening identified novel FOXM1 inhibitors as the lead compounds for ovarian cancer.

Ovarian cancer (OC) is a gynecological tumor with possibly the worst prognosis, its 5-year survival rate being only 47.4%. The first line of therapy prescribed is chemotherapy consisting of platinum and paclitaxel. The primary reason for treatment failure is drug resistance. FOXM1 protein has been found to be closely associated with drug resistance, and inhibition of FOXM1 expression sensitizes cisplatin-resistant ovarian cancer cells. Combining existing first-line chemotherapy drugs with FOXM1 prolongs the overall survival of patients, therefore, FOXM1 is considered a potential therapeutic target in ovarian cancer. Previous research conducted by our team revealed a highly credible conformation of FOXM1 which enables binding by small molecules. Based on this conformation, the current study conducted virtual screening to determine a new structural skeleton for FOXM1 inhibitors which would enhance their medicinal properties. DZY-4 showed the highest affinity towards FOXM1, and its inhibitory effect on proliferation and migration of ovarian cancer at the cellular level was better than or equal to that of cisplatin, while its efficacy was equivalent to that of cisplatin in a nude mouse model. In this study, the anti-tumor effect of DZY-4 is reported for the first time. DZY-4 shows potential as a drug that can be used for ovarian cancer treatment, as well as a drug lead for future research.

Talking About Prescription Opioid Misuse: The Effect of Family Communication Patterns and Messages Advocating Direct and Indirect Communication.

Families play an important role in addressing substance misuse and addiction. Extant literature suggests patterns of communication within families influence the ways in which they engage loved ones who may be misusing substances like prescription opioids. However, little is known regarding how strategic health messages about family communication influence individuals' intentions to engage in conversations about substance misuse. Applying a normative approach, we conducted a (2 × 2) between-participants experiment examining whether messages advocating indirect (versus direct) communication are more effective for individuals (n = 613) who describe their family as having a low (versus high) conversation orientation. Univariate analysis of variance tests show match effects for message attitudes and message elaboration. For intentions to talk with a loved one about the risks of OUD, there was only evidence of a matching effect between the message advocating indirect communication with low conversation audiences. Both message types were equally effective at influencing intentions for high conversation participants. These findings suggest message designers should consider the kinds of communication behaviors and actions advocated in appeals targeting family members. Messages that are inclusive of the conversation dynamics of particular audiences may have greater effect. In particular, for low conversation audiences, messages advocating an indirect approach may be more effective at motivating intentions to engage someone who is misusing opioids.

Identification of phytochemicals in Qingfei Paidu decoction for the treatment of coronavirus disease 2019 by targeting the virus-host interactome.

Qingfei Paidu decoction (QFPDD) has been clinically proven to be effective in the treatment of coronavirus disease 2019 (COVID-19). However, the bioactive components and therapeutic mechanisms remain unclear. This study aimed to explore the effective components and underlying mechanisms of QFPDD in the treatment of COVID-19 by targeting the virus-host interactome and verifying the antiviral activities of its active components in vitro. Key active components and targets were identified by analysing the topological features of a compound-target-pathway-disease regulatory network of QFPDD for the treatment of COVID-19. The antiviral activity of the active components was determined by a live virus infection assay, and possible mechanisms were analysed by pseudotyped virus infection and molecular docking assays. The inhibitory effects of the components tested on the virus-induced release of IL-6, IL-1ß and CXCL-10 were detected by ELISA. Three components of QFPDD, oroxylin A, hesperetin and scutellarin, exhibited potent antiviral activities against live SARS-CoV-2 virus and HCoV-OC43 virus with IC50 values ranging from 18.68 to 63.27 µM. Oroxylin A inhibited the entry of SARS-CoV-2 pseudovirus into target cells and inhibited SARS-CoV-2 S protein-mediated cell-cell fusion by binding with the ACE2 receptor. The active components of QFPDD obviously inhibited the IL-6, IL-1ß and CXCL-10 release induced by the SARS-CoV-2 S protein. This study supports the clinical application of QFPDD and provides an effective analysis method for the in-depth study of the mechanisms of traditional Chinese medicine (TCM) in the prevention and treatment of COVID-19.