Association between life-ever gallstones and depressive symptoms in U.S. adults: a cross-sectional study.

Research on the potential association between life-ever gallstones and depressive symptoms is limited. This study aims to evaluate whether the presence of gallstone disease is associated with depressive symptoms. In this cross-sectional study, we analyzed data from the National Health and Nutrition Examination Survey (NHANES) 2017-March 2020 cycles. The presence of depressive symptoms and gallstone disease was assessed using questionnaire responses. Adjusted odds ratios (OR) were calculated using a multivariate logistic regression model, with adjustments made for age, sex, race, body mass index, history of cardiovascular disease, hypertension, arthritis, and pulmonary disease across different models. Subgroup and sensitivity analyses were conducted to ensure the stability of the results. This study included 6201 adults aged 20 years and above, with 539(8.7%) experiencing depressive symptoms. After adjusting for age, sex, race, body mass index, CVD history, hypertension, arthritis, pulmonary disease, depressive symptoms were possibly associated with life-ever gallstones (OR 1.37, 95% CI 0.91-2.08).When depressive symptoms were categorized as mild, moderate, moderately severe, and severe,life-ever gallstones was possibly associated with mild depressive symptoms (OR 1.12, 95% CI 0.81-1.56), moderate depressive symptoms (OR 1.37, 95% CI 0.89-2.12), moderately severe depressive symptoms (OR 1.93, 95% CI 0.93-3.99), and severe depressive symptoms (OR 0.67, 95% CI 0.16-2.88).As a continuous variable, life-ever gallstones was associated with the PHQ-9 score (OR 0.42, 95% CI 0.02-0.83). The results remained stable after multiple imputation for all missing data. This cross-sectional study demonstrates no significant association between life-ever gallstones and depressive symptoms in US adults.

Computational underwater ghost imaging based on scattering-and-absorption degradation.

In underwater computational ghost imaging, the presence of scattering and absorption introduces significant degradation, leading to blurring and distortion of illuminating patterns. This work proposes an anti-degradation underwater computational ghost imaging (AUGI) method based on the physical degradation model of underwater forward degradation caused by scattering and absorption. Through AUGI, we can enhance the quality of a reconstructed image by about 10% compared to differential ghost imaging (DGI) as measured by peak signal-to-noise ratio (PSNR) and structural similarity (SSIM), as a result of simulations. We experimentally demonstrate the superior effectiveness of this method in the artificial submarine environment. Additionally, benefitting from its simplicity, this method is expected to be applied across a wide range of underwater ghost imaging applications.

Corrigendum: Trends in acupuncture for infertility: a scoping review with bibliometric and visual analysis.

[This corrects the article DOI: 10.3389/fendo.2024.1351281.].

Prevalence, cessation, and geographical variation of smoking among middle-aged and elderly adults in China: A population-based study.

INTRODUCTION: Smoking significantly burdens human health, contributing to an increasing incidence of mortality and morbidity. This study aims to explore the prevalence of smoking, cessation, and the association between various risk factors and smoking intensity measured in pack-years among Chinese adults. METHODS: During 2020-2021, the China Stroke High-risk Population Screening and Intervention Program (CSHPSIP) invited participants aged ≥40 years from 31 provinces in mainland China. This cross-sectional study presents the standardized prevalence of smoking and cessation across various demographics, including age, sex, residence, income, education level, BMI, and geographical region of residence. Multivariable logistic regression was used to examine the associations between smoking pack-years and related factors. RESULTS: Among 524741 participants (mean age: 61.9 ± 10.9 years; 41.1% male; 58.9% female), standardized smoking prevalence was 19.3% (95% CI: 19.2-19.4), with men (37.2%; 95% CI: 37.0-37.4) displaying significantly higher rates than women (1.3%; 95% CI: 1.2-1.3). Smoking cessation rate stood at 11.2% (95% CI: 11.0-11.4), with 11.3% (95% CI: 11.1-11.5) for men and 8.4% (95% CI: 7.5-9.2) for women. Urban residents and those with advanced education had lower smoking rates and higher cessation rates. Additionally, the dose-response relationship indicated a more pronounced association between higher smoking pack-years and elevated health risks, including hypertension (AOR=1.30; 95% CI: 1.24-1.36), diabetes (AOR=1.26; 95% CI: 1.20-1.33), hyperlipidemia (AOR=1.22; 95% CI: 1.16-1.28), heart disease (AOR=1.40; 95% CI: 1.26-1.54), and stroke (AOR=1.23; 95% CI: 1.10-1.36). CONCLUSIONS: This comprehensive study emphasizes the profound impact of smoking on health in Chinese adults, indicating the critical need for tailored cessation programs, particularly for middle-aged individuals, men, rural residents, and those with lower level of education.

A pair of E3 ubiquitin ligases control immunity and flowering by targeting different ELF3 proteins in rice.

The ubiquitin-proteasome system (UPS) plays crucial roles in cellular processes including plant growth, development, and stress responses. In this study, we report that a pair of E3 ubiquitin ligases, AvrPiz-t-interaction protein 6 (APIP6) and IPA1-interaction protein 1 (IPI1), intricately target early flowering3 (ELF3) paralogous proteins to control rice immunity and flowering. APIP6 forms homo-oligomers or hetero-oligomers with IPI1. Both proteins interact with OsELF3-2, promoting its degradation to positively control resistance against the rice blast fungus (Magnaporthe oryzae). Intriguingly, overexpression of IPI1 in Nipponbare caused significantly late-flowering phenotypes similar to the oself3-1 mutant. Except for late flowering, oself3-1 enhances resistance against M. oryzae. IPI1 also interacts with and promotes the degradation of OsELF3-1, a paralog of OsELF3-2. Notably, IPI1 and APIP6 synergistically modulate OsELF3s degradation, finely tuning blast disease resistance by targeting OsELF3-2, while IPI1 controls both disease resistance and flowering by targeting OsELF3-1. This study unravels multiple functions for a pair of E3 ligases in rice.

Forsythoside B alleviates cerebral ischemia-reperfusion injury via inhibiting NLRP3 inflammasome mediated by SIRT1 activation.

BACKGROUND: The inflammatory response is a key factor in the pathogenesis of cerebral ischemia/reperfusion injury (CIRI), and anti-inflammatory interventions may offer a promising therapeutic strategy. Forsythoside B (FB) is a phenylethanoid glycoside isolated from Forsythiae fructus, which has been reported to have anti-inflammatory effects. However, the mechanism of the neuroprotective effect of FB on CIRI remains unclear. METHODS: Adult male Sprague-Dawley rats were subjected to transient middle cerebral artery occlusion/reperfusion (MCAO/R). FB was administered intraperitoneally for 3 days prior to MCAO/R. Cerebral infarct volume and neurological deficit score were used as indices to evaluate MCAO/R injury. The serum levels of inflammatory factors and antioxidant enzymes were measured. The activation of silent information regulator 2 homolog 1 (Sirt1) and the inhibition of the nucleotide-binding oligomerization domain-like receptor with a pyrin domain 3 (NLRP3) pathway were assessed through western blot and immunohistochemistry analysis. Furthermore, the rats were treated with Sirt1 shRNA 3 days before MCAO/R by stereotactical injection into the ipsilateral hemispheric region to assess the impact of Sirt1 knockdown on the protection of FB during MCAO/R. RESULTS: FB reduced cerebral infarct volume and neurological deficit score in MCAO/R rats. FB reduced pathological changes and cell apoptosis in the hippocampal CA1 region and cortex on the ischemic side of rats. FB inhibited the serum levels of inflammatory factors and increased the activities of antioxidant enzymes. Further study showed that FB inhibited the activation of the NLRP3 pathway and induced Sirt1 activation. CONCLUSION: FB demonstrated neuroprotective and anti-inflammatory effects by inhibiting the NLRP3 pathway through Sirt1 activation in CIRI.

[Prophylaxis of migraine without aura of liver yang hyperactivity treated with acupuncture at the points selected using the "seven lines of the neck" method: a randomized controlled trial].

OBJECTIVE: To observe the clinical effect of prophylaxis on migraine without aura differentiated as liver yang hyperactivity undergoing acupuncture at the points selected using the "seven lines of the neck" method. METHODS: Fifty-eight patients with migraine without aura of liver yang hyperactivity at remission stage were randomly divided into an observation group (29 cases, 3 cases dropped out) and a control group (29 cases, 4 cases dropped out). In the observation group, acupuncture was delivered at Dazhui (GV 14), Fengfu (GV 16), bilateral Fengchi (GB 20), Gongxue (Extra), etc., selected using the "seven lines of the neck" method. In the control group, conventional acupuncture was applied to ashi point, Sizhukong (TE 23), Shuaigu (GB 8), Taiyang (EX-HN 5) and others on the affected side. The treatment was given once every other day or every two days, 3 interventions weekly, for consecutive 8 weeks. Before treatment, after 4 and 8 weeks of treatment, and after 4 weeks of treatment completion (follow-up visit), the days of migraine episodes, the frequency of migraine episodes, the score of visual analogue scale (VAS) for pain intensity, and the score of migraine specific quality of life questionnaire (MSQ) were observed in the patients of the two groups. Before treatment and after 8 weeks of treatment, the score of TCM syndrome was observed. After 4 and 8 weeks of treatment and after 4 weeks of treatment completion (follow-up visit), the response rates of 50% reduction in the days and the frequency of migraine episodes were calculated in the two groups. RESULTS: After 4 and 8 weeks of treatment and during follow-up visit, the days and the frequency of migraine episodes were decreased (P<0.01) and VAS scores were declined (P<0.01) when compared with those before treatment in the two groups. The days and the frequency of migraine episodes in the observation group were lower during the follow-up visit (P<0.05) and VAS scores were lower after 8 weeks of treatment and during the follow-up visit (P<0.05) when compared with those in the control group. After 4 and 8 weeks of treatment, and during follow-up visit, the scores of "role function-preventive" and "emotional function" of MSQ were increased in comparison with those before treatment in the observation group (P<0.05). After 8 weeks of treatment and during the follow-up visit, the scores of "role function-restrictive" of MSQ were increased in comparison with those before treatment in the observation group (P<0.05), and the scores of "role function-restrictive" "role function-preventive" and "emotional function" were higher when compared with those before treatment in the control group (P<0.05). After 8 weeks of treatment, the scores of TCM syndrome were decreased in comparison with those before treatment in the two groups (P<0.01). In the observation group, the response rate of 50% reduction in the days of migraine episodes after 8 weeks of treatment and that of the frequency of migraine episodes during the follow-up visit were higher than those of the control group (P<0.05). CONCLUSION: Acupuncture at the points selected using the "seven lines of the neck" method can reduce the days and frequency of migraine episodes and pain intensity, ameliorate the syndrome of TCM and improve the quality of life of the patients with migraine without aura of liver yang hyperactivity.

Trends in acupuncture for infertility: a scoping review with bibliometric and visual analysis.

Background: Unexplained recurrent implantation failure and the high cost of assisted reproductive techniques for those experiencing infertility have increasingly resulted in the use of acupuncture. However, the trends and research status of acupuncture on infertility resulting in natural conception have not been systematically summarized. This scoping review and knowledge graph analysis aimed to summarize existing clinical studies on acupuncture for infertility that resulted in natural conception. Methods: Seven databases, namely, PubMed, Embase, the Cochrane Library, CNKI, VIP, Wanfang Data, and SinoMed, were searched up to August 2023 (updated on 1 April). Two authors independently identified related clinical studies and systematic reviews, and extracted data from included studies on acupuncture for infertility; any discrepancies were resolved by discussion or judged by a third author. A meta-analysis was conducted based on randomized controlled trials (RCTs), and data were synthesized using risk ratios with 95% confidence intervals. Results: Of the 310 articles meeting the inclusion criteria, 274 were primary studies, 7 were systematic reviews, and 29 were case reports. Reported adverse events included mild ovarian irritation and early signs of miscarriage. Out of the 274 primary studies, there were 40 (14.60%) cases of male infertility and 234 (85.40%) cases of female infertility. Current research highlights on acupuncture for infertility focused on female infertility caused by polycystic ovary syndrome, ovulation disorder, and luteinized unruptured follicle syndrome (LUFS), while acupuncture for male infertility was a hotspot in the early research stage. The meta-analysis also suggested that acupuncture was more effective than human chorionic gonadotropin (HCG) [RR = 1.89, 95% CI (1.47, 2.42), 11 RCTs, 662 participants]. Acupuncture combined with HCG was comparable to HCG [RR = 2.33, 95% CI (1.53, 3.55), four RCTs, 259 participants]. Compared with no treatment, acupuncture resulted in a higher pregnancy rate [RR = 22.12, 95% CI (1.39, 353.09), one RCT, 47 participants]. There was no statistical difference between acupuncture combined with HCG plus letrozole and HCG plus letrozole [RR = 1.56, 95% CI (0.84, 2.89), one RCT, 84 participants]. Conclusion: Current research highlights on acupuncture for infertility resulting in natural conception focused on female infertility caused by polycystic ovary syndrome, ovulation disorder, and LUFS, while studies on male infertility and female infertility caused by blockage in the fallopian tube, thin endometrium, and other factors were insufficient. Well-designed confirmatory clinical studies are still needed as the research hypotheses of most studies were unclear.

Progress of researches on mechanisms of acupuncture therapy in the treatment of lumbar disc herniation. / é’ˆç¸æ²»ç–—è °æ¤Žé—´ç›˜çªå‡ºç—‡ä½œç”¨æœºåˆ¶ç ”ç©¶è¿›å±•.

Lumbar intervertebral disc herniation (LDH) is a common and frequently-occurring disease, which usually causes lumbar and leg pain. Studies have shown that acupuncture can improve the symptoms of LDH patients. In the present paper, we summarize the progress of researches on the mechanisms of acupuncture underlying improvement of symptoms of LDH in recent 10 years from 1) delaying the intervertibral disc degeneration (by down-regulating the expressions of matrix metalloproteinase ï¼»MMPï¼½-3 and MMP-4, up-regulating the expressions of diosaccharides and polyglycoprotein, inhibiting apoptosis and promoting mitochondrial autophagy of nucleus pulposus cells, etc.), 2) maintaining spinal column stability (by relieving rachiasmus and improving lumbar flexor and extensor muscle strength, lowering the degree of polyfidus edema and fat infiltration, and restoring the biomechanics of the spine), 3) regulating inflammation (by inhibiting the production of proinflammatory factors and increasing the production of anti-inflammatory factors, etc.), 4) regulating immune response (by promoting the activity of T cells and other immune cells, lowering serum levels of MMP-3, transforming growth factor-ß1 and prostaglandin E2, raising serum levels of IgA, IgG and IgM to improve immune function ), 5) modulating neural structure and function (by promoting myelin regeneration of sciatic nerve fibers, and reducing the edema of Schwann cells' cytoplasm and mitochondria, and improving neural ultrastructure, and sensory and motor functions of peripheral nerves, etc.), 6) relieving lumbar pain (by down-regulating expression of Ca2+/calmodulin-dependent protein kinase and activation of lumbar spinal cord glial cells, blocking nociceptive signal conduction, regulating the levels of pain-related factors, etc.), and 7) improving local microcirculation. These results may provide scientific evidence for acupuncture treatment of LDH.

Research on traditional Chinese medicine as an effective drug for promoting wound healing.

ETHNOPHARMACOLOGICAL RELEVANCE: The incidence of skin trauma is high and the repair process is complex, often leading to poor healing and other issues, which can result in significant economic and social burdens. Traditional Chinese medicine (TCM) is a valuable resource with proven effectiveness and safety in wound repair, widely utilized in clinical practice. A systematic analysis of wound healing with a focus on TCM research progress holds both academic and clinical importance. AIM OF THE REVIEW: This article reviews the research progress of TCM in promoting wound healing, and provides basic data for the development of innovative drugs that promote wound healing. MATERIALS AND METHODS: This article provides a review of the literature from the past decade and conducts a thorough analysis of various databases that contain reports on the use of TCM for wound repair. The data for this systematic research was gathered from electronic databases including CNKI, SciFinder, and PubMed. The study explores and summarizes the research findings and patterns by creating relevant charts. RESULTS: This study reviewed the mechanism of wound healing, experimental TCM methods to promote wound healing, the theory and mode of action of TCM to promote wound healing, the active ingredients of TCM that promote wound healing, the efficacy of TCM formulae to promote wound healing, and the potential toxicity of TCM and its antidotes. This study enriched the theory of TCM in promoting wound healing. CONCLUSION: Skin wound healing is a complex process that can be influenced by various internal and external factors. This article offers a theoretical foundation for exploring and utilizing TCM resources that enhance wound repair. By analyzing a range of TCM that promote wound healing, the article highlights the clinical importance and future potential of these medicines in promoting wound healing.

The association between urinary incontinence and suicidal ideation: Findings from the National Health and Nutrition Examination Survey.

BACKGROUND: Urinary incontinence (UI) might be linked to suicidal ideation, but we do not yet have all the relevant details. This study aimed to dig deeper into the connection between UI and suicidal ideation using data from the National Health and Nutrition Examination Survey (NHANES). METHODS: We examined 31,891 participants aged ≥ 20 years from NHANES 2005-2018 who provided complete information. We used standardized surveys to check for UI and signs of suicidal ideation. To better understand this relationship, we used statistical tools such as multivariable logistic regression, subgroup analysis, and sensitivity analyses. RESULTS: Among the 31,891 participants, 28.9% reported UI and 10.7% reported suicidal ideation. Those with UI exhibited a significantly greater incidence of suicidal ideation (15.5%) than did those without UI (8.8%, P < 0.001). After adjusting for various factors, including age, sex, marital status, socioeconomic status, educational level, lifestyle factors, and chronic comorbidities, UI remained significantly associated with suicidal ideation (OR:1.54, 95% CI = 1.39-1.7, P < 0.001). Among all types of UI, MUI participants were more likely to experience suicidal ideation. Compared with no UI, higher odds of suicidal ideation suffered from MUI (OR:2.11, 95%CI:1.83-2.44, P < 0.001), SUI (OR:1.4, 95%CI:1.19-1.65, P < 0.001), UUI(OR:1.37,95%CI:1.16-1.62, P < 0.001) after full adjustment. With the exception of individuals living with a partner, the remaining subgroups exhibited a positive correlation between urinary incontinence and suicidal ideation, considering that factors such as age, sex, and prevalent comorbidities such as hypertension, depression, and diabetes did not reveal any statistically significant interactions (all P > 0.05). Sensitivity analyses, incorporating imputed missing covariates, did not substantially alter the results (OR: 1.53, 95% CI: 1.4-1.68, P < 0.001). CONCLUSION: Urinary incontinence may correlate with increased suicidal ideation risk, priority screening for suicidal ideation and timely intervention are essential for individuals with urinary incontinence, but prospective studies are needed to verify the results.

The rice E3 ubiquitin ligase-transcription factor module targets two trypsin inhibitors to enhance broad-spectrum disease resistance.

Broad-spectrum disease resistance (BSR) is crucial for controlling plant diseases and relies on immune signals that are subject to transcriptional and post-translational regulation. How plants integrate and coordinate these signals remains unclear. We show here that the rice really interesting new gene (RING)-type E3 ubiquitin ligase OsRING113 targets APIP5, a negative regulator of plant immunity and programmed cell death (PCD), for 26S proteasomal degradation. The osring113 mutants in Nipponbare exhibited decreased BSR, while the overexpressing OsRING113 plants showed enhanced BSR against Magnaporthe oryzae (M. oryzae) and Xanthomonas oryzae pv. oryzae (Xoo). Furthermore, APIP5 directly suppressed the transcription of the Bowman-Birk trypsin inhibitor genes OsBBTI5 and AvrPiz-t-interacting protein 4 (APIP4). Overexpression of these two genes, which are partially required for APIP5-mediated PCD and disease resistance, conferred BSR. OsBBTI5 and APIP4 associated with and stabilized the pathogenesis-related protein OsPR1aL, which promotes M. oryzae resistance. Our results identify an immune module with integrated and coordinated hierarchical regulations that confer BSR in plants.

[Application of high flow nasal canula in patients with pulmonary edema caused by seawater drowning].

OBJECTIVE: To investigate the therapeutic effect of high-flow nasal cannula oxygen therapy (HFNC) and non-invasive positive pressure ventilation (NPPV) on patients with pulmonary edema caused by seawater drowning. METHODS: A retrospective analysis method was used. Based on the Utstein database of emergency drowning in the First Hospital of Qinhuangdao, the clinical data of patients with seawater drowning pulmonary edema admitted to the emergency medicine department of the First Hospital of Qinhuangdao from January 1, 2019 to December 31, 2022 were collected. The patients were divided into NPPV group and HFNC group according to different ventilation methods. The general data, endotracheal intubation rate in 7 days, arterial blood gas analysis indexes [arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), arterial oxygen saturation (SaO2)] and hemodynamic indexes (systolic blood pressure, diastolic blood pressure, mean arterial pressure, heart rate, blood lactic acid) before and after treatment, length of stay in intensive care unit (ICU), oxygen therapy comfort of the two groups were compared. RESULTS: A total of 54 patients were enrolled, including 21 patients in the NPPV group and 33 patients in the HFNC group. There were no significant differences in gender, age, state of consciousness and other general information between the two groups. Compared with NPPV group, the rate of endotracheal intubation in HFNC group within 7 days was significantly lower [24.2% (8/33) vs. 33.3% (7/21), P < 0.05]. Before treatment, there were no significant differences in arterial blood gas analysis and hemodynamics between the two groups. After treatment, the above indexes in both groups were significantly improved compared with those before treatment, and PaO2, SaO2, systolic blood pressure, diastolic blood pressure and mean arterial pressure in HFNC group were significantly higher than those in NPPV group [PaO2 (mmHg, 1 mmHg≈0.133kPa): 93.56±6.37 vs. 82.14±6.25, SaO2: 1.02±0.09 vs. 0.95±0.11, systolic blood pressure (mmHg): 117.37±8.43 vs. 110.42±8.38, diastolic blood pressure (mmHg): 79.43±7.61 vs. 72.21±4.32, mean arterial pressure (mmHg): 92.34±6.32 vs. 85.12±5.38], PaCO2, heart rate and blood lactic acid were significantly lower than those in NPPV group [PaCO2 (mmHg) : 34.26±5.63 vs. 37.24±6.22, heart rate (times/min): 73.38±7.56 vs. 86.25±5.41, blood lactic acid (mmol/L): 1.38±0.36 vs. 2.25±1.14], and the differences were statistically significant (all P < 0.05). In addition, the length of ICU stay in HFNC group was significantly shorter than that in NPPV group (days: 13.30±2.38 vs. 16.27±4.26), and the comfort rate of oxygen therapy was significantly higher than that in NPPV group [66.7% (22/33) vs. 42.8% (9/21)], with statistical significance (all P < 0.05). CONCLUSIONS: HFNC can improve the oxygenation of patients with pulmonary edema caused by seawater drowning, improve hemodynamics, reduce the rate of tracheal intubation, shorten the length of ICU stay, and improve the comfort of oxygen therapy, which has certain clinical application value.

Rapid and ultra-sensitive trace metals detection of water by partial Leidenfrost superhydrophobic array surface enhanced laser-induced breakdown spectroscopy.

The rapid and ultra-sensitive detection of trace elements in liquid is a primary concern for researchers. In this study, a partial Leidenfrost effect superhydrophobic (PLSHB) array surface was used for rapid in situ evaporation enrichment of sample droplets. Within 4 min, a 50 µL droplet sample was completely evaporated, resulting in all solutes in it being concentrated within a circular range measuring approximately 350 µm in diameter, without the formation of a coffee ring structure. The limits of detection for six metals (Pb, Ba, Be, Mn, Cr, Cu) in water were determined to be as follows: 0.82 µgL-1, 0.27 µgL-1, 0.033 µgL-1, 0.136 µgL-1, 0.241 µgL-1, and 0.083 µgL-1. Furthermore, laser-induced breakdown spectroscopy (LIBS) was employed to detect the enriched solutes from ten liquid samples with identical concentrations on the PLSHB array surface; these measurements exhibited a relative standard deviation (RSD) of only 3.7%. Spike experiments involving the addition of the aforementioned six metals into drinking water demonstrated recovery rates ranging from 85.7% to 117.7%. Therefore, the application potential of PLSHB array surface enhanced LIBS for rapid, stable, and ultra-sensitive detection and analysis of trace metal elements across various fields such as industry, environmental science, and biomedicine might be highly promising.

MecVax supplemented with CFA MEFA-II induces functional antibodies against 12 adhesins (CFA/I, CS1-CS7, CS12, CS14, CS17, and CS21) and 2 toxins (STa, LT) of enterotoxigenic Escherichia coli (ETEC).

Enterotoxigenic Escherichia coli (ETEC) strains that produce various adhesins and one or two enterotoxins are the leading causes of children's diarrhea and travelers' diarrhea. MecVax, a multivalent ETEC vaccine candidate, consists of two proteins, an adhesin multiepitope fusion antigen (MEFA) that stimulates antibodies to the seven most important ETEC adhesins (CFA/I and CS1-CS6) and a toxoid fusion antigen which stimulates antibodies against ETEC enterotoxins (heat-labile toxin and heat-stable toxin). CFA MEFA-II, another polyvalent MEFA protein, has been demonstrated to stimulate antibodies to another five important ETEC adhesins (CS7, CS12, CS14, CS17, and CS21). We hypothesize that MecVax coverage and efficacy can be expanded if MecVax could stimulate antibodies to all 12 adhesins. In this study, we supplemented MecVax with CFA MEFA-II, examined broad immunity to the 12 targeted ETEC adhesins and 2 ETEC toxins (STa, LT) in mice, and assessed mouse antibody functions for inhibiting the adherence of the 12 adhesins and neutralizing the enterotoxicity of 2 toxins, thus assessing the potential application of a broadly protective pan-ETEC vaccine. Mice intramuscularly immunized with MecVax and CFA MEFA-II developed robust antibody responses to the 12 ETEC adhesins and 2 toxins; furthermore, mouse serum antibodies showed functional activities against the adherence from each of the targeted adhesins and the enterotoxicity of either toxin. Data also indicated that CFA MEFA-II was antigenically compatible with MecVax. These results demonstrated that the inclusion of CFA MEFA-II further expands MecVax broad immunogenicity and protection coverage, suggesting the feasibility of developing a vaccine against all important diarrheal ETEC strains.IMPORTANCEThere are no vaccines licensed for Enterotoxigenic Escherichia coli (ETEC), a leading cause of children's diarrhea and the most common cause of travelers' diarrhea. Since ETEC strains produce over 25 adhesins and 2 distinctive enterotoxins, heterogeneity is a key obstacle to vaccine development. MecVax, a multivalent ETEC vaccine candidate, induces protective antibodies against the seven most important adhesins (CFA/I and CS1-CS6) associated with two-thirds of ETEC clinical cases. However, ETEC prevalence shifts chronically and geographically, and other adhesins are also associated with clinical cases. MecVax would become a pan-ETEC vaccine if it also protects against the remaining important adhesins. This study demonstrated that MecVax supplemented with adhesin protein CFA MEFA-II induces functional antibodies against 12 important ETEC adhesins (CFA/I, CS1-CS7, CS12, CS14, CS17, and CS21), enabling the development of a more broadly protective ETEC vaccine and further validating the application of the MEFA vaccinology platform for multivalent vaccine development.