Little is known about the association between the COVID‐19 pandemic and early survival among newly diagnosed cancer patients.This retrospective population‐based cohort study used linked administrative datasets from Ontario, Canada. Adults (≥18 years) who received a cancer diagnosis between March 15 and December 31, 2020, were included in a pandemic cohort, while those diagnosed during the same dates in 2018/2019 were included in a pre‐pandemic cohort. All patients were followed for one full year after the date of diagnosis. Cox proportional hazards regression models were used to assess survival in relation to the pandemic, patient characteristics at diagnosis, and the modality of first cancer treatment as a time‐varying covariate. Interaction terms were explored to measure the pandemic association with survival for each cancer type.Among 179,746 patients, 53,387 (29.7%) were in the pandemic cohort and 37,741 (21.0%) died over the first post‐diagnosis year. No association between the pandemic and survival was found when adjusting for patient characteristics at diagnosis (HR 0.99 [95% CI 0.96–1.01]), while marginally better survival was found for the pandemic cohort when the modality of treatment was additionally considered (HR 0.97 [95% CI 0.95–0.99]). When examining each cancer type, only a new melanoma diagnosis was associated with a worse survival in the pandemic cohort (HR 1.25 [95% CI 1.05–1.49]).Among patients able to receive a cancer diagnosis during the pandemic, one‐year overall survival was not different than those diagnosed in the previous 2 years. This study highlights the complex nature of the COVID‐19 pandemic impact on cancer care. This population‐based cohort study in Ontario, Canada found no association between the pandemic and the 1‐year overall survival of adults newly diagnosed with cancer. These results highlight the complex nature of the pandemic impact on cancer care. Administrative datasets were linked using unique encoded identifiers and analyzed at ICES. Ethical approval was not sought, as the use of data was approved by ICES' Privacy and Legal Office. This study followed the Reporting of studies Conducted using Observational Routinely collected health data (RECORD) and the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) guidelines. The analysis was conducted between June–July 2022.2.2The Ontario Cancer Registry (OCR) is a population‐based registry capturing 96% of cancer diagnoses in the province.2.3n = 1624, 0.9%) to mitigate the impact of an expected drop in the number of patients whose cancer diagnosis was documented by death certificate during the pandemic; a sensitivity analysis was later performed to further examine this issue. The date of death was obtained from the RPDB, a validated source of vital statistics that was updated until April 2022 at the time of this analysis. For patients in the final cohort, they were followed from the date of cancer diagnosis for 1 year (365 days) or until the date of death from any cause, whichever happened first. Hence, a complete 1‐year follow‐up window was available for each patient.We created a pandemic cohort, consisting of adults (≥18 years) with an invasive cancer diagnosis from March 15 to December 31, 2020, and a pre‐pandemic cohort, consisting of those diagnosed during the corresponding periods in 2019 and 2018. March 15, 2020, was when hospitals in Ontario were advised by the Chief Medical Officer of Health to discontinue nonemergent and elective procedures.2.4The outcome of this study was overall survival within 1 year of cancer diagnosis, a time‐to‐event variable capturing the number of days from cancer diagnosis (index date) to death. Patients who were alive at the end of the first year after diagnosis were censored at that point.2.5The following patient characteristics were measured at the time of cancer diagnosis. Age and sex were obtained from the RPDB. Rural residence was defined as living in a small town or rural area with a population of less than 10,000 in the PCCF.For each patient, we followed an established algorithm to determine the modality of first cancer treatment: surgery, systemic therapy, radiotherapy, or being untreated over the first post‐diagnosis year.2.6p‐value<0.05. Analyses were performed on SAS Enterprise Guide 7.15 (SAS Institute).We compared the characteristics of the pre‐pandemic and the pandemic cohort using 0.10 as a standardized difference threshold.3n = 135,828, 75.6%), the mean time‐to‐treatment was 6 days shorter in the pandemic cohort, dropping from 51.5 days (SD 57.5) to 45.6 days .Of the 179,746 patients included, 126,359 (70.3%) patients were in the pre‐pandemic cohort and 53,387 (29.7%) were in the pandemic cohort. At the time of cancer diagnosis, there were no significant differences in sociodemographic and clinical characteristics between the two cohorts . Specifically, the estimated 1‐year survival probability was 78.7% (95% CI: 78.4%–79.1%) for the pandemic cohort, compared to 78.1% (95% CI: 77.9%–78.3%) for the pre‐pandemic cohort.A full 1‐year of data was available on all patients for capturing mortality. At the end of the first year after diagnosis, 37,741 (21.0%) patients in the entire cohort died. The Kaplan–Meier plot . Specifically, hepatobiliary  and lung  cancers were associated with a higher survival for the pandemic cohort, while melanoma was associated with a worse survival for the pandemic cohort .Interaction between the pandemic and each cancer type was added to Model 2 (Model 3). In Appendix n = 181,370) now included those who died on the date of cancer diagnosis (n = 1624). Again, we observed no difference in patient characteristics at cancer diagnosis between the two cohorts . The higher survival persisted in all three Cox models  those diagnosed later in the pandemic continued to demonstrate stable early survival.Our analysis was subject to the usual limitations of administrative data in being able to control for confounders, particularly the lack of cancer staging data, which typically require 2–3 years after diagnosis to mature within our cancer registry. Existing literature from the Unites States, Australia, and Europe suggested the pandemic‐associated upshift in cancer stage may be smaller than expected5We used linked administrative data from a large province wide population‐based cohort to assess overall survival over the first year among adults diagnosed with cancer during the pandemic  and in pre‐pandemic . Overall, no difference in 1‐year survival was found for patients diagnosed during the pandemic. Individually, a small but significantly higher survival was observed for hepatobiliary and lung cancers. Melanoma was the only cancer type associated with worse survival during the pandemic. These findings demonstrate the complex effect of the pandemic on the trajectory of cancer care that may not have necessarily translated into a decrease in early survival for those diagnosed early with COVID‐19. Efforts should be made to find the “missing” cancer patients and closely monitor the status of existing patients.Rui Fu: Conceptualization ; investigation ; methodology ; writing – original draft (lead); writing – review and editing . Rinku Sutradhar: Conceptualization ; data curation ; investigation ; methodology ; supervision ; writing – review and editing . Qing Li: Conceptualization ; data curation ; formal analysis ; methodology ; writing – review and editing . Pabiththa Kamalraj: Conceptualization ; investigation ; project administration ; writing – review and editing . Anna Dare: Conceptualization ; investigation ; writing – review and editing . Timothy Hanna: Conceptualization ; investigation ; writing – review and editing . Kelvin Chan: Conceptualization ; investigation ; writing – review and editing . Jonathan C Irish: Conceptualization ; investigation ; writing – review and editing . Natalie Coburn: Conceptualization ; investigation ; writing – review and editing . Juile Hallet: Conceptualization ; investigation ; writing – review and editing . Simron Singh: Conceptualization ; investigation ; writing – review and editing . Ambica Parmar: Conceptualization ; investigation ; writing – review and editing . Craig C Earle: Conceptualization ; investigation ; writing – review and editing . Lauren Lapointe‐Shaw: Conceptualization ; investigation ; writing – review and editing . Monika Krzyzanowska: Conceptualization ; investigation ; writing – review and editing . Alexander V. Louie: Conceptualization ; investigation ; writing – review and editing . Alyson Mahar: Conceptualization ; investigation ; methodology ; writing – review and editing . David Urbach: Conceptualization ; investigation ; writing – review and editing . Daniel McIsaac: Conceptualization ; investigation ; writing – review and editing . Danny J. Enepekides: Conceptualization ; investigation ; writing – review and editing . David Gomez: Conceptualization ; investigation ; writing – review and editing . Nicole J Look Hong: Conceptualization ; investigation ; writing – review and editing . Jill Tinmouth: Conceptualization ; investigation ; methodology ; writing – review and editing . Antoine Eskander: Conceptualization ; funding acquisition ; investigation ; methodology ; supervision ; writing – review and editing .This study was supported by ICES, which is funded by an annual grant from the Ontario Ministry of Health and the Ministry of Long‐Term Care. This study also received funding from a Sunnybrook Research Institute and Sunnybrook Foundation COVID‐19 Response Grant and a Canadian Institutes of Health Research Operating Grant #179892. The funders had no role in the design and conduct of the study, nor the decision to prepare and submit the manuscript for publication.Dr. Antoine Eskander reports research funding from Merck (2019) and consultant fee from Bristol‐Myers Squibb (2019). All other listed authors have disclosed that they have not received any financial consideration from any person or organization to support the preparation, analysis, results, or discussion of this article.Data S1:Click here for additional data file.


Cucumeropsis mannii seed oil (CMSO) capable of alleviating BPA‐induced dyslipidemia and adipokine dysfunction. In this study, we evaluated the effects of CMSO on adipokine dysfunctions and dyslipidemia in bisphenol‐A (BPA)‐induced male Wistar rats. Six‐week‐old 36 albino rats of 100–200 g weight were assigned randomly to six groups, which received varied doses of BPA and/or CMSO. The administration of BPA and CMSO was done at the same time for 42 days by oral intubation. The adipokine levels and lipid profile were measured in adipose tissue and plasma using standard methods. BPA induced significant (p < .05) increases in triglycerides, cholesterol, leptin, LDL‐C, and atherogenic and coronary risk indices in adipose tissue and plasma, as well as a decrease in adiponectin and HDL‐C levels in Group II animals. BPA administration significantly (p < .05) elevated Leptin levels and reduced adiponectin levels. BPA plus CMSO reduced triglycerides, cholesterol, leptin, LDL‐C, and atherogenic and coronary risk indices while increasing adiponectin levels and HDL‐C in adipose tissue and plasma (p < .05). The results showed that BPA exposure increased adipose tissue as well as serum levels of the atherogenic index, triglycerides, cholesterol, coronary risk index, LDL‐C, leptin, and body weight with decreased adiponectin levels and HDL‐C. Treatment with CMSO reduced the toxicities caused by BPA in rats by modulating the body weight, adiponectin/leptin levels, and lipid profiles in serum and adipose tissue. This study has shown that CMSO ameliorates BPA‐induced dyslipidemia and adipokine dysfunctions. We suggest for further clinical trial to establish the clinical applications.This study demonstrated the therapeutic potentials of  p < .05). Treatment with CMSO reduced the toxicities caused by BPA in rats by modulating the body weight, adiponectin and leptin levels, and lipid profiles in adipose tissue and serum.BPA plus CMSO reduced triglycerides, cholesterol, leptin, LDL‐C, and atherogenic and coronary risk indices while increasing adiponectin levels and HDL‐C in adipose tissue and plasma ( The obtained plant was identified by Mr. Nwankwo, E.O., a taxonomist at Ebonyi State University, Nigeria (EBSU). For the purpose of reference, a portion of the C. mannii seed was maintained at the herbarium of EBSU with voucher Reference number: EBSU‐H‐396.2.3CMSO was extracted by applying the traditional and mechanical methods of Kate et al.  and modi2.4CMSO acute toxicity in Wistar Albino rats  was evaluated according to OECD  of 100–200 g weight, which were randomly grouped into six groups labeled groups I to VI, with each group having six rats. Groups I to III were control, whereas groups IV to VI were treated groups. Groups I, II, and III were given 1 ml olive oil, 100 mg/kg bw BPA, and 7.5 ml/kg bw CMSO, respectively. Groups IV, V, and VI received 100 mg/kg bw BPA + 7.5 mg/kg bw CMSO, 5 mg/kg bw CMSO, and 2.5 mg/kg bw CMSO, respectively. Body weight was measured every 7 days during the trials. The CMSO and the BPA were simultaneously administered using oral intubation for 6 weeks, and done once per day  with Brown–Forsythe test was done. Generally, p < .05 (significance level).The statistical analyses were done with GraphPad Prism 5.04. The data/results were presented as mean standard deviation. 33.1p < .05) elevated the rat weights  decrease in the body weight of rats after coadministration of BPA + CSMO  elevated the serum levels of cholesterol and LDL‐C with a reduction in HDL‐C levels. However, BPA and CMSO coadministration significantly (p < .05) reduced cholesterol, triglycerides, and LDL‐C with an elevation of HDL‐C. A nonsignificant(p > .05) increase was observed in the level of triglycerides in normal control when compared with the BPA control. But coadministration of BPA and CMSO significantly(p < .05) elevated the triglycerides level in serum  elevated the levels of cholesterol, triglycerides, and LDL‐C with a reduction in HDL‐C level in rat adipose tissue. However, BPA and CMSO coadministration significantly (p < .05) reduced cholesterol, triglycerides, and LDL‐C with the elevation of HDL‐C  elevated the CRI in serum and adipose tissue of male rats  reduced the CRI in both adipose tissue and serum of rats  elevated the ARI in adipose tissue and serum of male rats  reduced the ARI in both adipose tissue and serum of rats  elevated leptin level and reduced adiponectin level in adipose tissues of male rats  reduced the level of leptin but elevated the adiponectin level in male rats  elevated leptin levels and reduced adiponectin levels in male rats' serum  reduced the level of leptin though not in a dose‐dependent manner and elevated the adiponectin level in male rats  and prevents it from catalyzing the conversion of 3‐hydroxy‐3‐methylglutaryl‐coenzyme‐A to mevalonic acid, a critical step in biosynthesis  increased serum and adipose tissue levels of leptin while concurrently reducing levels of adiponectin. The findings of this study matched those of Angle et al.  and delivered to organs and tissues throughout the body, or fatty acids are released from adipocytes and mobilized for use during times of stress when the body requires energy . In an earlier study  injection increased the coronary risk index and atherogenic index (AI) in the serum and adipose tissue of male rats considerably (This finding coincided with an animal study conducted in the United State of America, which found that BPA injection in rats increased AI and CRI in serum and adipose tissues above normal ranges and was inversely associated with BMI (Iwaniec et al., The findings of this work are comparable to the study of Wang et al.  who look5Cucumeropsis mannii seed oil, we recommend further studies into the biological activities of the plant oil and also establish more information on the specific compounds responsible for each biological activity.The results of this study showed that BPA exposure in rats increased serum and adipose tissue levels of cholesterol, triglycerides, LDL‐C, leptin, coronary risk index, atherogenic index, and body weight with decreased HDL‐C, and adiponectin. Interestingly, CMSO treatment reduced the toxicity caused by BPA in rats by modulating the body weight, adiponectin and leptin levels, and lipid profiles in serum and adipose tissue. The findings of this study demonstrate CMSO's therapeutic potential capable of alleviating BPA‐induced dyslipidemia and adipokine dysfunction in male rats. However, as sufficient studies have not been done on The authors declare no conflict of interest.This study received ethical approval number EBSU/BCH/ET/20/003 from the Ethical Committee of the Biochemistry Department, EBSU, and followed their recommendations.All the authors of this manuscript gave their consent to publish.


This study was performed to investigate the relationship between the preoperative cachexia index (CXI) and long‐term outcomes in patients who have undergone radical resection of pancreatic ductal adenocarcinoma (PDAC).In total, 144 patients who underwent pancreatic resection for treatment of PDAC were retrospectively analyzed. The relationship between the CXI and the patients' long‐term outcomes after PDAC resection was investigated. The CXI was calculated based on the preoperative skeletal muscle index, serum albumin level, and neutrophil‐to‐lymphocyte ratio. After propensity‐score matching, we compared clinicopathological features and outcomes.P = 0.0118), R1 resection , and a low CXI  were independent and significant predictors of disease‐free survival (DFS) after PDAC resection. Moreover, a low CXI  was an independent and significant predictor of overall survival (OS) after PDAC resection. After propensity‐score matching, the low CXI group had a significantly worse prognosis than the high CXI group for both DFS and OS.The multivariate analysis showed that lymph node metastasis (hazard ratio [HR], 1.93; 95% confidence interval [CI], 1.16–3.23; The CXI can be a useful prognostic factor for DFS and OS after pancreatic resection for treatment of PDAC. The CXI can be a useful prognostic factor for disease‐free survival and overall survival after pancreatic resection for treatment of PDAC. Cachexia is associated with more than 50% of cancer deaths and causes death in ~30% of patients with cancer.In this study we examined the possibility that the CXI could be a better prognostic factor for PDAC than sarcopenia and the NLR, which have already been reported as prognostic factors for PDAC.22.1This retrospective study was conducted using prospectively collected and maintained data. The study included 144 consecutive patients with cytohistologically proven PDAC who underwent curative‐intent pancreatectomy from January 2014 to December 2021 at the Kyushu Cancer Center. The cases that resulted in nonresection were excluded from the analysis. In this study, we used the 7th edition of the classification of pancreatic carcinoma by the Japan Pancreas Society. When classified by initial resectability, 131 cases were classified as resectable, seven as borderline resectable with artery involvement, and six as borderline resectable with portal vein involvement  × height (m)] × albumin/NLR. The SMI was calculated by measuring the major and minor diameters of the iliopsoas muscle at the third lumbar vertebra using preoperative computed tomography images. The SMI was calculated using the following formula: iliopsoas major axis (mm) × iliopsoas minor axis (mm) × π/100.2.33). The CRP‐to‐albumin ratio was calculated by dividing CRP by serum albumin. The CONUT score was calculated using the serum albumin concentration, peripheral lymphocyte count, and total cholesterol concentration: (a) albumin concentrations of 3.5, 3.0–3.49, 2.5–2.99, and <2.5 g/dL were scored as 0, 2, 4, and 6 points, respectively; (b) total lymphocyte counts of 1600, 1200–1599, 800–1199, and <800 cells/mm3 were scored as 0, 1, 2, and 3 points, respectively; and (c) total cholesterol concentrations of 180, 140–179, 100–139, and <100 mg/dL were scored as 0, 1, 2, and 3 points, respectively. The CONUT score was defined as the sum of (a), (b), and (c).Preoperative blood samples were obtained from all patients within 1 week before surgery. Blood tests in the preoperative treatment group also showed the results immediately before surgery, and the NLR, PLR, and lymphocyte‐to‐monocyte ratio (LMR) were calculated based on the preoperative blood values. The mGPS was calculated as previously described.2.4Pathological evaluation of surgical specimens was based on the tumor‐node‐metastasis (TNM) classification system of malignant tumors by the 7th edition of the classification of pancreatic carcinoma by the Japan Pancreas Society. Evaluation of the excised margin was performed as follows. R0 resection was defined as the absence of tumor cells on the pancreatic resection margin, nerve plexus dissection margin, portal vein dissection surface, posterior dissection surface, and bile duct dissection margin. By contrast, R1 resection was defined as the presence of tumor cells on these margins. In all cases, rapid intraoperative pathological diagnosis of the pancreatic resection margin was conducted, and additional resection was performed when tumor cells were still present on the stump surface.2.5According to the institutional protocols, 49 patients received modern combinatorial chemotherapy (including neoadjuvant therapy with either a 5‐fluorouracil‐based chemotherapy [FOLFIRINOX/FOLFOX] regimen or gemcitabine‐based [gemcitabine/nab‐paclitaxel] regimen or gemcitabine and S‐1 combination therapy) with or without dose modifications as deemed appropriate by the treating oncologists. Chemotherapy was administered on a 2‐ or 4‐week cycle depending on the specific regimen. Total cycles of chemotherapy were counted based on the total number of cycles administered during induction chemotherapy. Two selected patients received chemoradiotherapy as neoadjuvant therapy, and this was determined according to surgical/oncological recommendations in general based on the margin risk. In accordance with the institutional protocol, chemoradiotherapy consisted of photon/proton external beam therapy with a 50.4‐Gy dose delivered in 28 daily fractions over 5 weeks using 3D conformal or intensity‐modulated techniques with concurrent radiosensitizing chemotherapy.2.6Postoperative adjuvant chemotherapy was introduced within 12 weeks after surgery and continued for 6 mo. The chemotherapy regimens included gemcitabine monotherapy, S‐1 monotherapy, gemcitabine and S‐1 combination therapy, or gemcitabine and nab‐paclitaxel combination therapy. No patients received postoperative radiation. After the operation, a blood examination including measurement of tumor markers  and a computed tomography examination was performed every 3 mo for at least 5 y to screen for postoperative recurrence.2.7The background factors of the patients were varied and did not represent a homogeneous population. Statistical methods to investigate the impact of cachexia on such a target patient population requires matching the background factors of the better and worse CXI groups as much as possible. Thus propensity‐score matching analysis was conducted for the patients. The cutoff value of CXI was determined by a receiver‐operating characteristic curve using the survival status at the 3‐y follow‐up in strata of sex. Two factors of maximum tumor size and lymph node metastasis of the metastatic lesion were selected for propensity‐score matching analysis. The caliber for the matching was set as 0.2.2.8U test. Associations between variables were determined by Fisher's exact test or the χ2 test. Multivariate analysis of disease‐free survival (DFS) and overall survival (OS) after pancreatectomy for PDAC was performed using a Cox proportional regression model with a backward elimination stepwise approach. DFS and OS in both the preoperative treatment group and the nonpreoperative treatment group were calculated as years after resection. The survival curve was calculated using the Kaplan–Meier method with the log‐rank test. Statistical analyses were performed using JMP Pro 15.1 . A P‐value of <0.05 was considered statistically significant.The clinicopathological records of all 144 patients were collected and retrospectively reviewed. Differences between the high‐CXI group and low‐CXI group were assessed by the Mann–Whitney 33.1n = 40), delayed gastric emptying (n = 12), bile leakage (n = 1), ascites (n = 7), deep vein thrombosis (n = 11), delirium (n = 9), and wound infection (n = 21).The patients' characteristics are shown in Table 3.2n = 64) and low‐CXI group (n = 80) showed that the body mass index (BMI) and serum albumin concentration were significantly higher in the high‐CXI group than in the low‐CXI group , patients with low CXI (n = 23) had a significantly worse prognosis than those with high CXI , patients with low CXI (n = 34) had a significantly worse prognosis than those with high CXI , high DUPAN‐2 concentration (P = 0.0016), high SPan‐1 concentration (P = 0.0021), tumor size of >2.8 cm (P = 0.0001), lymph node metastasis (P = 0.0001), R1/2 resection (P < 0.0001), and inclusion in the low‐CXI group (P < 0.0001) were significantly associated with DFS after resection of PDAC (Table P = 0.0118), R1/2 resection , and the low‐CXI group  were independently associated with DFS in patients who underwent pancreatectomy for PDAC , tumor size of >2.8 cm (P = 0.0376), lymph node metastasis (P = 0.0081), and inclusion in the low‐CXI group (P < 0.0001) were significantly associated with OS after resection of PDAC (Table P = 0.0002) was independently associated with OS in patients who underwent pancreatectomy for PDAC . Propensity‐score matching analysis was conducted for all 144 PDAC patients; from 144 patients, 108 were selected, among whom 54 patients with high CXI and 54 patients with low CXI. The patients' characteristics and tumor characteristics after propensity‐score matching are shown in Table 4Prognostic factors other than the cancer stage, such as sarcopenia and nutritional predictors, have been studied in recent years. We demonstrated that the CXI, which is calculated using the SMI and NLR, was a prognostic predictor of postoperative outcomes in patients who underwent pancreatic resection for PDAC. This is the first study to show that a low CXI is closely related to poor clinical outcomes in patients with PDAC.Cancer cachexia, a multifactorial syndrome primarily defined by the 2007 Cachexia Consensus Conference as a complex metabolic syndrome associated with underlying illness and characterized by loss of muscle with or without loss of fat mass, is associated with ~30% of cancer‐related deaths.The clinical characteristics of cancer cachexia are a poor nutritional status, systemic inflammation, and loss of muscle mass. Malnutrition is a frequent and serious problem in patients with various cancers, and it negatively affects many patients' quality of life and prognosis.P = 0.8904). Therefore, we think that it is good to measure CXI before neoadjuvant treatment. If CXI measured before neoadjuvant treatment is low, rehabilitation and nutritional interventions may increase CXI before surgery. This may lead to an improved prognosis. However, there were no cases in which CXI was frequently measured before and after surgery in this study. We think that it is necessary to consider it in the future.Compared with the high‐CXI group in this study, the low‐CXI group had more advanced‐age patients with a lower BMI, albumin concentration, LMR, total lymphocyte count, and PNI and a higher NLR, PLR, mGPS, and CONUT score Table . Hence, In the multivariate analysis, patients with a low CXI showed a poor prognosis with respect to both DFS and OS Tables , 3. As aA number of reports have proposed that the CXI is a novel index for estimating cachexia that also correlates with the prognosis of malignant tumors such as colorectal liver metastasis, diffuse large B‐cell lymphoma, non‐Hodgkin's lymphoma, and advanced non‐small cell lung cancer (NSCLC).A study is underway in Japan to improve cancer cachexia with the use of anamorelin, a ghrelin receptor agonist that can be administered orally.Our study had two main limitations. First, it was a single‐center retrospective study involving a relatively small number of patients. Second, the retrospective nature of the study prevented comparison of blood data and imaging findings because of differences in the number of chemotherapy courses and the contents of chemotherapy. Therefore, a large‐scale prospective study is needed to validate the results.The preoperative CXI was a reliable indicator to assess the DFS and OS after pancreatectomy for treatment of PDAC. The use of this index for clinical decision‐making will facilitate the development of a more accurate risk stratification system combined with tumor staging of PDAC. In conclusion, the CXI seems to be a useful predictor of poor DFS and OS after pancreatectomy for PDAC.T.S. participated in writing the article. T.S. and K.S. participated in the conception and design of the study. Y.M., E.O., and T.I. participated in the acquisition of data. Y.N., M.S., M.Y., and M.M. participated in the statistical analysis and interpretation of data. K.S. participated in reviewing the article. Y.T. participated in reviewing the article and providing final approval of the version to be submitted.No funding was received for this study.All authors declare no conflicts of interest for this article.This study protocol complied with the ethical guidelines of human clinical research established by the Japanese Ministry of Health, Labour and Welfare, as well as with the 1964 Helsinki Declaration and its later amendments. The study was approved by the Ethics and Indications Committee of the National Hospital Organization Kyushu Cancer Center (No. 2019–54). Informed consent was obtained from all individual participants included in the study.Figure S1.Click here for additional data file.


Malignant hemangioendothelioma is an endothelial cancer with heterogeneous clinical behavior that can range from indolent to aggressive, of which the majority are epithelioid (EHE). Its incidence and demographics have not been previously well defined in a large cohort.n = 1986). Survival analyses were performed on a subset of patients within the SEER‐18 database with survival information available (n = 417). Outcomes included incidence, demographics of patients newly diagnosed with hemangioendothelioma, extent of disease at presentation, and overall survival.This retrospective analysis used the US Cancer Statistics National Program of Cancer Registries – Surveillance Epidemiology End Results (SEER) combined database to identify patients in the US newly diagnosed with hemangioendothelioma between the years of 2001 and 2017 , incidence rates were stable. Skin and connective tissues were the most common presenting sites (33.4%), followed by liver (24.5%), lung (17.6%), and bone (12.5%). Median age at diagnosis was 55 years; 27.2% of patients were pediatric, adolescent, or young adult (<40 years). At presentation, 36.4% of patients had localized disease; 21.6% presented with regional and 41.7% with distant metastases. Observed survival at 3 years was 79.7%, 70.7%, and 46.0% for patients presenting with local, regional, and distant disease and most deaths occurred within the first 2 years.Malignant hemangioendothelioma is ultra‐rare but meaningfully impacts affected patients. These data may provide benchmarks for comparison of new approaches to hemangioendothelioma therapy and highlight poor survival outcomes. Hemangioendothelioma is ultra‐rare with incidence of 0.4 cases per million person‐years but meaningfully impacts affected patients. These data may provide benchmarks for comparison of new approaches to hemangioendothelioma therapy and highlight poor survival outcomes. The overall incidence has previously been reported as 0.38 per 1,000,000 person‐years in a large European database,Due to the extreme rarity of EHE, the patterns of initial presentation have not been well characterized in a large cohort. A consensus statement of EHE experts emphasized the need for data to guide surveillance strategies and to establish clinical benchmarks for future clinical trials.22.1malignant hemangioendotheliomas, subtypes such as retiform haemangioendothelioma (9136/1), papillary intralymphatic angioendothelioma (9135/1), composite haemangioendothelioma (9136/1), pseudomyogenic haemangioendothelioma (9138/1) and kaposiform haemangioendothelioma have “benign” ICD‐O‐3 codes and are thus not included in national registry data. For cases diagnosed from 2003 to 2017, 100% of the population is covered for all 50 U.S. states and the District of Columbia. In 2001 and 2002, cases that were diagnosed in Mississippi are not available, so 99% of the U.S. population is covered. In keeping with privacy requirements, case numbers <16 cases are suppressed. Survival data were available on the subset of cases both contained within SEER 18 databases and who had hemangioendothelioma diagnosed as their first malignancy.IRB approval was obtained (Providence 202000564) and this research was conducted in accordance with Helsinki principles. De‐identified aggregate national registry data was extracted from public use databases of the US Cancer Statistics(USCS) National Program of Cancer Registries – Surveillance Epidemiology End Results (SEER) submission for the years 2001–2017.2.2p‐value for significance was set a priori at 0.05. For incidence rate calculations, data were age‐adjusted to the 2000 US standard population . Confidence interval (CI) estimation was performed using the methods of Tiwari et al.2.3Data were extracted using SEER*Stat software version 8.4.0.1 released May 16, 2022.33.1Between 2001 and 2017, a total of 1986 cases of hemangioendothelioma were reported in the US . New cases increased from 100 in the year 2001 to 122 in 2017  and 9133/3 (EHE) were similar; therefore, the populations were kept together for primary analysis. The most common sites of presentation were in the connective tissues/subcutaneous tissues/skin  while 83.3% were white, 10.5% Black, 3.9% Asian, and the remaining persons were unknown/unspecified, or in a racial group containing fewer than 16 cases Figure . 11.0% o3.4Local‐regional‐distant (LRD) staging information was available for 1618 of the 1986 patients (81.5%). 41.7% presented with distant metastatic disease while 21.6% of patients presented with regional disease, including 269 patients by direct extension alone, 42 patients by regional lymph nodes alone, and 39 patients regional by both modes. Approximately one‐third of patients (36.7%) presented with localized disease Figure .3.5n = 247; survival significantly worsened for distant presentation). Patients with primary presentation in soft tissue/skin/subcutaneous had significantly improved outcomes  versus those presenting with lung/mediastinal disease ; outcomes for liver  or bone  were intermediate.Survival information was available for a subset of 417 out of 1986 patients captured within SEER‐18 database Section . Observe4Hemangioendothelioma is an ultra‐rare sarcoma with a paucity of data to guide optimal treatment and counseling of patients. To our knowledge this is the largest cohort of malignant hemangioendothelioma/EHE patients reported to date. The US incidence rate in this series (120 new cases diagnosed in the US per year) is similar to that recently reported in France and approximately double the previously reported US incidence.Although EHE is often considered an indolent cancer, we observed 3‐year OS of only 63.5% which was far worse than expected survival based on actuarial data matched on sociodemographic and socioeconomic factors. This, combined with the fact that most deaths occurred within 5 years from diagnosis and most of those within the first 2 years, suggests that EHE is the major factor driving mortality and further supports the use of 1‐ or 3‐year OS with comparison to historical data as a feasible and clinically appropriate clinical trial endpoint. As expected, outcomes in patients with advanced disease at presentation were especially poor. This is consistent with prior reports identifying pulmonary infiltrates, pleural effusion, and ascites as negative prognostic signs in EHE.WWTR1‐CAMTA1 gene fusions in patients with hemangioendothelioma and the consistency in demographics and survival outcomes that we demonstrated between the separate EHE and malignant hemangioendothelioma groups, we believe that this data still reflects a useful comparator.A limitation of our data set is that considering the rare nature of this tumor, misclassification bias is possible and genomic or biomarker data to confirm the diagnosis were not available in this national population‐based registry. Other types of hemangioendothelioma such as those with other gene fusions are likely also included in the “hemangioendothelioma” group. A second limitation is that the USCS registry only includes malignant tumors. Several types of hemangioendothelioma, including those with aggressive potential such as kaposiform hemangioendothelioma, have diagnosis codes which are categorized as benign in ICD‐O‐3 categorization of malignancies. These types  were thus not recorded in USCS or possible to include in our analyses. Although there have been improvements in survival of patients with Kaposiform hemangioendothelioma in recent years due to sirolimus based regimens, we do not expect that would impact our findings.As noted in a recent EHE consensus paper,Kelly G. Paulson: Conceptualization ; data curation ; formal analysis ; investigation ; methodology ; project administration ; writing – original draft ; writing – review and editing . Vinod Ravi: Writing – review and editing . Brian P. Rubin: Writing – review and editing . Min Park: Writing – review and editing . Elizabeth T. Loggers: Writing – review and editing . Lee D. Cranmer: Writing – review and editing . Michael J. Wagner: Conceptualization ; formal analysis ; investigation ; methodology ; project administration ; writing – original draft ; writing – review and editing .Work supported in part through the Paul G. Allen Research Center at Swedish Cancer Institute (K.G.P.) and by support to the Sarcoma Oncology Program of the University of Washington from Curt and Elizabeth Anderson and P30CA015704 . Dr. Cranmer was supported in part by the Curt and Elizabeth Anderson Endowed Professorship in Sarcoma Research, and by funding from Steve and Jane Urner.KGP, VR, MP, BPR—nothing to disclose. ETL—Clinical trial funding from BioAtla, Ayala, SpringWorks. LDC—Clinical trial funding from Eli Lilly, AADi, BluePrint Medicine, Iterion, Gradalis, Philogen, Advenchen Laboratories, and CBA Pharma. Honoraria or has served on advisory boards for Daaichi Sankyo, BluePrint Medicines and Regeneron. MJW—Clinical trial support from Deciphera, Adaptimmune, GSK, Athenex, Foghorn Therapeutics, Shaqsi, Presage Biosciences, Inhibrx, Incyte. Consulting fees from Adaptimmune, Epizyme, Aadi, Deciphera.Figure S1:Click here for additional data file.


EGFR mutations remains unclear.Vascular endothelial growth factor (VEGF) has been identified as one of the resistant mechanisms to epidermal growth factor receptor‐tyrosine kinase inhibitors (EGFR‐TKIs). However, the relationship between the efficacy of osimertinib and protein expression of VEGF family members in patients with advanced non‐small cell lung cancer (NSCLC) harboring EGFR major mutations (del19 or L858R) receiving first‐line osimertinib were eligible as the osimertinib (Osi) group, whereas 43 patients receiving first‐ or second‐generation EGFR‐TKIs were compared with the control group. The expression of vascular endothelial growth factor receptor 2 (VEGFR2) and vascular endothelial growth factor C (VEGF‐C) in the tumor specimens was analyzed using immunohistochemistry.A total of 76 patients with advanced NSCLC with VEGFR2 and VEGF‐C were highly expressed in 65.8% and 51.3% of patients, respectively, in the Osi group, and 69.7% and 76.7%, respectively, in the control group. High VEGFR2 and VEGF‐C levels were significantly associated with poor performance status (PS) and female sex, respectively. In the Osi group, patients with co‐high expression of VEGFR2 and VEGF‐C showed significantly worse progression‐free survival (PFS) and overall survival (OS) than those without co‐high expression. In del19, VEGFR2 was a significant predictor of PFS and OS and independent predictor of OS in multivariate analysis. In L858R, co‐high expression of VEGFR2 and VEGF‐C was identified as a significant predictor of PFS and OS and independent predictor of PFS.EGFR‐mutant NSCLC cells. Increased expression of VEGFR2 was identified as a significant prognostic factor in patients with EGFR del19 mutation who received osimertinib, whereas co‐high expression of VEGFR2 and VEGF‐C was a significant predictor for those with EGFR L858R mutation.VEGFR2 and VEGF‐C are highly expressed in  EGFR‐mutant NSCLC. Co‐high expression of VEGFR2 and VEGF‐C are related to worse prognosis for patients receiving osimertinib. High VEGFR2 expression was identified as a significant predictor in EGFR del19 mutation. Co‐high expression of VEGFR2 and VEGF‐C was a significant predictor for EGFR L858R mutation.VEGF‐agonist and ligand are highly expressed in  EGFR) mutations are major oncogenic alterations in non‐small cell lung cancer (NSCLC), particularly adenocarcinoma, and have been identified as molecular targets.EGFR tyrosine kinase inhibitors (TKIs) exist including first, second, and third generations available for EGFR‐mutant NSCLC.Epidermal growth factor receptor  increased VEGF production.EGFR cells and that the EGFR signaling pathway can induce VEGFR2 pathway activation, suggesting the enhancement of antitumor effects of EGFR‐TKI through VEGFR2 inhibition.7 A recent clinical trial demonstrated that ramucirumab (a VEGFR2 inhibitor) plus erlotinib achieved superior progression‐free survival (PFS) compared with erlotinib alone in untreated EGFR‐mutated NSCLC.EGFR mutated NSCLC remains unknown, a phase 1/2 trial revealed that the combination of osimertinib and bevacizumab met the primary endpoint of progression‐free survival at 12 months; thus, a randomized phase 3 trial comparing osimertinib plus bevacizumab and osimertinib is planned.EGFR‐mutated NSCLC.EGFR‐mutated NSCLC. In particular, the efficacy of osimertinib and the expression of VEGF in tumor specimens have not been described. Our previous investigations confirmed that VEGF‐A, ‐B, and ‐D as VEGF‐ligand and VEGFR1 and 3 as VEGFR‐receptor were not suitable for immunohistochemistry in human tumor specimens because of nonspecific staining.Recently, it has been reported that the expression of vascular endothelial growth factor receptor 2 (VEGFR2) is higher in EGFR‐TKI‐resistant cell lines than in TKI‐sensitive cell lines.EGFR‐mutated NSCLC, using Ki‐67 as a tumor cell proliferation marker and system alanine–serine–cysteine amino acid transporter‐2 (ASCT2) as amino acid transporter.Based on this background, we conducted a clinicopathological study to evaluate the prognostic impact of VEGF‐ligand and VEGFR‐receptor expression after osimertinib treatment in patients with advanced or metastatic EGFR mutations were treated with osimertinib as a first‐line treatment in our institution. The inclusion criteria were a therapeutic history of first‐line osimertinib, major EGFR mutant tumors with deletions in exon 19 (del19) or exon 21 codon p.Leu858Arg (L858R), and sufficient tumor specimens for immunohistochemistry. Of these, six patients exhibited EGFR mutations  without del19 or L858R, and 23 patients did not have sufficient tumor specimens for immunohistochemistry before osimertinib therapy. Therefore, 76 patients  were eligible for this study (Osi group). Compared with the 76 patients receiving osimertinib, conversely, the patients with advanced NSCLC harboring major EGFR mutations (del19 or L858R) who received first‐ or second‐generation EGFR‐TKI were also analyzed as a control group. Forty‐three patients were eligible for further analysis between September 2011 and November 2018. 2. Most patients in the control group were definitively diagnosed by cytological examination; thus, recruiting more patients was difficult owing to the lack of tumor specimens. EGFR mutation testing using Cobas was performed according to the manufacturer's protocol.Between August 2018 and July 2021, 105 patients with advanced or metastatic NSCLC harboring Clinical data, such as age, sex, performance status (PS), smoking history, radiological examination, and survival information, were extracted from the medical records. This study was approved by the Institutional Ethics Committee of the International Medical Center of Saitama Medical University , Hidaka City, Japan. The Ethical Committee waived the need to obtain written informed consent for the use of human tissues to participate from the patients owing to the retrospective nature of the study.Osimertinib (80 mg/day) was orally administered as first‐line treatment in 76 patients. In the control group, gefitinib 250 mg/day, erlotinib 150 mg/day, or afatinib 30 mg/day as first‐ or second‐generation EGFR‐TKI was orally administered. Physical examinations were performed and complete blood counts, biochemical tests for liver and renal dysfunction, electrolytes, and adverse events were examined by the chief physician. Toxicity was graded based on the Common Terminology Criteria for Adverse Events, version 4.0.Immunohistochemical staining was performed as previously described.Sections were evaluated by at least two researchers (KK and HI) using a light microscope (×200 and ×400 magnification) in a blinded fashion. In the case of discrepancies, both investigators evaluated the slides simultaneously until a final consensus was reached. The investigators were blinded to the patient outcomes.t (unpaired t) and χ2 tests were used for continuous and categorical variables, respectively. Statistical significance was set at p < 0.05. PFS was defined as the time from initial EGFR‐TKI treatment to disease progression or death. Overall survival (OS) was defined as the time from the initial EGFR‐TKI treatment to death from any cause. The Kaplan–Meier method was used to estimate survival as a function of time, and survival differences were analyzed using the log‐rank test. Univariate and multivariate analyses were performed using logistic regression. All statistical analyses were performed using GraphPad Prism  and JMP 14.0 .Student's EGFR mutation status, del19 and L858R were detected in 39 (51.3%) and 37 (48.7%) patients, respectively.Patient characteristics based on VEGFR2 and VEGF‐C expression are listed in Table male = 18, nfemale = 25; age, median = 70 years, range = 38–86 years) in the control group are shown in Table The demographics of the 43 patients , and those of high VEGF‐C expression in the Osi and control groups were 51.3% (39/76) and 76.7% (33/43), respectively (p = 0.006). In the Osi group, the incidences of scores 1, 2, 3, and 4 were 0 (0.0%), 5 (6.6%), 21 (27.6%), and 50 (65.8%), respectively, for VEGFR2, and 4 (5.3%), 10 (13.1%), 23 (30.3%), and 39 (51.3%), respectively, for VEGF‐C. The patients with no expression for VEGFR2 and VEGF‐C were not observed. High VEGFR2 expression was significantly related to poor PS, and high VEGF‐C expression was closely associated with elderly age  and 17 (22.4%) patients, respectively, for the Osi group and 26 (60.5%) and 17 (39.5%) patients, respectively, for the control group. Representative images of VEGFR2 and VEGF‐C expression are shown in Figure Table Moreover, the ORR and DCR between del19 and L858R were not significantly different based on VEGFR2 and VEGF‐C expression.In the Osi group, the median PFS and OS were 566 and 826 days, respectively. Fifty‐one patients experienced tumor recurrence, and 43 died due to progressive disease. In the control group, the median PFS and OS were 531 and 1082 days, respectively. A total of 37 patients experienced tumor recurrence, and 28 died because of progressive disease. Kaplan–Meier survival curves based on VEGFR2 and VEGF‐C expression were constructed in the Osi and control groups Figure  and betwEGFR mutation status (del19 and L858R) was performed in both groups. In the Osi group, the PFS and OS based on del19 EGFR mutation were significantly worse in the patients with high VEGFR2 expression than in those with low VEGFR2 expression , 26 (34.2%) patients were treated with sequential therapy after failure of osimertinib. Of these 26 patients, 15 patients received chemoimmunotherapy such as carboplatin, paclitaxel, bevacizumab and atezolizumab, six patients were treated with platinum‐based chemotherapy such as carboplatin plus pemetrexed or carboplatin plus nab‐paclitaxel, and five patients underwent first‐ or second‐generation EGFR‐TKIs such as gefitinib or afatinib. In the control group (n = 43), on the other hand, 22 (51.2%) patients were treated with sequential therapy. Of these 22 patients, seven received carboplatin plus pemetrexed, 12 patients were treated with osimertinib for the detection of T790M, and three patients underwent pemetrexed alone.In the Osi group .Kyoichi Kaira has received research grants and a speaker honorarium from Ono Pharmaceutical Company, Boehringer Ingelheim, Chugai Pharmaceutical, Taiho Pharmaceutical, Eli Lilly Japan and AstraZeneca. Atsuto Mouri and Ou Yamaguchi received a speaker honorarium from Eli Lilly, Taiho Pharmaceutical, Pfizer, Chugai Pharmaceutical and AstraZeneca. Hiroshi Kagamu has received research grants and a speaker honorarium from Ono Pharmaceutical Company, Bristol‐Myers Company, Boehringer Ingelheim, MSD, Daiichi Sankyo Company, Chugai Pharmaceutical, Taiho Pharmaceutical, Merck Biopharma Company, Eli Lilly Japan and AstraZeneca. Hisao Imai has received research grants and a speaker honorarium from Ono Pharmaceutical Company, AstraZeneca and Bristol‐Myers Company.The present study was approved by the Institutional Ethics Committee of the International Medical Center of Saitama Medical University, Hidaka city, Japan. The Ethical Committee waived the need to obtain written informed consent for the use of human tissues to participate from the patients owing to the retrospective nature of the study.Table A1. Patient demographics based on VEGFR2 and VEGF‐C expression (Control group).Click here for additional data file.Table A2. Comparison of patient demographics between Osi‐group and Control group.Click here for additional data file.Table A3. Efficacy according to VEGFR2 and VEGF‐C expression.Click here for additional data file.Table A4. Univariate and multivariate analysis in patients with del19 (n = 39).Click here for additional data file.Table A5. Univariate and multivariate analysis in patients with L858R (n = 27).Click here for additional data file.


These results show altered cellular and molecular changes due to reduced estrogen levels. Developing new treatments for hypoestrogenic vaginal symptoms rely on better understanding of not only the cellular changes, but also the altered vaginal ECM environment. Further studies using this mouse model has the potential to advance women's vaginal health treatments and aid in understanding the interplay between organ systems in both healthy, aged, and diseased states.Loss of estrogen as a result of aging, pelvic cancer therapy, genetics, or eating disorders affects numerous body systems including the reproductive tract. Specifically, a chronic hypoestrogenic state fosters debilitating vaginal symptoms like atrophy, dryness, and dyspareunia. Current treatment options, including vaginal estrogen and hyaluronan (HA), anecdotally improve symptoms, but rectifying mechanisms are largely understudied. In order to study the hypoestrogenic vaginal environment, in particular the extracellular matrix (ECM), as well as understand the mechanisms behind current treatments and develop new therapies, we characterized a reliable and reproducible animal model. Bilateral ovariectomies (OVX) were performed on 9‐week‐old CD1 mice. After 1 month of estrogen loss due to ovarian removal, a phenotype that is similar to human vaginal tissue in an estrogen reduced state was noted in mice compared to sham‐operated controls. The uterine to body weight ratio decreased by 80% and vaginal epithelium was significantly thinner in OVX compared to sham mice. Estrogen signaling was altered in OVX, but submucosal ERα localization did not reach statistical differences. HA localization in the submucosal area was altered and CD44 expression decreased in OVX mice. Collagen turn‐over was altered following OVX. The inflammation profile was also disrupted, and submucosal vaginal CD45 Murine ovariectomy resulted in decreased circulating estrogen as evidence by smaller uterine: BW ratio. The hypoestrogenic vaginal state displayed altered estrogen signaling, hyaluron localization, collagen turn over, and inflammatory profile. Therefore, vaginal HA has become an alternative, nonhormonal treatment for genitourinary menopausal symptoms. It alleviates vaginal dryness and dyspareunia by retaining water molecules, increasing tissue hydration, and inducing physiologic changes which reverse many vaginal issues related to loss of estrogen  Center for Comparative Medicine. They were socially housed, exposed to 12 h light/12 h dark cycles, and given free access to standard food and water. All procedures described herein were approved by BCM Institution for Animal Care and Use Committee, AN‐7475, and completed in compliance with all recommendations in the Guide for the Care and Use of Labratory Animals of the National Institute of Health.2.2n = 4–5 per group): sham and OVX. Mice were anesthetized using 0.5 L/min isoflurane. Bilateral, 0.5 cm incisions were created in the dorsal flank. In Sham control mice, the ovaries were visualized, then the incisions were closed with 5‐0 vicryl and/or Dermabond skin glue. In OVX mice, once the ovaries were visualized, the ovarian ligament was clamped and cauterized, and each ovary was excised. The incisions were again closed with 5‐0 vicryl and/or Dermabond skin glue. Sham and OVX mice received standard postoperative care including observation and administration of analgesics for 3 days following the procedure.Mice were randomly separated into two groups . Sections were cut to 5 μm, rehydrated, and stained using standard hematoxylin and eosin.To determine epithelial thickness, 20X images were taken using Leica DMi8 microscope with a Leica DFC4500 camera and Leica Application Suite X (LAS‐X). ImageJ software was used to measure the epithelial thickness along 10 points of each image, 3 images per mouse were analyzed and all measurements were averaged.n = 3 per mouse) images were taken using Leica DMi8 microscope with a Leica DFC4500 camera and LAS‐X. ImageJ was used to determine the percent positive area above a set threshold which was kept constant for each image and reported as percent positive area .HA staining was evaluated using HA binding protein (HABP) and a protocol adapted from Cleveland Clinic's Programs of Excellence in Glycosciences. Briefly, FFPE tissue sections were rehydrated in a series of graded ethanol. Slides were equilibrized in PBS, endogenous avidin and biotin were blocked . Slides were incubated in normal goat serum (150 μL/10 mL) followed by biotinylated HABP  for 1 h. After washing, the signal was amplified using avidin/biotin complex  and detected using DAB . Slides were counterstained using hematoxylin . 20X (n = 3 per mouse) images, under both bright field and circular polarized light, were taken using Leica DMi8 microscope with a Leica DFC4500 camera and LAS‐X. ImageJ was used to determine total submucosal area. Using the polarized light images, color masks were created to measure positive red area and positive yellow/green area as previously described  staining. Slides were stained in PSR solution  for 1 h following deparaffinization to water. After two rinses with acidified water , samples were dehydrated with three changes of 100% ethanol. Finally, slides were cleared with xylene and cover slipped for subsequent imaging. 20X (n = 3 per mouse) images were taken using Leica DMi8 microscope with a Leica DFC4500 camera and LAS‐X. ImageJ was used to determine submucosal area and positive cell number to calculate number of cells per submucosal area.Immunohistochemistry (IHC) was used to localize ERα, HA receptor (CD44), and inflammation markers  as described previously  according to the manufacturer's instructions, including DNA elimination step.2 . Samples were performed in duplicate using Power SYBR Green PCR Master Mix  and Bio‐Rad CFX384 thermocycler. Primer sequences used are listed in Table 2.5t‐test with 95% confidence level were used to determine significance (n = 4–5/group). Post hoc power analysis was conducted after the study completion to determine that at least 80% power of detecting any difference in each measured outcome at Type I error α = 0.05 was achieved using two‐sided, two‐sample t‐test for mean differences which yielded N = 6 as total sample size for two groups . p < 0.05 was considered significant and specific p values are noted in graphs.Graphpad Prism Version 8 was used to graph and analyze data. All data are presented as individual data points on graphs and include the mean ± SD. Two tailed 33.1Understanding how estrogen, produced primarily in the ovary, can affect intact vaginal tissue is vital to understanding how estrogen can affect vaginal healing. Here, murine bilateral OVX was used to model a vaginal hypoestrogenic state. Both ovaries were removed from 9‐week‐old CD1 female mice with sham‐operated mice as controls Figure . The ove3.22 . In our study, Fos expression is downregulated 3.4X in the OVX mice compared to sham mice , but a decreased Has3 expression (p = 0.01) was found when comparing OVX mice to sham mice  and significantly fewer small fibers (p = 0.008) in the OVX mice compared to sham mice , which plays a role in the regulation of the ECM. Generally, many of the downregulated ECM‐related genes in OVX correlate to the phenomena of vaginal tissue exhibiting overall reduced tissue strength after loss of circulating estrogen   and non‐GAG‐related components such as collagens I and III and elastin  was decreased and altered large to small collagen fiber ratio was noted. Perhaps this increased tendency for large, Type I collagen in the OVX condition can contribute to the increased tissue stiffness often associated with menopausal vaginal tissue , and HA and collagen turn over in other vaginal injury models such as surgical injury, radiation therapy, and fibrosis.5Developing new treatments for hypoestrogenic vaginal symptoms rely on better understanding the altered cellular and tissue environment. The bilateral OVX mouse model recapitulates the thinner epithelium, altered HA localization and signaling, and dysregulated collagen content observed in human postmenopausal vaginal tissue as well as vaginal tissue impacted from premature ovarian failure. Further, ovarian removal results in decreased basal vaginal inflammation, and we propose this could lead to lower defense mechanisms that often make postmenopausal women victims of vaginal infections. Further studies using this model will evaluate the impacts of prolonged estrogen depletion along with investigations into uncovering other mechanisms of ECM dysregulation via biomechanical perturbations. Additionally, this model can be used in other aspects of gynecologic research to establish connections between hormone signaling and wound healing, infection, and/or fibrosis. In summary, the characterization into the submucosal changes of this mouse model, specifically CD4+ cells, HA, and collagen turnover, will pave way for advancement in women's vaginal health discoveries, innovations, and treatment paradigms.Jennifer M McCracken: Conceptualization, Methodology, Validation, Formal Analysis, Investigation, Data Curation, Writing–Original Draft, Writing–Review and Editing, Visualization, Project Administration. Gisele A Calderon: Conceptualization, Methodology, Validation, Formal Analysis, Investigation, Data Curation, Writing–Original Draft, Writing–Review and Editing, Visualization, Project Administration. Quynh N Le: Data Analysis, Writing–Review and Editing. Natasha M Faruqui: Data Curation and Analysis. Swathi Balaji: Data assessment, critical review and editing of the manuscript. Julie CE Hakim: Conceptualization, Supervision, Project Administration, Funding Acquisition.This work was graciously supported by NIGMS 1K08GM135638–01.The authors have no conflict of interest to declare.


AL patients develop the unique toxicities of fluid retention and non‐cardiogenic pulmonary edema during the course of stem cell mobilization. We propose mobilization with CART as effective and safe treatment for AL patients with refractory anasarca.We describe a 63‐year‐old male with systemic immunoglobulin light chain (AL) amyloidosis with cardiac, renal, and liver involvement. After four courses of CyBorD, mobilization with G‐CSF at 10 μg/kg was initiated and CART was simultaneously performed for fluid retention. No adverse events were observed during collection or reinfusion. Anasarca gradually disappeared and he underwent autologous hematopoietic stem cell transplantation. The complete remission of AL amyloidosis has been maintained, and the condition of the patient has remained stable for 7 years. We propose mobilization with CART as an effective and safe treatment option for AL patients with refractory anasarca. A 63‐year‐old male with systemic AL amyloidosis with cardiac, renal, and liver involvement. Computed tomography (CT) scan showing massive ascites at presentation . After 4 courses of CyBorD and mobilization with G‐CSF at 10 μg/kg was initiated. CT scan taken after G‐CSF for mobilization showing an increase in massive ascites . CART was simultaneously performed for fluid retention. CT scan taken after CART and autologous stem cell transplantation showing improvements . Although AL amyloidosis may involve any organ, the heart and kidneys are the most commonly affected organs. The presence and severity of cardiac disease is typically a powerful predictor of overall survival (OS).However, AL patients develop fluid retention and non‐cardiogenic pulmonary edema during the course of stem cell mobilization, which may lead to a mortality rate of 2%–3% in the prechemotherapy phase.We herein presented an AL patient with cardiac involvement who developed fluid retention during the course of stem cell mobilization that was controlled by CART. After autologous HSCT, the patient maintained complete remission (CR) for 7 years. We propose mobilization with CART as an effective and safe treatment option for patients with systemic AL and refractory ascites.29/L, red blood cell count of 360 × 1010/L, hemoglobin concentration of 12.1 g/dL, and platelet count of 22.8 × 109/L. His serum albumin level was 1.48 g/dL , alkaline phosphatase was 787 U/L , β2 microglobulin was 3.1 μg/mL , and estimated urinary total protein was 894 mg/dL . The patient was diagnosed with nephrotic syndrome and kidney biopsy was performed. Light microscopy indicated mesangial expansion with eosinophilic material . A bone marrow examination revealed 3% plasma cells. In laboratory tests, the level of N‐terminal brain natriuretic peptide (NT‐proBNP) was high  and that of troponin I was slightly elevated . A sinus rhythm with a low QRS voltage was observed on electrocardiography (ECG)  Figure . Echocar) Figure .2 on days 1, 4, 8, and 11 every 3 weeks, s.c.), and dexamethasone  and 9–12 (cycles 1 and 2), p.o.) (CyBorD). After four courses of CyBorD, the patient achieved a hematologic complete response. Mobilization with G‐CSF at 10 μg/kg was initiated and fluid retention was detected , followed by the infusion of CD34 at a total dose of 1.9 × 106/kg. His serum albumin level gradually increased to 3.3 g/dL, and the urinary protein creatinine ratio decreased from 4.38 to 1.23. His ALP level normalized (320 U/mL), and NT‐proBNP and troponin I levels both decreased . ECG also normalized , bortezomib (1.3 mg/md Figure . CART wad Figure . The cond Figure .3Cardiac involvement is a major factor affecting the survival of patients with AL amyloidosis. Untreated patients with cardiac amyloidosis have historically had a median survival of 6–12 months; however, the clinical use of molecularly targeted drugs, such as proteasome inhibitors and immunomodulatory agents, monoclonal antibodies such as daratumumab, have markedly prolonged OS.Many patients with AL already have severe organ damage and, thus, supportive care forms the foundation of the success of HSCT in AL. AL patients develop the unique toxicities of fluid retention and non‐cardiogenic pulmonary edema during the course of stem cell mobilization that may lead to a mortality rate of 2%–3% in the prechemotherapy phase.CART, a treatment system for ascites that was developed in 1977, removes unnecessary components in ascites using a filtration membrane and intravenously reinfuses collected proteins. CART is expected to improve quality of life and increase urine output by maintaining hemodynamics through increases in serum albumin levels. CART is safe and effective as a treatment option for large‐volume paracentesis with the infusion of albumin for cancer or cirrhosis.In the present case, fluid retention during the course of stem cell mobilization was controlled by CART and CR was achieved after PBSCT. After 7 years, the patient has remained in CR with the resolution of peripheral edema and a decrease in the level of NT‐proBNP. We propose mobilization with CART as an effective and safe treatment option for AL patients with refractory anasarca.In conclusion, the early diagnosis of AL is important for the prevention of advanced damage and relapse while controlling plasma dyscrasia with highly effective treatments. The decision to treat this condition using aggressive measures with CART must not be delayed until advanced damage.Satoko Oka: Writing – original draft. Kazuo Ono: Writing – original draft.All authors have no conflict of interest.Written informed consent was obtained from the patient to publish this report in accordance with the journal's patient consent policy.


The outbreak of coronavirus disease 2019 (COVID‐19) caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) has posed a great threat to human health. Some severe COVID‐19 patients still carried detectable levels of SARS‐CoV‐2 even after prolonged intensive care unit treatment. However, the immunological features of these COVID‐19 patients with delayed virus clearance (CDVC) are still unclear.n = 9) and recovered (n = 52) cases. The expression of the exhaustion marker PD‐1 on circulating T cells of these patients was measured by flow cytometry.We retrospectively reviewed the clinical and immunological data of 13 CDVC cases, who were admitted into one hospital in Wuhan from February to April 2020. These data were also compared to those of perished (9/L), CD4+ T cells (0.35 × 109/L), and CD8+ T cells (0.28 × 109/L) below their corresponding lower limits of normal range. Similar to the perished group, CDVC cases have higher percentages of CD25+Foxp3+ regulatory T cells (Treg) in circulation. Moreover, enhanced expression of the exhaustion marker PD‐1 on CCR7−CD45RA+ effector, CCR7+CD45RA− central memory, and CCR7−CD45RA− effector memory CD4+ and CD8+ T cells were also observed in CDVC cases.High levels of serum interleukin‐6 (IL‐6), IL‐1β, IL‐8, as well as other inflammatory mediators, were seen in CDVC cases. Severe lymphopenia was observed in CDVC patients with the counts of total lymphocytes (0.9 × 10CDVC patients still have SARS‐CoV‐2 and these cases manifest with severe clinical symptoms due to persistent inflammation. Augmentation of the frequency of circulating Treg, severe lymphopenia, and functional exhaustion of T cells might lead to inefficient clearance of SARS‐CoV‐2. Therefore, enhancing lymphocyte counts and reversing T‐cell exhaustion might be key methods to boost immune responses and eliminate SARS‐CoV‐2 in CDVC patients. CDVC patients still have SARS‐CoV‐2 and these cases manifest with severe clinical symptoms due to persistent inflammation. Augmentation of the frequency of circulating Treg, severe lymphopenia, and functional exhaustion of T cells might lead to inefficient clearance of SARS‐CoV‐2. Therefore, enhancing lymphocyte counts and reversing T‐cell exhaustion might be key methods to boost immune responses and eliminate SARS‐CoV‐2 in CDVC patients. Later, the causative agent of this severe pneumonia was identified as a novel coronavirus and named as 2019 novel coronavirus (2019‐nCoV), and subsequently officially reclassified as the novel severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), while the clinical condition is referred to as coronavirus disease 2019 (COVID‐19) by the World Health Organization (WHO).The typical symptoms of COVID‐19 usually appear 2–14 days after viral exposure. Fever, cough, shortness of breath, and pneumonia are very common in mild cases, whereas severe cases show respiratory, hepatic, gastrointestinal, and neurological complications.n = 9), CDVC (n = 13), and recovered (n = 52) based on outcomes of treatment. Our findings may extend the knowledge and understanding of the factors associated with CDVC.In this study, we performed a comprehensive evaluation of clinical and immunological data of 74 patients admitted to Huoshenshan Hospital, Wuhan. These patients were categorized into three groups including perished  published by the National Health Commission of China. All the patients were laboratory‐confirmed positive for SARS‐CoV‐2 by use of quantitative RT‐PCR (qRT‐PCR) of throat swab samples. Primers and probes targeting the SARS‐CoV‐2 envelope gene were used and the sequences were as follows: forward primer, 5′‐TCAGAATGCCAATCTCCCCAAC‐3′; reverse primer, 5′‐AAAGGTCCACCCGATA CATTGA‐3′; and the probe 5′‐CY5‐CTAGTTACACTAGCCATCCTTACTGC‐3′‐BHQ1. Conditions for the amplifications were 50°C for 15 min, 95°C for 3 min, followed by 45 cycles of 95°C for 15 s and 60°C for 30 s.2.22) to fraction of inspired oxygen (FiO2) ratio (PaO2/FiO2) ≤300 mmHg (1 mmHg = 0.133 kPa). Patients meeting any of the following conditions are classified as having critical disease: (1) Development of respiratory failure requiring mechanical ventilation, (2) onset of shock, and (3) presence of other organ failures necessitating ICU monitoring and treatment. The 74 patients involved in this study are severe or critically ill COVID‐19 patients, and were categorized into three groups based on treatment outcomes. Perished cases: The throat swab samples were collected three days before the patient died. SARS‐CoV‐2 virus nucleic acid is positive based on qRT‐PCR test of throat swab samples. CDVC cases: Even after 40 days of hospitalization, throat swab samples from the patients still tested positive for SARS‐CoV‐2 virus nucleic acid using qRT‐PCR. Although some clinical symptoms showed partial improvement, certain symptoms like fever and cough persisted. These cases, characterized by delayed clearance of the virus, were referred to as CDVC patients with prolonged viral presence. Additionally, the initial positive result for SARS‐CoV‐2 was detected within 3 days from the corresponding date. A total of 13 cases met these inclusion criteria and were included in our study. Recovered cases: Patients no longer had a fever for at least 3 days, respiratory symptoms were significantly improved or returned to normal, no dyspnea, percutaneous oxygen saturation (SpO2) ≥ 95% at a rest stage. CT chest imaging showed that the lung had lost the glabrous glass shape or was largely reabsorbed, no other organ displayed severe indication, and fully recovered and discharged within 7 days after the samples were collected. The SARS‐CoV‐2 nucleic acid was positive <30 days based on qRT‐PCR tests of pharyngeal swab samples, the viral nucleic acid was negative for more than 7 days when the samples were collected, and no subsequent positive tests.Based on the criteria outlined in the COVID‐19 Diagnosis and Treatment Plan (sixth edition), all patients included in this study were classified as having severe or critical disease. According to the guidelines set forth in the COVID‐19 Diagnosis and Treatment Plan (sixth edition) issued by China, patients meeting any of the following criteria are classified as having severe disease: (1) shortness of breath with a respiratory rate (RR) greater than or equal to 30 beats per minute; (2) resting oxygen saturation below 93%. Arterial partial pressure of oxygen , and chest x‐rays or CT scans for all the patients. All information was obtained and curated with a customized data collection form. Two investigators (Yi Zhang and Qinghua Qiao) independently reviewed the data collection forms to verify data accuracy.2.42). Further experimental procedures were completed under biosafety level II plus condition. Flow cytometry analysis was conducted in seven separate batches, along with biochemical and cytokine analyses. Laboratory analysis of the samples was performed on the same day or within 1 day of the flow cytometry analysis. Please note that the results of the laboratory analysis for individual patient samples may not be available on the same day as the flow cytometry analysis. The flow loop gate strategy has been shown in Figure Peripheral blood samples from patients were harvested with anticoagulants . Continuous variables were directly expressed as a range or median (interquartile range). Categorical variables were expressed as numbers/NUMBERS (%). 2.6This study was approved by the Ethics Committee of Wuhan Huoshenshan Hospital , and all patients signed informed consent.2.7Funding agencies did not participate in study design, data collection, data analysis, or manuscript writing. The corresponding authors were responsible for all aspects of the study to ensure that issues related to the accuracy or integrity of any part of the work were properly investigated and resolved. The final version was approved by all authors.33.1n = 29, 39.19%), type‐I diabetes , and cardiovascular disease . All of these patients suffered from acute respiratory distress syndrome based on available arterial blood gas data. Less common complications included respiratory failure , secondary infection , acute cardiac injury , acute kidney injury  and acute liver injury . The clinical symptoms included cough , fever , dyspnea , fatigue , myalgia , and expectoration . All of these severe COVID‐19 patients were given empirical antimicrobial treatment including moxifloxacin and/or cephalosporin, as well as antiviral therapy like oseltamivir and/or ganciclovir. Moreover, 28 (37.84%) cases received corticosteroid (methylprednisolone) during the course of hospitalization. Twenty‐four (32.43%) cases required noninvasive mechanical ventilation and 1 (1.35%) cases required invasive mechanical ventilation  cases perished, with all of these cases being male, and the median number of days from illness onset to death being 47 days. Fortunately, 52 cases (70.27%) were categorized in the recovered group based on the definition provided above. Meanwhile, there were 13 (17.57%) cases with persistently positive qRT‐PCR tests of pharyngeal swab samples for SARS‐CoV‐2 after 40 days of hospitalization, even though the clinical symptoms in some patients were alleviated. These 13 patients were categorized as the CDVC group. CDVC cases included 9 males and 4 females, with an average age of 65.92 years, 9 cases were over 60 years old, and 6 cases had coexisting disorders, and none of them had secondary infection nor acute kidney injury. All cases received antiviral therapy, 7 (53.8%) cases had also received corticosteroid therapy, and 9 (69.2%) required noninvasive mechanical ventilation. Most importantly, 6 (46.2%) cases had high concentration of serum SARS‐CoV‐2 specific IgM antibody (>50 AU/mL) and 7 cases (53.8%) were positive for SARS‐CoV‐2 specific IgG antibody (>100 AU/mL), demonstrating that significant levels of both SARS‐CoV‐2 viral‐specific antibodies were found in some CDVC cases  and IL‐8  were 357.1‐ and 14.9‐fold than the upper limit of normal range (ULN), respectively. Interestingly, serum concentrations of these cytokines were completely below ULN in the recovered group. However, in the CDVC group, although IL‐6  and IL‐8  were lower than those in the perished group, they were still higher than ULN. IL‐1β, a pro‐inflammatory factor that promotes tissue damage during viral infection,To further clarify the inflammation status of these patients, we next tested other inflammation‐associated biochemical indicators. Interestingly, most of perished cases manifested high levels of these factors, for example, enhancement of C‐reactive protein , total bilirubin , direct bilirubin , indirect bilirubin , total biliary acid , blood urea nitrogen , alkaline phosphatase , lactate dehydrogenase , α‐hydroxybutyrate dehydrogenase , procalcitonin , myoglobin , and natriuretic peptide type B  was observed in perished group, suggesting an increased level of systemic inflammation  scans of all patients on admission. The typical findings of chest CT images of severe COVID‐19 on admission showed bilateral ground‐glass opacity and subsegmental areas of consolidation, then progressed rapidly with mass shadows of high density in both lungs. Representative chest CT images of perished cases showed that the bilateral ground‐glass opacity was maintained. However, subsequent chest CT images revealed that the bilateral ground‐glass opacity had been resolved in CDVC and recovered cases Figure .10/L, range 3.1–35.8 × 109/L) in the perished group were 1.31 folds above the ULN, and these patients also had high percentage  and absolute number  of neutrophils. However, the numbers of both WBC and neutrophils in the CDVC and recovered cases returned to normal levels  is the main feature of severe COVID‐19 patients.9/L (range 0.3–0.9 × 109/L). Mild recovery of lymphocyte counts was observed in CDVC patients but the absolute number  was still below the lower limit of normal range (LLN) , and CD8+ T cells  in perished cases were significantly lower than LLN, whereas, the frequency and absolute numbers of these cells had returned to normal levels in recovered group. Enhancement of frequency and counts of CD4+ T cells , and CD8+ T cells  were also seen in CDVC cases, but the levels were still lower than recovered group  have an important inhibitory effect during viral infection.Immune cell subsets were further analyzed by flow cytometry, from which we observed that the frequency and absolute numbers of CD4p Figure , Table 3+CD45RA+ naïve CD4+ and CD8+ T cells in circulation than perished and CDVC groups ; therefore, the recovered patients also manifested higher frequency of CD4+ and CD8+ TCM than perished as well as CDVC cases  than recovered group , a polypeptide hormone produced by thymic epithelial cells, can effectively increase T cell numbers, support T cell differentiation, maturation, and reduce cell apoptosis.Our study has some limitations. First, the data regarding the viremia profile of SARS‐CoV‐2 are not available, and further studies are needed to investigate the correlation between the viral load kinetics and the dynamics of cellular immune response. Secondly, only throat swab samples were collected from all enrolled participants, and there was a lack of lower respiratory tract samples for immunological analysis. We lack information on the current form of the pandemics, such as changes in treatment or the emergence of new varieties. A recent report revealed that 22 of the 133 patients admitted with COVID‐19 had an initial or follow‐up positive sputum or fecal sample after the conversion of their pharyngeal samples from positive to negative.In conclusion, the CDVC group represents a distinct subset of patients with specific characteristics. These findings emphasize the importance for physicians to thoroughly investigate and manage these individuals. It is crucial to consider that prolonged positivity for the virus may have implications for decisions regarding isolation programs and appropriate patient care.Yuzhang Wu, Yi Zhang, and Qinghua Qiao designed the experiments. Qinghua Qiao, Xuemin Sun, Wenjiong Shi, Hao Li, and Zhenhua Zhang contributed to patient recruitment, experiments, and data collection. Jinsong Wang, Debao Li, and Bo Tang analyzed the data. Yi Zhang and Qinghua Qiao interpreted the results and wrote the article.The authors declare no conflict of interest.This study was approved by the National Health Commission of China and the Ethics Commission of Huoshenshan Hospital.


Acute lower respiratory tract infections (ALRIs) are one one of the leading causes of morbidity and mortality among people of all ages worldwide, particularly in low‐ and middle‐income countries (LMICs). The purpose of this study was to determine epidemiological characteristics of respiratory viruses in acute respiratory infection (ARI) patients during the COVID‐19 pandemic in Yaoundé, Cameroon.Patients were monitored for respiratory symptoms as part of the surveillance of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) and other respiratory viral infections. Patients of all ages with respiratory symptoms less than 5 days were considered. Sociodemographic and clinical data as well as nasopharyngeal samples was collected from patients. Nasopharyngeal samples were tested for SARS‐CoV‐2, influenza, and respiratory syncytial virus (RSV) using real‐time reverse‐transcription polymerase chain reaction methods. Virus distribution and demographic data were analyzed with R version 2.15.1.From July 2020 to October 2021, 1120 patients were included. The overall viral detection rate was 32.5%, including 9.5% for RSV, 12.6% for influenza virus and 12.8% for SARS‐CoV‐2. Co‐infections were detected in 6.9% of positive cases. While RSV and influenza virus showed seasonal trends, SARS‐CoV‐2 was detected throughout the study period.We found that during COVID‐19 pandemic, respiratory viruses play an important role in etiology of influenza‐like illness in Cameroon, and this observation was true for patients of all ages. Since then, surveillance of influenza and other respiratory pathogens including RSV has been performed in routine. This has led to better understanding of epidemiology of major respiratory pathogens in the country through numerous studies conducted and published.22.1Administratively, Cameroon is organized into 10 regions. Among these regions is the Central region with Yaoundé as its capital. Despite climatic changes observed in recent years, this region is often characterized by an equatorial Guinean type climate with four seasons: two rainy seasons and two dry seasons.We conducted a descriptive study from March 2020 to October 2021. This study was carried out on one hand in 05 influenza sentinel sites in Yaoundé city established for influenza surveillance since 2007 and in the other hand, at CPC, which was established for surveillance of SARS‐CoV‐2 in 2020. These sentinel sites composed of public (three) and private (two) primary care centers as well as CPC are all located in Yaoundé city, Centre region of Cameroon. Patient's recruitment happened differently between sites and CPC. As far as sentinel sites are concerned, qualified medical personnel identified all influenza‐like illness (ILI) and severe acute respiratory infection (SARI) cases presenting daily in hospitals. ILI or SARI case was determined using a WHO case definition. An ILI case is an ARI with the following symptoms: fever measured ≥38°C, cough, with onset within the past 10 days, whereas a SARI case is an ARI with the following symptoms: fever or fever measured ≥38°C, cough, with onset within the past 10 days and requiring hospitalization.2.2This study was reviewed and approved by the Centre Regional Human Health Research Ethics Committee and Cameroon Ministry of Public Health.2.3Sociodemographic and clinical information was obtained from participants using a standardized questionnaire.For samples originating from influenza sentinel sites, nasopharyngeal and/or oropharyngeal swab specimens were collected from all included patients and transported to CPC in cooler containing ice packs. At CPC, total RNAs were extracted using the QiAMp Viral RNA Extraction Kit as previously described.2.4t‐test, Mann–Whitney U‐test, and Kruskal–Wallis rank test were used for description of continuous variables, and Fisher's exact test was used for description of categorical variables. We analyzed clinical characteristics and status of participants with respect to the presence or absence of respiratory infection using a stepwise logistic regression model. p‐values <0.05 were considered statistically significant for all parameters. Uncertainty was expressed as 95% confidence intervals (CIs). Statistical analysis was carried out with R program version 2•15•1.Information from the questionnaire and laboratory results were saved in a Microsoft® Excel database . We analyzed demographic and clinical characteristics of study subjects, as well as frequency and seasonal patterns of the three respiratory viruses. Student's 33.1During the period from July 2020 to October 2021, 1120 patients with ILI or SARI were included   and 141/1120 (12.6%), respectively, followed by RSV with a frequency of 106/1120 (9.5%). Of the 141 influenza viruses detected, 127 (90.1%) were influenza A viruses (80 H1N1 [A (H1N1) pdm09], 47 A [H3N2]), and 14 (9.9%) were influenza B viruses , followed by influenza A virus  and SARS‐CoV‐2 . On the other hand, in children aged 1–4 years, influenza A virus was the most frequently detected virus , followed by RSV , and there were no cases of SARS‐CoV‐2 detection. However, in age groups ≥5 years, SARS‐CoV‐2  was the most frequently detected virus with higher detection rates  in the age group 30–65, followed by influenza A virus  and then RSV .p = 0.017, odds ratio [OR], 12.48; 95% CI, 1.55–99, 98), whereas SARS‐CoV‐2 positive specimens were only negatively associated with myalgia . No other clinical symptoms were associated with positive patients, nor with any other specific virus detected or co‐infection.In a downward Wald binary logistic regression, RSV positive specimens were negatively associated only with rhinorrhea (Data are no. (%) of patients.3.3All three viruses circulated were observed throughout the study period. Figure 4This study is the first to describe the frequency and distribution of Respiratory Syncytial Virus and influenza virus associated with ILI patients in Cameroon during the COVID‐19 pandemic. The data presented here are unique to our knowledge. Respiratory viruses were detected in 32.5% of samples; SARS‐CoV‐2 and influenza virus were the most frequently detected viruses. Almost moderately high frequency of virus detection in patients with ILI during the pandemic in our study is consistent with results of other etiological studies conducted in similar settings,As in many other studies, SARS‐CoV‐2 was the most common viral pathogen detected in ILI patients, representing 12.8% viruses detected. This value is lower than that  reported in most other similar studies.During the first year of the pandemic, influenza surveillance system was greatly affected as the same actors were requisitioned to perform SARS‐CoV‐2 surveillance, and thus, there was a marked decrease in activities and consequently decreasing influenza positivity rates. Influenza positivity rate of 12.6% was observed during this study, which is lower in 2007; CPC was designated NIC by the Ministry of Public Health, Cameroon, and also lower than a study that aimed to describe 10 years of influenza activity in Cameroon.Identifying multiple respiratory virus co‐infections can provide insight into different clinical symptoms, long‐term health effects, and appropriate prevention methods. In case of viral respiratory tract infections leading to pneumonia, mixed infection can lead to severe illness in patients with compromised immune systems.Pandemic development is dynamic and dominated by immunological naivety world population to SARS‐CoV‐2. This study found that SARS‐CoV‐2 infections in Cameroon did not have an overall distribution similar to influenza and RSV. Although SARS‐CoV‐2 is the most frequently detected virus in this study, higher rates were observed in March 2020 and April 2021. In general, these three respiratory viruses exhibited different temporal distribution during study period. The observed distribution differed from that reported in published Cameroonian studies describing pathogens other than SARS‐CoV‐2.This study is not without limitations. First, our study included people that were tested at CPC laboratory and therefore is not representative of all people tested in Yaoundé. Secondly, we tested only three RNA respiratory viruses, no DNA respiratory viruses, and no respiratory bacteria or fungi. This would have limited full spectrum detection of present seasonal respiratory viruses and underestimated viral and/or bacterial co‐infection in patients with ILI. Third, SARS‐CoV‐2 virus testing methods have changed over time to meet pandemic needs. Differences in sensitivity and specificity between tests may exist, which may have caused some missed detections of SARS‐CoV‐2 and consequently, fewer co‐infection detections. Finally, we did not have access to all patient records and therefore relied on clinical information provided on the form, which may have prevented full exploration of disease severity.Despite these limitations, this study is first to describe etiology of RNA viral pathogens involved in upper respiratory tract infections during COVID‐19 pandemic in Cameroon. These viruses can also induce more serious diseases, especially in patients of all ages, which must be studied.This study should be applied in various settings and over many years to determine seasonal variation of ALRIs and individual viruses with the aim of providing a better understanding of differences in seasonality, epidemiology, and spectrum of disease caused by respiratory viruses in Cameroon. However, whether concurrent viral infection in patients with SARS‐CoV‐2 can potentially lead to viral interference or impact disease outcome awaits further research.Moïse Henri Moumbeket Yifomnjou: Formal analysis; methodology; writing–original draft; writing–review and editing. Gwladys Chavely Monamele: Investigation; writing–review and editing. Mohamadou Njankouo‐Ripa: Investigation; writing–review and editing. Abdou Fatawou Modiyinji: Investigation; writing–review and editing. Paul Alain Ngoupo: Investigation; writing–review and editing. Boyomo Onana: Conceptualization; methodology; supervision; writing–review and editing. Richard Njouom: Conceptualization; funding acquisition; methodology; supervision; validation; writing–review and editing.The authors declare that they have no competing interests.https://www.webofscience.com/api/gateway/wos/peer-review/10.1111/irv.13131.The peer review history for this article is available at