Article Type: Letter to the Editor

Impaired humoral immunity of COVID-19 patients with re-detectable positive RNA test after recoveryQing Lei1,2, Caizheng Yu3, Yang Li4, Hongyan Hou5, Banga Ndzouboukou Jo-Lewis1, Zhuqing Ouyang1, Yandi Zhang1, Xiaosong Lin1, Zongjie Yao1, Fu hui1, Ziyong Sun5, Feng Wang5,*, Shengce Tao4,*, Xionglin Fan1,*1Department of Pathogen Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.2Department of Medical Laboratory, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.3Department of Public Health, Tongji Hospital, Tongji Medical College, Huazhong University ofScience and Technology, Wuhan, China.4Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai 200240, China5Department of Laboratory Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.*Correspondence’ e-mail: Xiong-lin Fan(xlfan@hust.edu.cn), Sheng-ce Tao(taosc@sjtu.edu.cn), Feng Wang(fengwang@tjh.tjmu.edu.cn)To the Editor,The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has rapidly spread worldwide.1,2As of May 28, 2020, WHO estimated that there had been 169 million confirmed cases and 3.5 million patients died from SARS-CoV-2 infection.3 Due to lack of etiological treatments, about 3.5%-18% of discharged COVID-19 patients after recovery had a re-tested positive result for SARS-CoV-2 RNA, who was also named after re-detectable positive patients (RP).4-8 Moreover, RP patients may have the potential for virus transmission.9,10However, the mechanism of the occurrence of RP patients remains little known.Humoral immunity, especially neutralizing antibody plays an important role in preventing against SARS-CoV-2 infection, and even disease progression. To elucidate the role of humoral immunity in RP patients, we compared anti-SARS-CoV-2 IgM and IgG antibody responses between RP and non-RP patients in this study. 21 RP patients from Wuhan Pulmonary Hospital (Wuhan, China) between Jan 11, 2020, and April 1, 20204 and 113 non-RP patients discharged from Tongji Hospital, between February 17, 2020, and April 28, 2020 were recruited. Non-RP had negative NAT results while RP patients had at least once positive result during weekly follow-up after discharge. The median time from discharge to a positive retest of 21 RP patients was 14 (range: 4-24) days, and their serum samples were collected at the time of the positive retest. Besides, the serum samples of 113 non-RP patients were collected on the 14th day after discharge. At the same time, we collected serum samples of 601 non-COVID-19 controls including healthy donors, lung cancer patients, patients with autoimmune diseases from Ruijin Hospital (Shanghai, China), Tongren Hospital (Shanghai, China), or National Institutes for Food and Drug Control.11 All serum samples were stored under -80oC until use.First, we used a homemade proteome microarray to detect serum IgM (red) or IgG (green) antibody responses against 20 out of 28 predicted proteins of SARS-CoV-2 with Alexa Fluor 647-conjugated donkey anti-human IgM and Cy3-conjugated goat anti-human IgG (Figure S1) , as previously described. Based on the fluorescent intensity extracted from the microarray, anti-SARS-CoV-2 IgM and IgG profiles for each serum sample were established (Figure 1 ). Interestingly, there were differential IgM and IgG profiles observed between RP patients and non-RP patients. RP patients elicited lower levels of IgM and IgG responses against NSP7, ORF3a, and ORF7b, as well as anti-S1 and N specific IgM and IgG than non-RP patients (Figure 2 ). More importantly, RP patients induced higher levels of specific IgG antibodies against most non-structural and accessory proteins, such as NSP1, NSP2, NSP4, NSP5, NSP7, NSP8, NSP9, NSP10, RdRp, NSP14, NSP15, NSP16, ORF-3b, ORF6, and ORF9b, and E structural protein (Figure 2 ). Previously, the levels of IgG responses to NSP4, NSP7, NSP9, NSP10, RdRp, NSP14 and ORF3 might be related with the poor prognosis and severity of COVID-19.Further, a pseudotyped virus-based neutralization assay platform was used to detect the levels of serum nAb. Unexpectedly, RP patients elicited lower levels of nAb than non-RP. In particular, 90.5% (19/21) RP patients could not detect the nAb response (NT50<10) (Figure S2 ).In conclusion, RP patients impaired humoral immunity, which might benefit the replication of residual SARS-CoV-2 virus in vivo. Our findings have an important impact on improving clinical management. The criterion for patients’ recovery and discharge should be improved to prevent recurrence , such as serological detection and even nAb response, besides persisting and repeated NAT screening.

Peptides play a key role in controlling many physiological and neurobiological pathways. Many bioactive peptides require a C-terminal α-amide for full activity. The bifunctional enzyme catalyzing α-amidation, peptidylglycine α-amidating monooxygenase (PAM), is the sole enzyme responsible for amidated peptide biosynthesis, from Chlamydomonas reinhardtii to Homo sapiens. Many neuronal and endocrine functions are dependent upon amidated peptides; additional amidated peptides are growth promoters in tumors. The amidation reaction occurs in two steps, glycine α-hydroxylation followed by dealkylation to generate the α-amide product. Currently, most potentially useful inhibitors target the first reaction, which is rate-limiting. PAM is a membrane-bound enzyme that visits the cell surface during peptide secretion. PAM is then used again in the biosynthetic pathway, meaning that cell-impermeable inhibitors or inactivators could have therapeutic value for the treatment of cancer or psychiatric abnormalities. To date, inhibitor design has not fully exploited the structures and mechanistic details of PAM.

Allergen exposure chambers (AEC) provide controlled allergen exposure to allergic subjects for the clinical study of asthma and allergy. They should ideally mimic natural allergen exposure, and provide better control of allergen exposure than possible in field studies. For AEC exposure to cat allergens, typically, either liquid allergen extract is nebulized, or natural cat hair and dander are aerosolized by shaking cat bedding. While bedding shaking is more naturalistic than liquid extract exposure, it results in high variability of allergen levels. We have developed an automated method of natural dander dispersal that uses robotic vacuum cleaners with filters removed and modified for variable suction. The system was validated in two rooms (14.4 m3 and 36.7 m3) where two cats reside. The vacuums aerosolize aspirated dander that has naturally accumulated on the floor. Dispersion was characterized by measured airborne allergen (Fel d 1) and particle sizes and concentrations in time and space during 1 and 2-hour tests. At optimized parameters, Fel d 1 was found to be stable in time (2 hours), and homogenous throughout the rooms. Average Fel d 1 was 55 (±9 SD) ng/m3 in the smaller room and 79 (±30 SD) ng/m3 in the larger room, which are comparable to exposure in homes with cats. This novel method of dander dispersal provides controlled, safe exposure to cat allergens in a clinical setting, while maintaining the naturalistic advantages of a field exposure.

Background: Identification of gene polymorphisms and SNP analysis of individual patients have become crucial to select the best drug response and ensure optimal care, especially for genes directly related to asthma, such as CYSLTR1 and GSDML gene polymorphisms, an efficient and accurate detection method is helpful for the development of precision therapy. The novel detection kit we used has a reading accuracy of 99.999% for each base and the cost of detection is only 80 minutes. Objectives: In this study, a novel kit produced by Wuhan healthcare Biotechnology Co., LTD was used to detect CYSLTR1 and GSDML gene polymorphisms in asthma patients and the reliability of the novel kit was verified. Methods: The efficiency of the new kit was validated compared to gold standard, gene sequencing methods, and the close association of CYSLTR1 and GSDML SNPs with allergic asthma was also tested. As a result, the most effective drugs were obtained for patients with different gene locus expressions. Results: Identifying 367 publication participants for analysis by two different methods, we validated the reliability of the reagents with “gold standard” assays in full agreement with the DNA direct sequencing method. Our study focused on CYSLTR1 and GSDML gene polymorphisms to select the most effective drugs for asthma treatment, with leukotriene modulators being more effective in patients with the AA genotype of CYSLTR1 rs320995 and β2 agonists being more effective in patients with the TC genotype of CYSLTR1 rs320995.

Rationale, aims and objectives Financial conflicts of interest (FCOI) between pharmaceutical companies (Pharma) and healthcare domains may unduly influence physician-led clinical practice and patient-centered care. However, the extent of awareness and perceptions of FCOI among Japanese cancer patients remains unclear. This study aimed to assess these factors and their impacts on physician trustworthiness among Japanese cancer patients. Methods A cross-sectional study using self-administered surveys was conducted on a Japanese cancer patient advocacy group with 800 registered members from January to February 2019. Main outcome measures included awareness and perceptions of physician-Pharma interactions, their impact on physician trustworthiness, and attitudes towards FCOI among professions. We also performed thematic analyses on additional comments responders provided in the surveys. Results Among the 524 invited members, 96 (18.3%) completed the questionnaire. Of these, 69 (77.5%) were cancer patients. The proportion of participants aware of such interactions ranged from 2.1% to 65.3%, depending on the interaction type. Participants were generally neutral on how the interactions would affect physician trustworthiness. A large proportion of participants agreed that these interactions were unethical, could influence physicians’ prescribing behavior leading to unnecessary prescriptions, and negatively affect physician trustworthiness. Qualitative responses (n=56) indicated that patients expected physicians to use sound ethical judgment and avoid accepting incentives. Participants were also concerned about their treatment and the undue influence of FCOI on physicians. Conclusion Most participants were aware of at least one FCOI between Pharma and physicians and perceived them negatively. Further efforts to regulate FCOI appear necessary to protect patient-centered care.

i) Rationale, aims and objectives Evidence-based practice (EBP) is a well-known core healthcare competence, but the clinical implementation is complex. To be able to facilitate EBP implementation, valid measurement of the “EBP-status quo” is essential. Therefore, we aimed to identify valid tools for EBP status assessment among health practitioners in Germany. ii) Methods The databases MEDLINE (via Pubmed), Cochrane and PEDro were systematically searched until July 2019, to update a previous review from 2013. Methodological quality and evidence level was scored by two independent raters via: i) COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) checklist, ii) updated criteria for good measurement properties, and iii) modified GRADE criteria. iii) Results Overall, 3.467 studies were identified with 20 different EBP-questionnaires included in 38 studies on EBP tool development and/or validation. Most questionnaires demonstrated moderate or low-quality evidence for the psychometric properties tested. While validity properties (content, structural and cross-cultural) were assessed more frequently, reliability was tested in 20% of the questionnaires and responsiveness in one tool. iv) Conclusion Although a lack of high-quality psychometric properties of EBP-tools became apparent, the “Evidence-Based Practice Inventory” (EBPI) appears to be the best validated and reliable tool for a survey among German health practitioners.

Background and purpose: Ferroptosis and parthanatos are two types of programmed cell death associated with cerebral ischemia. There is a sizeable interest in seeking chemical component for the regulation of ferroptosis and parthanatos. Anhydrosafflor yellow B (AHSYB) and hydroxysafflor yellow A (HSYA) mitigated cell death caused by oxidative stress due to antioxidant capacity, yet the mechanism is still uncertain. Thus, we investigated whether AHSYB and HSYA prevent death through these two pathways with the aim to elucidate their potential protective mechanisms of cerebral ischemia. Experimental approach: Oxidative stress model was established by treating PC12 cells with oxygen glucose deprivation and reperfusion (OGD/R). Cellular functions and signaling pathways were analyzed in PC12 cells using cell counting kit-8 (CCK-8), ELISA, transmission electron microscopy (TEM), immunofluorescence, and western blot. Key results: HSYA and AHSYB protected cells from oxidative stress. The phenomenon is associated with ferroptosis and parthanatos. HSYA and AHSYB upregulated cystine/glutamate antiporter system xc- (system xc-) and glutathione peroxidase 4 (GPX4), returned the GSH/GSSG ratio and reactive oxygen species (ROS) to homeostasis, alleviated lipid peroxidation. By reason of reducing ROS, HSYA and AHSYB restrained poly(ADP-ribose) polymerase-1 (PARP-1) overactivation, reduced the production of excess poly(ADP ribose) (PAR) polymer and apoptosis inducing factor (AIF) nuclear translocation. Conclusions and implications: The results suggested that HSYA and AHSYB limited ferroptosis and parthanatos to alleviate oxidative stress in PC12 cells. These findings may have implications for improved understanding of how drugs reduce oxidative stress and develop new strategies for treating degenerative diseases such as cerebral ischemia.

Background: The histopathology of pediatric chronic rhinosinusitis with nasal polyps (CRSwNP) and without nasal polyps (CRSsNP) is rarely reported due to their low prevalence or the unavailability of tissue samples. Hence, we aimed to characterize and compare the histologic features and protein expression of Th1/Th2/Th17-related cytokines in pediatric CRSsNP and CRSwNP. Methods: The histologic characteristics of 15 children with CRSsNP, 52 children with CRSwNP, and 12 control participants were analyzed using hematoxylin and eosin staining. The expression of Th1/Th2/Th17-related cytokines were examined using immunohistochemistry and the enzyme-linked immunosorbent assay. Results: Pediatric subjects with CRSwNP had more intact epithelium and less submucosal mucous glands compared to those with CRSsNP. Tissue eosinophils were more prevalent in the young CRSwNP group compared to the old CRSwNP or the CRSsNP groups. The protein concentrations of Th2 cytokines were significantly higher in the CRSwNP group than the CRSsNP group or the control group. Moreover, the protein concentrations of Th17 cytokines were significantly higher in the young CRSwNP group than the old CRSwNP group or the CRSsNP and control groups. The protein concentrations of Th1 and Th17 cytokines were also significantly higher in the CRSsNP group than the control group. Compared with non-eosinophilic CRSwNP, eosinophilic CRSwNP presented with elevated protein concentrations of Th1 and Th17 cytokines. Conclusion: For the first time, we showed that pediatric CRSwNP presents as eosinophilic with Th2/Th17 inflammation, whereas CRSsNP presents as Th1/Th17 inflammation. Our study may provide a theoretical basis for the precise treatment of pediatric CRS in the future.

Chronic spontaneous urticaria (CSU) is defined as the recurrent episodes of spontaneous wheals and/or angioedema for more than 6 weeks, and at least twice a week. The core link in the pathogenesis of CSU is the activation of mast cells and other immune cells caused by various reasons. These activated immune cells release a series of inflammatory active mediators such as histamine, arachidonic acid metabolites, chemokines, etc., resulting in clinical features such as wheals and/or edema. However, the specific mechanisms leading to the activation of various immune cells have not been fully elucidated. Previous evidence has shown that about 50% of CSU patients have potential autoimmune reactions. Here we review the functional implication of immune cells in CSU focusing on the crosstalk between them and discuss whether their crosstalk promotes the occurrence and development of the disease.

Background and Purpose: Post-traumatic epilepsy is the confirmed progression of unprovoked seizures after head trauma in the general population. This study was designed to control the devastating chronic consequences of post-traumatic epileptogenesis. Experimental Approach: Cerebral trauma was experimentally induced in Wistar rats to challenge the seizure susceptibility for futuristic PTE. The efficiency of Flufenamic acid, and fasudil hydrochloride, were assessed and compared with valproic acid for abnormal neurobehavioural symptoms, biomolecular levels, and apoptotic changes through histopathology. Key Results: Initial brain insult was successfully exacerbated the normal pathophysiology and neurobehavior changes in an experimental model. The lowered seizure threshold confirmed the epileptogenesis progression when a jab of Pentylenetetrazol(35mg/kg), gave the first successive seizure. Disrupted BBB permeability, neuronal infarction, and brain edema were found restored to normal after the treatment when compared to disease controls. The altered levels of neurotransmitters, mitochondrial complexes, cytoplasmic biomolecules/cytokines, different genes, and channel proteins were also restored to normal after 14-days treatments. No seizures have been observed with the test dose of Pentylenetetrazol. We analyzed the histopathological changes of hippocampus/cortex regions and found a significant decrease in the count of non-evident nucleoli, nuclear pyknosis, vascular congestion, and perineural vacuolization in treatment groups compared to injury controls. Conclusion and Implications: All the drugs greatly minimized the epileptogenesis cascade of futuristic PTE, confirms their neuroprotective capabilities. Flufenamic acid was found more effective with many potential abilities to minimize epileptogenesis compared to fasudil hydrochloride but its combination with valproic acid has much neuroprotection and efficacy.

Background: The COVID-19 pandemic led to an increase in research activity worldwide. The Vall d’Hebron University Hospital Research Ethics Committee (VH-REC) adapted its procedures to give out the opinion rapidly. We aimed to describe the characteristics of the VH-REC activity and studies evaluated during the first outbreak. Methods: Clinical trials (CT), post-authorization studies (PAS) and research projects (RP) on COVID-19 were included and followed up. Variables were described through usual descriptive methods. Results: 157 studies were evaluated: 10 CT, 16 PAS and 131 RP, in 25 bi-weekly online meetings. Non-commercial, unicentric and national studies predominated (95%, 54% and 88%, respectively). The main objective of CT and PAS studies was to test efficacy and safety, and for RP to describe patients’ outcomes. Some studies focused on specific interest groups, such as healthcare professionals or immunosuppressed patients (10% both). The median time of protocols’ evaluation was 3 days. 58.6% (92) required further clarifications, mainly due to aspects of data protection, informed consent, and biological samples. The final opinion was favourable in 93% (146). Regarding follow-up, 123 studies had been initiated and 64 also finalized. Results have been published in 59% (51) of studies. Conclusions and implications: COVID-19 pandemic has led to greater academic and local research, especially through research projects. Electronic sources were implemented for evaluation shortening and ease follow-up. These measures should remain to streamline VH-REC processes, and trends to publish results favoured. This study could allow comparisons with other activity periods (e.g. pre or post-pandemic), or with other REC’

The foremost aim of this paper is to reveal the extent of the controllability of the semilinear evolution integrodifferential impulse system with delayed impulses and non local conditions. This paper begins with the grit of the control formula for the same impulse system in Banach space. Moreover, As a result, is extended to controllability. The sufficient conditions are introduced by utilizing the Hausdorff measure of noncompactness, Sadovskii fixed point theorem, and operator semigroups in appropriate dropping compactness of the operator. Sequentially, an example is provided to show our results.

Background: Telemedicine uses technology to deliver medical care to patients at different locations. Teleregulation (TR), a type of telemedicine, occurs when a specialist evaluates data from a general medical doctor. The aim of the current study is to analyze the use of asynchronous TR for patient referral, from primary care to a neurology specialist, in the city of Curitiba in Southern Brazil. Method: A retrospective analysis of all patients referred from primary care to neurology between September 2019 and February 2020. After a request is made by a general medical doctor for a specialist’s opinion, five neurologists with complete access to patients’ records are tasked with the decision-making. The main variables analyzed were: clinical reasons for enrollment to the TR, the TR decision, final diagnosis, indication for diagnostic procedures, and subsequent follow-up. Results: Between September 2019 and February 2020, 1035 TR sessions were performed. Headache (30.43%), epilepsy (19.03%), and dementia (15.85%) accounted for almost two-thirds of the primary care requests. One-third of the cases (33.62%) required a complementary diagnostic procedure. More than 70% of the cases did not require face-to-face assessment by a neurologist. Discussion and conclusion: Neurology is a specialty in high demand and some pathology tests require rapid decision-making. In this study, asynchronous teleneurology successfully reduced the need for in-visit consultation in 70% of cases. Further studies should identify the best opportunities for teleneurology in the city of Curitiba to facilitate better integrated care between primary care providers and neurologists.

Aim: In this study, reported suicide cases between 2013 to 2017 in Turkey was investigated to the interest of marital status, gender, educational attainment, and causes. Material and Methods: Data was performed by using the Turkish Statistical Institute National Databases. The study variables analyzed were years, sex, cause of suicide, educational attainment, and marital status. Chi-square analysis, correlation, and regression analysis were used to compare outcomes of suicides among study variables. Results: The most common causes of suicide among people who live in Turkey were, unknown reasons, illness, other reasons and, economic problems. When we examine total suicides number by educational level respectively; Primary school, primary education, and then high school and equivalent attainment were more; on the other hand by legal marital status married and never married persons were found mostly. The level of education (P=0,000) and marital status (P=0,001) of the people were influential on suicide. Discussion: Without forgetting that the causes of suicides’ are been multifactorial; marital status and educational attainment may affect suicides.

Trefoil factor 3 (TFF3) is the last small molecule peptide found in the trefoil factor family. It is mainly secreted by intestinal goblet cells and exerts mucosal repair effect in gastrointestinal tract. Emerging evidence indicated that TFF3 is not only expressed in intestinal tract, but also in female reproductive, urinary, respiratory, immune, nervous and endocrine system. Although TFF3 is widely distributed in human healthy samples, its specific physiological significance is unclear. This review provides an up-to-date overview of the biological functions of TFF3 and summarizes its expression sites, binding proteins, signaling pathways and clinical applications. We found that TFF3 mediated signaling pathways mainly include MAPK, NF-κB, PI3K, STAT3 and HIF-1. More importantly, mucosal protection would no longer be the only application of TFF3. As TFF3 gradually exhibits the potential for immune regulation, neuroprotection, and regulation of glycolipid metabolism, it may get many interesting clinical applications in the future.

Background and Purpose. Heart failure (HF) therapeutic toolkit would strongly benefit from the availability of ino-lusitropic agents with a favorable pharmacodynamics and safety profile. Istaroxime is an agent, combining Na+/K+ pump inhibition and SERCA2a stimulation, shown by phase 2 trials to be promising in the acute setting. As PST3093 is an istaroxime metabolite reaching plasma concentration and duration greater than those of istaroxime, its evaluation is indispensable to establish its contribution to the istaroxime clinical efficacy. Experimental Approach. We studied PST3093 for its effects on SERCA2a and Na+/K+ ATPase activities, Ca2+ dynamics in intact myocytes and hemodynamic effects in an in-vivo rat model of diabetic (streptozotocin (STZ)-induced) cardiomyopathy. Key Results. The results converge to identify PST3093 as a “selective” (i.e. devoid of Na+/K+ pump inhibition) SERCA2a activator, thus differentiating it from its parent compound istaroxime. In in-vivo echocardiographic assessment, PST3093 improved overall cardiac performance (stroke volume and cardiac output) without decreasing heart rate, thus reversing many of STZ-induced abnormalities. Modulation of both systolic and diastolic indexes contributed to the improvement. For i.v. administration, PST3093 toxicity was considerably lower than that of istaroxime and its evaluation against a panel of 50 targets commonly involved in cardiac and extracardiac side-effects, failed to reveal significant interactions. Conclusion and Implication. PST3093 is a “selective” SERCA2a activator, the prototype of a novel pharmacodynamic category with a potential in the ino-lusitropic approach to HF therapy that may contribute to the clinical efficacy of istaroxime infusion.

Objective: There is an ongoing debate about the best and comfortable way to administer surfactant. We hypothesized that uninterrupted respiratory support and continuous PEEP implementation while instilling surfactant via endotracheal tube (ETT) with side port will result in higher regional cerebral tissue oxygenation (rcSO2) and the alterations in cerebral hemodynamics will be minimal. Methods: Preterm infants who required intubation in the delivery room and/or in the first 24 hours of life with gestational age <32 were enrolled. Patients were intubated either via conventional ETT or ETT with side port (Vygon®) with appropriate sizes. Following NICU admission a NIRS probe placed on the forehead and each infant were monitored with NIRS.In conventional ETT group, patients separated from the ventilator while surfactant was instilled. In ETT with side port group respiratory support was not interrupted during instillation. Heart rate, oxygen saturations, rcSO2, cerebral fractional tissue oxygen extraction (cFTOE) and blood pressures were recorded. Results: A total of 46 infants analyzed. Surfactant was instilled with conventional ETT in 23 and ETT with side port in 23 infants. Birthweights (1037±238 vs 1152±277g) and gestational ages (28±2.3 vs 29±1.6weeks) did not differ between groups. During instillation of surfactant, rcSO2 levels [61.5 (49-90) vs 70 (48-85)] and cFTOE levels 0.28 (0.10-0.44) vs 0.23 (0.03-0.44)] were similar (p=0.58 and 0.82 respectively). Conclusion: Interruption of respiratory support during surfactant instillation did not significantly alter the cerebral tissue oxygenation. These results did not support our hypothesis possibly due to small sample size and should be confirmed with further studies.

Complete surgical removal of a large retrovesical pelvic cyst with an unusual extension could be technically demanding with the high risk of injury to the adjacent vital structures. Opting a conservative approach may reduce morbidity, as the reported incidence of malignancy is very low. This operative technique demonstrates that no evident arising point could be identified on diagnostic laparoscopy, hence the cyst was drained vaginally with partial excision and marsupialization. Histological analysis confirmed a benign epidermoid cyst and there is no recurrence to date.

Background and Purpose: Pro-androgenic substances are often used to treat muscle- or bone-related disorders. Their interactions with the classical androgen receptor (AR), however, trigger a series of undesirable effects. It would be of great benefit if the positive androgenic effects could be obtained by circumventing the classical AR. Experimental Approach: ZIP9 is a recently identified membrane-bound androgen receptor of physiological significance. Using in silico methods we identified and verified the extracellular localization of its androgen binding site and designed small peptides that fit in it without interacting with the AR. Six tetrapeptides with the best docking properties were synthesized and further investigated. Key Results: All tetrapeptides displaced T-BSA-FITC, a membrane-impermeable testosterone analog, from the surface of mouse myogenic L6 cells that express ZIP9 but not the AR. Silencing the expression of ZIP9 with siRNA prevented this labeling. All tetrapeptides act pro-androgenic: in L6 cells they stimulated the expression of myogenin, triggered activation of focal adhesion kinase, and prompted the fusion of L6 myocytes to syncytial myotubes. In human osteoblastic SAOS-2 cells that express AR and ZIP9 they reduced the expression of alkaline phosphatase and stimulated mineralization. These latter effects were prevented by silencing ZIP9 expression, indicating that the osteoblast/osteocyte conversion is exclusively mediated through ZIP9. Conclusions and Implications: Our results demonstrate that the synthetic tetrapeptides, by acting as ZIP9-specific androgens, have the potential to replace testosterone or testosterone analogs in the treatment of bone- or muscle-related disorders by circumventing the undesirable effects mediated through the classical AR.

Background and Purpose Idiopathic pulmonary fibrosis is a progressive fatal disease characterized by interstitial remodeling, with high lethality and a lack of effective medical therapies. Tetrandrine has been proposed to present anti-fibrotic effects, but the efficacy and mechanisms of tetrandrine against lung fibrosis has not been systematically evaluated. We sought to study the potential therapeutic effects and mechanisms of tetrandrine in lung fibrosis. Experimental Approach The anti-fibrotic effects of tetrandrine were evaluated in bleomycin-induced mouse models and TGF-β1-stimulated murine lung fibroblasts. We performed Chromatin Immunoprecipitation (ChIP), Immunoprecipitation (IP) and mRFP-GFP-MAP1LC3B adenovirus construct to investigate the novel mechanisms of tetrandrine-induced autophagy. Key Results Tetrandrine decreased TGF-β1-induced expression of α-smooth muscle actin, fibronectin, vimentin and type 1 collagen and proliferation in fibroblasts. Tetrandrine restored TGF-β1-induced impaired autophagy, accompanied by the up-regulation and enhanced interaction of SQSTM1 and MAP1LC3-Ⅱ. ChIP studies revealed that NRF2 bound to SQSTM1 promoter in tetrandrine-induced autophagy. Furthermore, TGF-β1-induced phosphorylated mTOR was inhibited by tetrandrine, with reduced activation levels of Rheb. In vivo tetrandrine suppressed the bleomycin-induced expression of fibrotic markers and improved pulmonary function. Conclusion and Implications Our data suggest that tetrandrine might be recognized as a novel autophagy inducer, thus attenuating lung fibrosis. Tetrandrine should be investigated as a novel therapeutic strategy for IPF.

ABSTRACT Purpose Kidney involvement is frequent among patients with coronavirus disease 2019 (Covid-19). However, kidney involvement is varied and mild kidney injury can easily go unnoticed. We aimed to investigate the urinalysis data of Covid-19 patients on admission and to explore the value of urinalysis in the prediction of AKI and in-hospital mortality in patients with Covid-19. Methods The demographic, clinical and laboratory data of patients with confirmed Covid-19 were collected from the electronic health records of the hospital. The outcomes were development of AKI and in-hospital mortality. Results 244 patients were included in the analysis. Mean age was 59.6 ± 13.7 and 65.2% of patients were male. Median SCr on admission was 0.86 (0.72-1.05) mg/dL. Glucosuria, proteinuria and hematuria were found in 36.1%, 22.9% and 22.1% of patients, respectively. AKI was detected in 63 patients (25.8%) on any time of hospitalization. According to multivariate binary logistic regression analayses; AKI development was associated with higher WBC and decreased eGFR as well as with proteinuria on admission. During median 8 (IQR, 5-12) days of follow-up, 33 patients (13.5%) died. Older age, higher CRP levels and proteinuria on admission were also independent predictors of in-hospital mortaliy. Conclusion Proteinuria on admission is associated with development of AKI and in-hospital mortality in patients with Covid-19. Urinalysis can be useful for the evaluation of COVID-19 progression and early diagnosis of kidney damage before SCr rise.

Background: Dilated cardiomyopathy (DCM) is a condition involving dilation of cardiac chambers, which results in contraction impairment. Besides invasive and non-invasive diagnostic procedures, cardiac biomarkers are of great importance in both diagnosis and prognosis of the disease. These biomarkers are categorized into three groups based on their site; cardiomyocyte biomarkers, microenvironmental biomarkers and macroenvironmental biomarkers. Aims: In this review, an overview of characteristics, epidemiology, etiology and clinical manifestations of DCM is provided. In addition, the most important biomarkers, of all three categories, and their diagnostic and prognostic values are discussed. Conclusion: Considering the association of DCM with conditions such as infections and autoimmunity, which are prevalent among the population, introducing efficient diagnostic tools if of high value for the early detection of DCM to prevent its severe complications. The three discussed classes of biomarkers are potential candidates for the detection of DCM. However, further studies are necessary in this regard.

Purpose: Pulmonary hypoplasia remains a major cause of high mortality in congenital diaphragmatic hernia (CDH). Notch1 signaling pathway plays a critical role in alveolar epithelial cells(AECs) differentiation which dominated in the pulmonary hypoplasia of CDH. However, the effect of Notch1 pathway in CDH remains unclear. We design this study to investigate the hypothesis that the aberrant Notch1 pathway leads to the pulmonary dysplasia in nitrofen-induced CDH rat model. Methods: Pregnant rats were designed as follows: Control, CDH, CDH+CUR. Morphological, Immunostaining and qRT-PCR study were performed to determine the expression levels of T1α and TTF1 in AECs and the expression levels of gene including NICD1 and Hes1 associated with Notch1 signaling pathway. Results: Immunostaining of AEC-I marker T1α and AEC-II marker TTF1 showed that the positive area ratio of T1α in CDH group was less in CUR and Control group, TTF1 was larger in CUR and Control than CDH group (p< 0.05). NICD1 and Hes1 in CDH group were lager than CUR and Control group (p< 0.05). qRT-PCR showed that the expression levels of T1α in CDH group was lower than that in CUR and Control group (p< 0.05), and TTF1, NICD1 and Hes1 in CDH group were higher than CUR and Control group (P < 0.05). Conclusion: Aberrant Notch1 signaling pathway and AECs differentiation disorders are involved in the pathogenesis of pulmonary hypoplasia in nitrofen-induced CDH rat model. Our data suggest that overexpressed Notch1 signaling pathway may cause pulmonary hypoplasia in CDH by inhibiting the differentiation of AECs.