Real-time Reverse Transcription-polymerase Research Articles

RETRACTED: Two new coordination polymers: Magnetic properties and treatment activity on non-small cell lung cancer by reversing the resistance of the cancer cells

In the current study, by employing 5-((4-carboxypyridin-2-yl)amino)isophthalic acid (H 3 L), a semirigid carboxylate-pyridine ligand with V-shape, two new transition metal coordination polymers with the chemical formulae of [Cu 1.5 (L)(H 2 O)]·2H 2 O ( 1 ) and [Co 4 (L) 2 (CH 3 CN)(OH) 2 (H 2 O) 4 ]·3H 2 O ( 2 ) have been successfully prepared through the reaction between the H 3 L ligand and corresponding metal salts under the solvothermal reaction conditions. Magnetic investigations have suggested that there is antiferromagnetic coupling between neighboring metal ions in the two compounds. The application values of compounds 1 and 2 on the non-small cell lung cancer (NSCLC) was evaluated and the related mechanism was discussed at the same time. First of all, the Cell Counting Kit-8 (CCK-8) detection kit was performed to measure the viability of the non-small cell lung cancer cells after compound treatment. In addition to this, the real time reverse transcription polymerase chain reaction (real time RT-PCR) was used to detect the relative expression levels of the ezh2 gene in NSCLC cells after indicated treatment.

Silencing Neuropilin 1 gene reverses TGF-β1-induced epithelial mesenchymal transition in HGC-27 gastric cancer cell line*

Abstract Objective The aim of this study was to determine Neuropilin 1 (NRP1) contribution to transforming growth factor β1 (TGF-β1)-induced epithelial mesenchymal transition (EMT) of HGC-27 gastric cancer cells and study its mechanism. Methods In this study, TGF-β1 was used to induce EMT in HGC-27 cells. Further, these cells were stably transfected with siRNA targeting NRP1. Wound healing and transwell assays were used to measure cell migration and invasion, respectively. NRP1 and EMT markers were measured using quantitative real time reverse transcription polymerase chain reaction and western blotting. Results Exposure of TGF-β1 conferred a fibroblastic-like shape to cancer cells and significantly increased the expression of NRP1 in HGC-27 cells. TGF-β1 subsequently promoted migration and invasion of HGC-27 cells. Furthermore, silencing NRP1 inhibited the invasion and migration of TGF-β1-induced cells undergoing EMT. Conclusion Silencing NRP1 can inhibit cell migration, invasion, and metastasis and reverse the TGF-β1-induced EMT process of gastric cancer.

English

Zika, yellow fever and dengue viruses are three important flaviviruses that are epidemic and endemic in Central Africa, Middle and South America, Southeast Asia and Pacific Islands. Since these three viral diseases present with similar symptoms and there is a possibility of multiple flaviviruses co-infection, it is important to develop an effective and quick diagnostic tool. Here, we designed and validated a one-step triplex real time reverse transcription polymerase chain reaction (RT-PCR) using TaqMan probes targeting NS5 region for detecting zika virus (ZIKV), dengue virus-4 (DENV-4) and yellow fever virus (YFV). The assay was highly specific for the target viral RNA corresponding to ZIKV, DENV-4 and YFV strains. The sensitivity of the assay was compared to one-step, RT-PCR, it showed a 100-fold higher sensitivity than RT-PCR with ZIKV and DENV and 10-fold higher sensitivity with YFV. In conclusion, the one-step, triplex, TaqMan, real time RT-PCR assay identified and distinguished ZIKV, DENV-4 and YFV by targeting single gene. Key words: Zika, yellow fever, dengue, multiplex real time reverse transcription-polymerase chain reaction (RT-PCR).

Closed circuit system in laparoscopy: SARS-CoV-2 pandemic

Background: There is a big concern regarding transmission of Severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) during laparoscopy and using energy devices through the carbon dioxide (CO2) pneumoperitoneum. So, we decided to use a novel indigenous innovative closed-circuit system (CCS) to avoid this contamination in operation theatre.Methods: We set out to do a prospective study in the safety of this innovative closed-circuit system in elective surgeries over 3 consecutive months of May to July 2020, during the peak of SARS-CoV-2 pandemic in a tertiary teaching hospital. We recruited 52 patients undergoing elective laparoscopic surgery in a single laparoscopy unit. The Coronavirus disease 2019 (COVID-19) status was unknown, so we presumed all patients to be potential carriers of SARS-CoV-2. However, we followed the Indian council for medical research (ICMR) and Government of Karnataka (GOK) guidelines of clinical screening before surgery. We observed the postoperative outcome regarding COVID-19 symptoms in Health care workers (HCW) involved in the operation theatre. If required, we do a real time reverse transcription polymerase chain reaction (RT-PCR) to find the true positives.Results: Of the total 52 (basic and advanced) laparoscopic surgeries performed during this period, only 2 (3.84%) patients turned positive in the postoperative period. They developed COVID-19 symptoms and were tested positive by RT-PCR. None (100%) of the HCW developed any symptoms of COVID-19.Conclusion: Closed circuit system is a safe and promising technique during laparoscopy.

Coronavirus disease 2019 (COVID-19) in Italy: features on chest computed tomography using a structured report system

To assess the use of a structured report in the Chest Computed Tomography (CT) reporting of patients with suspicious viral pneumonia by COVID-19 and the evaluation of the main CT patterns. This study included 134 patients (43 women and 91 men; 68.8 years of mean age, range 29–93 years) with suspicious COVID-19 viral infection evaluated by reverse transcription real-time fluorescence polymerase chain reaction (RT-PCR) test. All patients underwent CT examinations at the time of admission. CT images were reviewed by two radiologists who identified COVID-19 CT patterns using a structured reports. Temporal difference mean value between RT-PCRs and CT scan was 0.18 days ± 2.0 days. CT findings were positive for viral pneumonia in 94.0% patients while COVID-19 was diagnosed at RT-PCR in 77.6% patients. Time mean value to complete the structured report by radiologist was 8.5 min ± 2.4 min. The disease on chest CT predominantly affected multiple lobes and the main CT feature was ground glass opacity (GGO) with or without consolidation (96.8%). GGO was predominantly bilateral (89.3%), peripheral (80.3%), multifocal/patching (70.5%). Consolidation disease was predominantly bilateral (83.9%) with prevalent peripheral (87.1%) and segmental (47.3%) distribution. Additional CT signs were the crazy-paving pattern in 75.4% of patients, the septal thickening in 37.3% of patients, the air bronchogram sign in 39.7% and the “reversed halo” sign in 23.8%. Less frequent characteristics at CT regard discrete pulmonary nodules, increased trunk diameter of the pulmonary artery, pleural effusion and pericardium effusion (7.9%, 6.3%, 14.3% and 16.7%, respectively). Barotrauma sign was absent in all the patients. High percentage (54.8%) of the patients had mediastinal lymphadenopathy. Using a Chest CT structured report, with a standardized language, we identified that the cardinal hallmarks of COVID-19 infection were bilateral, peripheral and multifocal/patching GGO and bilateral consolidation with peripheral and segmental distribution.

Thromboprophylaxis with enoxaparin is associated with a lower death rate in patients hospitalized with SARS-CoV-2 infection. A cohort study.

BackgroundSevere Acute Respiratory Syndrome CoronaVirus 2 (SARS-CoV-2) infection is associated with hypercoagulability caused by direct invasion of endothelial cells and\\or proinflammatory cytokine release. Thromboprophylaxis with enoxaparin is recommended by current guidelines, but evidence is still weak. The aim of this study was to assess the impact of thromboprophylaxis with enoxaparin on hospital mortality in patients admitted for Coronavirus disease 2019 (COVID-19). The effects of enoxaparin on intensive care admission and hospital length-of-stay were evaluated as secondary outcomes.MethodsObservational cohort study, with data collected from patients admitted to Poliambulanza Foundation with positive real time reverse transcription polymerase chain reaction (RT-PCR) for SARS-CoV-2 from 20th February to 10th May 2020. Multivariate logistic regression with overlap weight propensity score was used to model hospital mortality and intensive care admission, hospital length-of-stay was analyzed with a multivariate Poisson regression. Seven hundred and ninety nine (57%) patients who received enoxaparin at least once during the hospitalization were included in the enoxaparin cohort, 604 (43%) patients who did not were included in the control cohort.FindingsAt the adjusted analysis enoxaparin was associated with lower in-hospital mortality (Odds Ratio 0·53, 95% C.I. 0·40–0·70) compared with no enoxaparin treatment. Hospital length-of-stay was longer for patients treated with enoxaparin (Incidence Rate Ratios 1·45, 95% C.I. 1·36–1·54). Enoxaparin treatment was associated with reduced risk of intensive care admission at the adjusted analysis (Odds Ratio 0·48, 95% C.I. 0·32–0·69).InterpretationThis study shows that treatment with enoxaparin during hospital stay is associated with a lower death rate and, while results from randomized clinical trials are still pending, this study supports the use of thromboprophylaxis with enoxaparin in all patients admitted for COVID-19. Moreover, when enoxaparin is used on the wards, it reduces the risk of Intensive Care Unit admission.

The kinetics of viral load and antibodies to SARS-CoV-2

ObjectivesThe aim was to understand persistence of the virus in body fluids the and immune response of an infected host to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), an agent of coronavirus disease 2019 (COVID-19). MethodsWe determined the kinetics of viral load in several body fluids through real time reverse transcription polymerase chain reaction, serum antibodies of IgA, IgG and IgM by enzyme-linked immunosorbent assay and neutralizing antibodies by microneutralization assay in 35 COVID-19 cases from two hospitals in Guangdong, China. ResultsWe found higher viral loads and prolonged shedding of virus RNA in severe cases of COVID-19 in nasopharyngeal (1.3 × 106 vs 6.4 × 104, p < 0.05; 7∼8 weeks) and throat (6.9 × 106 vs 2.9 × 105, p < 0.05; 4∼5 weeks), but similar in sputum samples (5.5 × 106 vs 0.9 × 106, p < 0.05; 4∼5 weeks). Viraemia was rarely detected (2.8%, n = 1/35). We detected early seroconversion of IgA and IgG at the first week after illness onset (day 5, 5.7%, n = 2/35). Neutralizing antibodies were produced in the second week, and observed in all 35 included cases after the third week illness onset. The levels of neutralizing antibodies correlated with IgG (rs = 0.85, p < 0.05; kappa = 0.85) and IgA (rs = 0.64, p < 0.05; kappa = 0.61) in severe, but not mild cases (IgG, rs = 0.42, kappa = 0.33; IgA, rs = 0.32, kappa = 0.22). No correlation with IgM in either severe (rs = 0.17, kappa = 0.06) or mild cases (rs = 0.27, kappa = 0.15) was found. DiscussionWe revealed a prolonged shedding of virus RNA in the upper respiratory tract, and evaluated the consistency of production of IgG, IgA, IgM and neutralizing antibodies in COVID-19 cases.

Assessment of the frequency and biochemical parameters of conjunctivitis in COVID-19 and other viral and bacterial conditions

Abstract Objectives We compared the laboratory and clinical symptoms of coronavirus disease 2019 (COVID-19), other viral (beside COVID-19), and bacterial conjunctivitis patients with the values of controls. Methods Twenty COVID-19, 15 other viral, 15 bacterial patients, and 15 control group were included in the study. COVID-19 was diagnosed with the real time reverse transcription–polymerase chain reaction while the indirect immunofluorescent antibody test was used in the detection of other viral agents. Bacterial agents were determined with the detection of the agent. The ophthalmologic examination of all cases was carried out by direct penlight, and the anterior segment evaluation was performed. The laboratory findings of all cases included in the study were obtained from the hospital records. Results It was determined that 5% conjunctivitis and 5% diplopia developed in patients diagnosed with COVID-19, 20% conjunctivitis developed in patients due to other viral agents, and 66.6% conjunctivitis developed in patients due to bacterial agents. Conclusion The incidence of conjunctivitis in COVID-19 patients was lower than in other viral and bacterial groups. Neutrophil/lymphocyte ratio and C-reactive protein come to the forefront as precious parameters with high specificity and sensitivity that might be useful to distinguish these diseases.

Saliva is a reliable, non-invasive specimen for SARS-CoV-2 detection

BackgroundSevere Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the cause of Coronavirus Disease 2019 (COVID-19). Although Real Time Reverse Transcription Polymerase Chain Reaction (qRT-PCR) of respiratory specimens is the gold standard test for detection of SARS-CoV-2 infection, collecting nasopharyngeal swabs causes discomfort to patients and may represent considerable risk for healthcare workers. The use of saliva as a diagnostic sample has several advantages. ObjectivesThe aim of this study was to validate the use of saliva as a biological sample for diagnosis of COVID-19. MethodsThis study was conducted at Infectious Diseases Research Laboratory (LAPI), in Salvador, Brazil. Participants presenting with signs/symptoms suggesting SARS-CoV-2 infection underwent a nasopharyngeal swab (NPS) and/or oropharyngeal swab (OPS), and saliva collection. Saliva samples were diluted in PBS, followed by RNA isolation and RT-Real Time PCR for SARS-CoV-2. Results of conventional vs saliva samples testing were compared. Statistical analyses were performed using Statistical Package for the Social Sciences software (SPSS) version 18.0. ResultsOne hundred fifty-five participants were recruited and samples pairs of NPS/OPS and saliva were collected. The sensitivity and specificity of RT-PCR using saliva samples were 94.4% (95% CI 86.4–97.8) and 97.62% (95% CI 91.7–99.3), respectively. There was an overall high agreement (96.1%) between the two tests. ConclusionsUse of self-collected saliva samples is an easy, convenient, and low-cost alternative to conventional NP swab-based molecular tests. These results may allow a broader use of molecular tests for management of COVID19 pandemic, especially in resources-limited settings.

Ramping up of SARS CoV-2 testing for the diagnosis of COVID-19 to better manage the next phase of pandemic and reduce the mortality in India.

The coronavirus disease 2019 (COVID-19) pandemic is caused by the severe acute respiratory syndrome coronavirus-2, a new member of the Coronavirus family. The virus was first identified in Wuhan, China, where the epidemic originated. The viral genome was sequenced and a real time reverse transcription polymerase chain reaction assay was developed and used for the detection of virus. Different countries took different approaches for the diagnosis of COVID-19. Some countries prioritized extensive testing for COVID-19 at a very early phase of the pandemic whereas other countries took a long time to build the testing capacity and to implement the testing extensively. The assay design formats were available in the public domain and thereby allowing researchers to replicate them to make diagnostic kits. Consequently, several antigen or antibody-based diagnostic tests were also developed for the diagnosis of COVID-19. However, there were some validation and regulatory challenges while bringing these assays into the market. During the course of the pandemic, it became clear that the countries which implemented testing at an early stage of the pandemic were capable of controlling the spread more effectively than those that implemented them at later stages. As several countries implemented a lockdown for controlling the spread of the virus, it is critical to build the testing capability to meet the extensive need of testing while exiting the lockdown. Testing and isolation of positive cases are the most effective ways of preventing the spread of virus and gradually returning life back to normality.

Comparing Nasopharyngeal Swab and Early Morning Saliva for the Identification of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2).

BackgroundThe ideal severe acute respiratory syndrome coronavirus 2 (SARs-CoV-2) testing method would be accurate and also be patient-performed to reduce exposure to healthcare workers. The aim of this study was to compare patient-performed testing based on a morning saliva sample with the current standard testing method, healthcare worker-collected sampling via a nasopharyngeal swab (NPS).MethodsThis was a prospective single center study which recruited 217 asymptomatic adult male participants in a coronavirus disease 2019 (COVID-19) quarantine center who had tested positive for SARS-CoV-2 8–10 days prior to isolation. Paired NPS and saliva specimens were collected and processed within 5 hours of sample collection. Real time reverse transcription polymerase chain reaction (RT-PCR) targeting Envelope (E) and RNA-dependent RNA polymerase (RdRp) genes was performed and the results were compared.ResultsOverall, 160 of the 217 (74%) participants tested positive for COVID-19 based on saliva, NPS, or both testing methods. The detection rate for SARS-CoV-2 was higher in saliva compared to NPS testing (93.1%, 149/160 vs 52.5%, 84/160, P < .001). The concordance between the 2 tests was 45.6% (virus was detected in both saliva and NPS in 73/160), whereas 47.5% were discordant (87/160 tested positive for 1 whereas negative for the other). The cycle threshold (Ct) values for E and RdRp genes were significantly lower in saliva specimens compared to NP swab specimens.ConclusionsOur findings demonstrate that saliva is a better alternative specimen for detection of SARS-CoV-2. Taking into consideration, the simplicity of specimen collection, shortage of PPE and the transmissibility of the virus, saliva could enable self-collection for an accurate SARS-CoV-2 surveillance testing.

Standardized ethanol extract of Tinospora crispa upregulates pro-inflammatory mediators release in LPS-primed U937 human macrophages through stimulation of MAPK, NF-\u03baB and PI3K-Akt signaling networks

BackgroundImmunomodulatory effects of Tinospora crispa have been investigated due to its traditional use to treat several inflammatory disorders associated to the immune system. The present study reports the underlying mechanisms involved in the stimulation of 80% ethanol extract of T. crispa stems on pro-inflammatory mediators release in lipopolysaccharide (LPS)-primed U937 human macrophages via MyD88-dependent pathways.MethodsRelease of interleukin (IL)-1β and tumor necrosis factor (TNF)-α, and production of prostaglandin E2 (PGE2) were determined by using enzyme-linked immunosorbent assay (ELISA). Immunoblot technique was executed to determine the activation of MAPKs molecules, NF-κB, PI3K-Akt and cyclooxygenase-2 (COX-2) protein. Determination of pro-inflammatory cytokines and COX-2 relative gene expression levels was by performing the real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). A reversed-phase HPLC method was developed and validated to standardize the T. crispa extract and chemical profiling of its secondary metabolites was performed by LC-MS/MS.ResultsQualitative and quantitative analyses of chromatographic data indicated that syringin and magnoflorine were found as the major components of the extract. T. crispa-treatment prompted activation of NF-κB by enhancing IKKα/β and NF-κB (p65) phosphorylation, and degradation of IκBα. The extract upregulated COX-2 protein expression, release of pro-inflammatory mediators and MAPKs (ERK, p38 and JNK) phosphorylation as well as Akt dose-dependently. T. crispa extract also upregulated the upstream signaling adaptor molecules, toll-like receptor 4 (TLR4) and MyD88. T. crispa-treatment also upregulated the pro-inflammatory markers mRNA expression.ConclusionThe results suggested that T. crispa extract stimulated the MyD88-dependent signaling pathways by upregulating the various immune inflammatory related parameters.

Should cyclosporine be useful in renal transplant recipients affected by SARS-CoV-2?

Minimization of immunosuppression and administration of antiretrovirals have been recommended for kidney transplant recipients (KTRs) with coronavirus disease 2019 (COVID‐19). However, outcomes remain poor. Given the likely benefit of cyclosporine because of its antiviral and immunomodulatory effect, we have been using it as a strategy in KTRs diagnosed with severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). We studied 29 kidney transplant recipients (KTRs) who were admitted to our institution with COVID‐19 between March 15and April, 24, 2020. Mycophenolate and/or mammalian target of rapamycin inhibitors (mTORi) were discontinued in all patients. Two therapeutic strategies were compared: Group 1, minimization of calcineurin inhibitors (N = 6); and Group 2, cyclosporine‐based therapy (N = 23), with 15 patients switched from tacrolimus. Hydroxychloroquine was considered in both strategies but antivirals in none. Six patients died after respiratory distress (20.6%). Five required mechanical ventilation (17.2%), and 3 could be weaned. Nineteen patients had an uneventful recovery (65.5%). In group 1, 3 of 6 patients died (50%) and 1 of 6 required invasive mechanical ventilation (16.7%). In group 2, 3 of 23 patients died (12.5%). Renal function did not deteriorate and signs of rejection were not observed in any patient on the second treatment regime. In conclusion, immunosuppressant treatment based on cyclosporine could be safe and effective for KTRs diagnosed with COVID‐19.

Modern Strategy and Biological Fate of Acute Pneumonia

This is a retrospective review of consecutive patients admitted with diagnosis of COVID-19 confirmed by Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR). Clinical, laboratory, radiology and treatment outcome data were obtained by reviewing electronic medical record.

Efficacy of chloroquine versus lopinavir/ritonavir in mild/general COVID-19 infection: a prospective, open-label, multicenter, randomized controlled clinical study

BackgroundThe outbreak of COVID-19 (caused by SARS-Cov-2) is very serious, and no effective antiviral treatment has yet been confirmed. The adage “old drug, new trick” in this context may suggest the important therapeutic potential of existing drugs. We found that the lopinavir/ritonavir treatment recommended in the fifth edition of the Treatment Plan of China can only help to improve a minority of throat-swab nucleic-acid results (3/15) in hospitals. Our previous use of chloroquine to treat patients with COVID-19 infection showed an improvement in more throat-swab nucleic-acid results (5/10) than the use of lopinavir/ritonavir.Methods/designThis is a prospective, open-label, randomized controlled, multicenter clinical study. The study consists of three phases: a screening period, a treatment period of no more than 10 days, and a follow-up period for each participant. Participants with COVID-19 infection who are eligible for selection for the study will be randomly allocated to the trial group or the control group. The control group will be given lopinavir/ritonavir treatment for no more than 10 days. The trial group will be given chloroquine phosphate treatment for no more than 10 days. The primary outcome is the clinical recovery time at no more than 28 days after the completion of therapy and follow-up. The secondary outcomes include the rate of treatment success after the completion of therapy and follow-up, the time of treatment success after no more than 28 days, the rate of serious adverse events during the completion of therapy and follow-up, and the time to return to normal temperature (calculated from the onset of illness) during the completion of therapy and follow-up. Comparisons will be performed using two-sided tests with a statistical significance level of 5%.DiscussionThis experiment should reveal the efficacy and safety of using chloroquine versus lopinavir/ritonavir for patients with mild/general COVID-19 infection. If the new treatment including chloroquine shows a higher rate of throat-swab SARS-CoV-2 real-time fluorescent reverse transcription polymerase chain reaction (RT-PCR) negativity and is safe, it could be tested as a future COVID-19 treatment.Trial registrationChinese Clinical Trial Registry, ID: ChiCTR2000029741. Registered on 11 February 2020.

Detection of COVID-19 Infection from Routine Blood Exams with Machine Learning: A Feasibility Study.

The COVID-19 pandemia due to the SARS-CoV-2 coronavirus, in its first 4 months since its outbreak, has to date reached more than 200 countries worldwide with more than 2 million confirmed cases (probably a much higher number of infected), and almost 200,000 deaths. Amplification of viral RNA by (real time) reverse transcription polymerase chain reaction (rRT-PCR) is the current gold standard test for confirmation of infection, although it presents known shortcomings: long turnaround times (3-4 hours to generate results), potential shortage of reagents, false-negative rates as large as 15-20%, the need for certified laboratories, expensive equipment and trained personnel. Thus there is a need for alternative, faster, less expensive and more accessible tests. We developed two machine learning classification models using hematochemical values from routine blood exams (namely: white blood cells counts, and the platelets, CRP, AST, ALT, GGT, ALP, LDH plasma levels) drawn from 279 patients who, after being admitted to the San Raffaele Hospital (Milan, Italy) emergency-room with COVID-19 symptoms, were screened with the rRT-PCR test performed on respiratory tract specimens. Of these patients, 177 resulted positive, whereas 102 received a negative response. We have developed two machine learning models, to discriminate between patients who are either positive or negative to the SARS-CoV-2: their accuracy ranges between 82% and 86%, and sensitivity between 92% e 95%, so comparably well with respect to the gold standard. We also developed an interpretable Decision Tree model as a simple decision aid for clinician interpreting blood tests (even off-line) for COVID-19 suspect cases. This study demonstrated the feasibility and clinical soundness of using blood tests analysis and machine learning as an alternative to rRT-PCR for identifying COVID-19 positive patients. This is especially useful in those countries, like developing ones, suffering from shortages of rRT-PCR reagents and specialized laboratories. We made available a Web-based tool for clinical reference and evaluation (This tool is available at https://covid19-blood-ml.herokuapp.com/).

Identification of serum exosomal miR-148a as a novel prognostic biomarker for breast cancer.

Breast cancer (BC) is one of the most common malignanciesD. Li, J. Wang, L.-J. Ma, H.-B. Yang, J.-F. Jing, M.-M. Jia, X.-J. Zhang, F. Guo, J.-N. Gao affecting females. Aberrant expression of microRNAs (miRNAs) has been associated with carcinogenesis of BC. The aim of our study was to investigate the correlation of serum exosomal miR-148a expression and the clinical outcome of patients with BC. Quantitative Real Time-Reverse Transcription Polymerase Chain Reaction (qRT-PCR) was applied to evaluate the expression level of serum exosomal miR-148a in patients with BC, patients with benign breast tumors, and healthy controls. Then, the clinical value of serum exosomal miR-148a in BC was evaluated. Serum exosomal miR-148a levels were gradually downregulated from healthy controls patients with benign breast tumors to BC patients. Serum exosomal miR-148a could well distinguish BC patients from healthy volunteers. The expression level of serum exosomal miR-148a in BC patients was significantly upregulated following surgery, while dropped in the cases with disease relapse. A significant association was found between serum exosomal miR-148a levels and the tumor-node-metastasis (TNM) stage, differentiation, and lymph node metastasis in BC. In addition, BC patients with lower expression of serum exosomal miR-148a levels suffered worse overall survival and disease-free survival than those with higher expression of serum exosomal miR-148a levels. Furthermore, serum exosomal miR-148a was an independent risk factor for BC. Our data have demonstrated that serum exosomal miR-148a is significantly reduced in patients with BC and downregulation of serum exosomal miR-148a is closely associated with unfavorable clinical outcome of BC, indicating that serum exosomal miR-148a might serve as a promising diagnostic and prognostic biomarker for BC.

Methylation-sensitive amplification polymorphism analysis reveals the DNA methylation dynamics during capitulum development in Chrysanthemum lavandulifolium

Chrysanthemum lavandulifolium (Fischer ex Trautv.) Makino is a diploid plant belonging to the Asteraceae family, with typical capitula composed of female ray florets and bisexual disc florets. The differentiation and development of these two types of florets have long been important research focuses; however, the potential epigenetic mechanisms governing these processes have not been elucidated. In the present study, methylation-sensitive amplification polymorphism method was used to trace the dynamic changes of DNA methylation during capitulum development in C. lavandulifolium. DNA methylation patterns and levels were detected in the whole capitula during seven developmental stages, and the obtained results revealed that DNA demethylation was dominant during this process. In addition, DNA methylation patterns and levels showed significant differences between ray and disc florets. Moreover, the expression patterns of candidate genes potentially involved in the development processes of two types of florets were analyzed by real-time quantitative reverse transcription polymerase chain reaction, and correlation analysis indicated that the expression levels of ClPI, ClAG2, ClSEP1, ClCYC2c, ClCYC2d, and ClCYC2e were highly correlated with DNA methylation levels. These results indicate that DNA methylation may be involved in the differentiation and development of ray and disc florets. This study provides epigenetic insights into the capitulum development in C. lavandulifolium.

SARS-CoV-2 RNA detection in the air and on surfaces in the COVID-19 ward of a hospital in Milan, Italy

The COVID-19 outbreak has rapidly progressed worldwide finding the health system, scientists and society unprepared to face a little-known, fast spreading, and extremely deadly virus. Italy is one of the countries hardest hit by the pandemic, resulting in healthcare facilities bearing heavy burdens and severe restrictive measures. Despite efforts to clarify the virus transmission, especially in indoor scenarios, several aspects of SARS-CoV-2 spread are still rudimentary. This study evaluated the contamination of the air and surfaces by SARS-CoV-2 RNA in the COVID-19 isolation ward of a hospital in Milan, Italy. A total of 42 air and surface samples were collected inside five different zones of the ward including contaminated (COVID-19 patients' area), semi-contaminated (undressing room), and clean areas. SARS-CoV-2 RNA detection was performed using real time reverse transcription polymerase chain reaction. Overall, 24.3% of swab samples were positive, but none of these were collected in the clean area. Thus, the positivity rate was higher in contaminated (35.0%) and semi-contaminated (50.0%) areas than in clean areas (0.0%; P<0.05). The most contaminated surfaces were hand sanitizer dispensers (100.0%), medical equipment (50.0%), medical equipment touch screens (50.0%), shelves for medical equipment (40.0%), bedrails (33.3%), and door handles (25.0%). All the air samples collected from the contaminated area, namely the intensive care unit and corridor, were positive while viral RNA was not detected in either semi-contaminated or clean areas. These results showed that environmental contamination did not involve clean areas, but the results also support the need for strict disinfection, hand hygiene and protective measures for healthcare workers as well as the need for airborne isolation precautions.

Renal profile of COVID-19 infected patients admitted in a tertiary care hospital in Western Rajasthan

Background: A recent cluster of pneumonia cases all over the world including India, is caused by a novel beta coronavirus, the 2019 novel coronavirus (2019-nCoV). The objective of this study was to observe the effect on renal profiles at admission of these COVID-19 infected patients.Methods: All the patients were admitted to Dr. S. N. Medical College and Hospital, Jodhpur, a tertiary care hospital of Western Rajasthan and were tested for COVID-19 by real time reverse transcription polymerase chain reaction assay of 2019-nCoVRNA. All data in relation to patients including blood renal profile were collected from data collection form from electronic medical records and history given by COVID-19 infected patients admitted.Results: Out of total 300 COVID-19 patients, majority of patient were male i.e. 210 (70%) and majority of the patients 70% were below 60 years of age. The underlying diseases were present in 98 (32.67%) patients. Cough was the most common symptom (80%) followed by fever (72%) in out of 180 (60%) symptomatic patients. 64 (21.3%) patients had deranged renal function tests with abnormal blood urea nitrogen and serum creatinine. Overall, 56 patients (18.6%) without chronic kidney disease showed mild increase of BUN or serum creatinine. 35 patients (11.6%) had high blood urea nitrogen and 18 patients (6%) had raised creatinine.Conclusions: In setting of COVID-19 infected patient’s presentation, renal profile remained essentially normal with deranged RFT seen in patients with comorbidities. Nevertheless, the renal function of patients with COVID-19 needs to be monitored regularly, especially in patients with elevated plasma creatinine.