Effects of Muscone on Cartilage Morphology and Matrix Metalloproteinases-3 (MMP-3h) and Matrix Metalloproteinases-9 (MMP-9) Expression in Rheumatoid Arthritis Rat Models

Aim: To discuss Muscone treatment in Rheumatoid Arthritis Rat Models and relative mechanisms. Materials and methods: Dividing 36 rats as 4 groups as Normal, Model, DMSO and Muscone groups (n = 9). Rats of Model, DMSO and Muscone groups were made Rheumatoid Arthritis model. Muscone group were treated with 2 mg/kg Muscone after modeling. HE staining and Masson staining were used to observe the morphological changes of cartilage tissue, measuring MMP-3 and MMP-9 expression by RT-PCR, Western Blotting (WB) and Immunohistochemistry (IHC). Results: Compared with Model group, the pathological changes of Muscone group was significantly improved and average optical density of collagen fibers was significantly depressed (P < 0.001, respectively) via MMP-3 and MMP-9 proteins significantly depressing (P < 0.001, respectively). Conclusion: Muscone improved Rheumatoid Arthritis by depressing MMP-3 and MMP-9 proteins in vivo study.

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice

The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

Detection of Polioviruses in Sewage Using Cell Culture and Molecular Methods

The work presented here demonstrates the utility of a two-step algorithm for environmental poliovirus surveillance based on: preselection of sewage samples tested for the presence of enteroviral genetic material-RT-PCR assay and detection of infectious viruses by cell culture technique (L20B for polioviruses and RD for polio and other non-polio enteroviruses). RD and L20B cell lines were tested to determine their sensitivity for isolation of viruses from environmental samples (sewage). Finally, we wanted to determine if sewage concentration affects the results obtained for RT-PCR and cell cultures.

miR-653 Induces Bone Marrow Mesenchymal Stem Cells Proliferation via Activating Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma

This study intends to assess miR-653’s expression in MSCs and OSCC and discuss molecular biological mechanism of changes of EMT in MSCs through activating miR-653 in OSCC. miR-653 expression in MSCs and OSCC was detected. si-miR-653 was transfected into MSCs followed by analysis of cell proliferation by CCK-8 and clone formation assay, cell apoptosis and cycle by FCM, and the changes of transcription factor as ZEB1 and Snail by qRT-PCR. miR-653 expression in OSCC cell was up-regulated significantly from the result of q-RT-PCR detection. The proliferation of MSCs induced by miR-653 was restrained and apoptotic rate was increased after treatment with si-miR-653 along with stagnated cycle of G1/G0 staging cell. The expression of transcription factor of EMT type as ZEB1 and Snail was elevated significantly after intervention using si-miR-653. In conclusion, the proliferation of OSCC could be induced by MSCs through activation with miR-653 which might be through regulation of EMT process.

Effect of miR-184 and miR-205 on the tumorigenesis of conjunctival mucosa associated lymphoid tissue lymphoma through regulating RasL10B and TNFAIP8

AIM: To explore the effect of miR-184 and miR-205 on the proliferation and metastasis of conjunctival mucosa associated lymphoid tissue (MALT) lymphoma. METHODS: Tissue of tumor and adjacent normal control from 5 patients with conjunctival MALT was included. RPMI8226 cell line was selected to verify the effect of miRNAs in B cells. The function of microRNA on the RPMI8226 cell apoptosis, migration and invasion was evaluated by apoptosis assay and Transwell assay. The mRNA and protein expression were examined by quantitative RT-PCR and Western blotting. The effect of microRNA on regulation of downstream gene expression was evaluated by luciferase report assay. RESULTS: A decreased level of miR-184 and miR-205 was observed in MALT lymphoma tissue. Exogenous miR-184 and miR-205 analogues promoted apoptosis, and inhibited the survival, migration, and invasion of RPMI8226 cells. miR-184 and miR-205 inhibitor reversed the process. The RNA and protein level of RasL10B and TNFAIP8 were downregulated in MALT lymphoma tissue. The exogenous of miR-184 and miR-205 promoted the expression of RasL10B and TNFAIP8. Meanwhile, inhibition of miR-184 and miR-205 repressed the expression of target gene, RasL10B and TNFAIP8. CONCLUSION: miR-184 and miR-205 suppresses the tumorigenesis of conjunctival MALT lymphoma through regulating RasL10B and TNFAIP8.

Postmortem brain 7T MRI with minimally invasive pathological correlation in deceased COVID-19 subjects

Background Brain abnormalities are a concern in COVID-19, so we used minimally invasive autopsy (MIA) to investigate it, consisting of brain 7T MR and CT images and tissue sampling via transethmoidal route with at least three fragments: the first one for reverse transcription polymerase chain reaction (RT-PCR) analysis and the remaining fixed and stained with hematoxylin and eosin. Two mouse monoclonal anti-coronavirus (SARS-CoV-2) antibodies were employed in immunohistochemical (IHC) reactions. Results Seven deceased COVID-19 patients underwent MIA with brain MR and CT images, six of them with tissue sampling. Imaging findings included infarcts, punctate brain hemorrhagic foci, subarachnoid hemorrhage and signal abnormalities in the splenium, basal ganglia, white matter, hippocampi and posterior cortico-subcortical. Punctate brain hemorrhage was the most common finding (three out of seven cases). Brain histological analysis revealed reactive gliosis, congestion, cortical neuron eosinophilic degeneration and axonal disruption in all six cases. Other findings included edema (5 cases), discrete perivascular hemorrhages (5), cerebral small vessel disease (3), perivascular hemosiderin deposits (3), Alzheimer type II glia (3), abundant corpora amylacea (3), ischemic foci (1), periventricular encephalitis foci (1), periventricular vascular ectasia (1) and fibrin thrombi (1). SARS-CoV-2 RNA was detected with RT-PCR in 5 out of 5 and IHC in 6 out 6 patients (100%). Conclusions Despite limited sampling, MIA was an effective tool to evaluate underlying pathological brain changes in deceased COVID-19 patients. Imaging findings were varied, and pathological features corroborated signs of hypoxia, alterations related to systemic critically ill and SARS-CoV-2 brain invasion.

Deep Ensemble Learning-Based Models for Diagnosis of COVID-19 from Chest CT Images

Novel coronavirus (COVID-19) has been endangering human health and life since 2019. The timely quarantine, diagnosis, and treatment of infected people are the most necessary and important work. The most widely used method of detecting COVID-19 is real-time polymerase chain reaction (RT-PCR). Along with RT-PCR, computed tomography (CT) has become a vital technique in diagnosing and managing COVID-19 patients. COVID-19 reveals a number of radiological signatures that can be easily recognized through chest CT. These signatures must be analyzed by radiologists. It is, however, an error-prone and time-consuming process. Deep Learning-based methods can be used to perform automatic chest CT analysis, which may shorten the analysis time. The aim of this study is to design a robust and rapid medical recognition system to identify positive cases in chest CT images using three Ensemble Learning-based models. There are several techniques in Deep Learning for developing a detection system. In this paper, we employed Transfer Learning. With this technique, we can apply the knowledge obtained from a pre-trained Convolutional Neural Network (CNN) to a different but related task. In order to ensure the robustness of the proposed system for identifying positive cases in chest CT images, we used two Ensemble Learning methods namely Stacking and Weighted Average Ensemble (WAE) to combine the performances of three fine-tuned Base-Learners (VGG19, ResNet50, and DenseNet201). For Stacking, we explored 2-Levels and 3-Levels Stacking. The three generated Ensemble Learning-based models were trained on two chest CT datasets. A variety of common evaluation measures (accuracy, recall, precision, and F1-score) are used to perform a comparative analysis of each method. The experimental results show that the WAE method provides the most reliable performance, achieving a high recall value which is a desirable outcome in medical applications as it poses a greater risk if a true infected patient is not identified.

Rhinoorbital mucormycosis in COVID-19 pandemic: presentation and course of disease: An observational study

During the second wave of covid 19[SARS- Co V-] pandemic, there is a sudden increase in number of mucormycosis infection cases in India. The present study is an attempt to understand the presentation, course and outcome of rhinoorbital mucormycosis in a group of patients who reported to Ophtalmology and Otorhinolaryngology department of our Govt. District Hospital (secondary referral centre) for enhancing measures for prevention and management. Patients who reported to our Government district hospital with signs or symptoms suggestive of rhino orbital mucormycosis during May-June 2021 were included in the study with consent of ethical committee, patients and patient’s relatives. Total 17 cases were reported and followed. Clinical examination was done for all the patients. History of the presenting complaints and underlying illness with COVID -19 was elicited. Underlying comorbid status was recorded. Patients were followed as all of them were referred to higher centre for further management as per the guidelines issued by directorate medical and health services, rajasthan, Jaipur.13(76.4%) patients were from rural and 4 (23.5%) were from urban area. 11(64.7%) patients had RT-PCR +ve, 6 had RT-PCR _ve, 2 did not have RT-PCR report. 15(88.7%) patients had high blood sugar at presentation mean being 315.7mg%. 9 (52.9%) developed mucormycosis during their treatment for COVID in hospital. 8(47.05%) presented in OPD. 9 patients had treatment with inhalational Owhile 8 patients did not have treatment with O Death rate was high (70.5%) among our patients. Patients who survived (29.4%) had only initial symptoms and signs at presentation therefore could be managed earlier. None of our patient had vaccination for COVID. Our study was done at secondary referral centre, all the previous studies were done at tertiary referral centres; therefore it shows the course of disease mainly among rural population ; most of them presented very late and had poor outcomes. It shows the need of more awareness about COVID and mucormycosis among people especially in rural areas. High blood sugar either due to treatment with steroids or pre existing is a major risk factor for Rhino orbital mucormycosis. Being RT- PCR negative for COVID 19 does not rule out the associated possible complication of Rhino orbital mucormycosis. Early diagnosis and management remains the key factor for managing Rhino orbital mucormycosis.

Detection of Banana Mild Mosaic Virus in Musa In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species

: The banana mild mosaic virus (BanMMV) (Betaflexiviridae, Quinvirinae, unassigned species) is a filamentous virus that infects Musa spp. and has a very wide geographical distribution. The current BanMMV indexing process for an accession requires the testing of no less than four plants cultivated in a greenhouse for at least 6 months and causes a significant delay for the distribution of the germplasm. We evaluated the sensitivity of different protocols for BanMMV detection from in vitro plants to accelerate the testing process. We first used corm tissues from 137 in vitro plants and obtained a diagnostic sensitivity (DSE) of only 61% when testing four plants per accession. After thermotherapy was carried out to eliminate BanMMV infection, the meristem was recovered and further grown in vitro. The same protocol was evaluated in parallel on the corm tissue surrounding the meristem, as a rapid screening to evaluate virus therapy success, and was compared to the results obtained following the standard protocol. The obtained results showed 28% false negatives when conducting testing from corm tissues, making this protocol unsuitable in routine processes. Furthermore, RT-PCR and high-throughput sequencing (HTS) tests were applied on tissues from the base (n = 39) and the leaves (n = 36). For RT-PCR, the average DSE per sample reached 65% from either the base or leaves. HTS was applied on 36 samples and yielded 100% diagnostic specificity (DSP) and 100% DSE, whatever the sampled tissue, allowing the identification of a new Betaflexiviridae species infecting Musa. These results suggest that a reliable diagnostic of BanMMV from in vitro plants using RT-PCR or HTS technologies might represent an efficient alternative for testing after greenhouse cultivation.