A pilot study on the expression of circadian clock genes in the alveolar bone of mice with periodontitis.

This study aimed to investigate the expression of circadian clock genes in mouse alveolar bone, and the possible reasons for these changes. Fifty C57 mice were orally inoculated with P. gingivalis, establishing a model of periodontitis using healthy mice as controls. The alveolar bone of both groups was taken for micro-computed tomography scanning to measure the amount of attachment loss, and the relative expression of mRNA in each clock gene and periodontitis related inflammatory factor was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). After the establishment of the mouse model, the height of alveolar bone in the periodontitis group was significantly lower than that in the normal group (p < 0.05). The relative transcriptional level of Bmal1, Per2, and Cry1 mRNA was in the circadian rhythm in the normal group (p ≤ 0.05), while in the periodontitis group, its circadian rhythm disappeared and the transcriptional level characteristics were changed. Interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α), and interferon (IFN-γ) mRNA transcriptional level were elevated in the periodontitis group compared to the normal group. In conclusion, the mRNA transcriptional level of Bmal1, Per2, and Cry1 in alveolar bone of normal mice has circadian rhythm, but the rhythm disappears under the condition of periodontitis, and the cause of its occurrence may be related to inflammatory cytokines.

The expression profile of 79 genes from 107 viruses revealed new insights into disease susceptibility in rats, mice, and muskrats.

The coronavirus disease 2019 (COVID-19), which is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has rapidly spread over the world, resulting in a global severe pneumonia pandemic. Both the cell receptor angiotensin-converting enzyme 2 (ACE2) and the breakdown of S protein by transmembrane serine protease 2 (TMPRSS2) are required by SARS-CoV-2 to enter the host cells. Similarly, the expression level of viral receptor genes in various organs determines the likelihood of viral infection. Several animal species have been found to be infected by the SARS-CoV-2, such as minks, posing an enormous threat to humans. Because the mice and rats were closely related to human and the fact that rats and mice have a risk of infection by SARS-CoV-2 with specific variants, we investigated the expression patterns of 79 receptor genes from 107 viruses, including SARS-CoV-2, in 14 organs of the rat and mouse, and 5 organs of the muskrat, to find the most likely host organs to become infected with certain viruses. The findings of this study are anticipated to aid in prevention of zoonotic infections spread by rats, mice, muskrats, and other rodents.

Based on CT radiomics model for predicting the response to first-line chemotherapy of diffuse large B-cell lymphoma / 中华肿瘤杂志

Objective: To investigate the potential value of CT Radiomics model in predicting the response to first-line chemotherapy in diffuse large B-cell lymphoma (DLBCL). Methods: Pre-treatment CT images and clinical data of DLBCL patients treated at Shanxi Cancer Hospital from January 2013 to May 2018 were retrospectively analyzed and divided into refractory patients (73 cases) and non-refractory patients (57 cases) according to the Lugano 2014 efficacy evaluation criteria. The least absolute shrinkage and selection operator (LASSO) regression algorithm, univariate and multivariate logistic regression analyses were used to screen out clinical factors and CT radiomics features associated with efficacy response, followed by radiomics model and nomogram model. Receiver operating characteristic (ROC) curve, calibration curve and clinical decision curve were used to evaluate the models in terms of the diagnostic efficacy, calibration and clinical value in predicting chemotherapy response. Results: Based on pre-chemotherapy CT images, 850 CT texture features were extracted from each patient, and 6 features highly correlated with the first-line chemotherapy effect of DLBCL were selected, including 1 first order feature, 1 gray level co-occurence matrix, 3 grey level dependence matrix, 1 neighboring grey tone difference matrix. Then, the corresponding radiomics model was established, whose ROC curves showed AUC values of 0.82 (95% CI: 0.76-0.89) and 0.73 (95% CI: 0.60-0.86) in the training and validation groups, respectively. The nomogram model, built by combining validated clinical factors (Ann Arbor stage, serum LDH level) and CT radiomics features, showed an AUC of 0.95 (95% CI: 0.90-0.99) and 0.91 (95% CI: 0.82-1.00) in the training group and the validation group, respectively, with significantly better diagnostic efficacy than that of the radiomics model. In addition, the calibration curve and clinical decision curve showed that the nomogram model had good consistency and high clinical value in the assessment of DLBCL efficacy. Conclusion: The nomogram model based on clinical factors and radiomics features shows potential clinical value in predicting the response to first-line chemotherapy of DLBCL patients.

Effect of muscone on anti-apoptotic ability of muskrat prostate primary cells.

Muskrat is a small fur animal with a pair of scent glands that can secrete muskrat musk during breeding season. The consensus is muskrat musk functions as a pheromone, but we hypothesized it has a broader role. In previous research, we found the presence of muscone in muskrat musk. To study whether the muscone can affect the apoptosis of muskrat prostate, we carried out the following investigations. Primary muskrat prostate cells were cultured and treated with muscone. Then we drew cell proliferation curves by applying the CCK-8 and used TdT-mediated dUTP nick end labeling (TUNEL) to detect apoptosis. Levels of mRNA transcription and protein expression of Bcl-2 as well as Bax were detected by qRT-PCR and the Western blot. Meanwhile, we collected tissue samples of muskrat prostates and froze sections to analyze the fluorescence signal intensity of BCL-2 and BAX via immunofluorescence. Under the treatment of 30 µmol/L muscone, the proliferation rate of the experimental group exceeded that of the control group, and the proportion of cells undergoing apoptosis was lower in the experimental group. The qRT-PCR and Western blot result showed that, in the experimental group, the ratio of Bcl-2 to Bax mRNA transcription levels increased by 2.85 times and their corresponding protein expression ratio increased by 2.37 times (P < 0.05). Immunofluorescence results were consistent with the cell experiment's results. The fluorescence signal intensity of BCL-2 was higher in the breeding season than nonbreeding season but vice versa for BAX. Based on these results, we speculate that the muscone could regulates prostate development by inhibiting apoptosis.

The role of a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type receptor modulators in object recognition memory reconsolidation.

OBJECTIVE: Numerous studies suggest that the a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type (AMPA) receptor appears to play a central role in mediating brain functions, such as learning and memory. Trafficking of this receptor is related to different long-term memory processes. This study explores the role of two AMPA receptor (AMPAR) modulators in object recognition memory (ORM) reconsolidation. METHODS: First, the effects of immediate administration of each drug after memory reactivation were investigated and compared. Then, this drug's efficient time window and its effects without memory reactivation were investigated. RESULTS: Immediate CX546 administration after reactivation did not affect ORM reconsolidation. In contrast, administration of 10-mg/kg NBQX significantly impaired ORM reconsolidation within a 6-h time window. Importantly, the observed effects were not attributed to the exploratory behavior or locomotor activity of mice. CONCLUSION: These findings provide new evidence that the AMPA receptor plays an important role in the reconsolidation phase of ORM.

Cdc13 is predominant over Stn1 and Ten1 in preventing chromosome end fusions.

Telomeres define the natural ends of eukaryotic chromosomes and are crucial for chromosomal stability. The budding yeast Cdc13, Stn1 and Ten1 proteins form a heterotrimeric complex, and the inactivation of any of its subunits leads to a uniformly lethal phenotype due to telomere deprotection. Although Cdc13, Stn1 and Ten1 seem to belong to an epistasis group, it remains unclear whether they function differently in telomere protection. Here, we employed the single-linear-chromosome yeast SY14, and surprisingly found that the deletion of CDC13 leads to telomere erosion and intrachromosome end-to-end fusion, which depends on Rad52 but not Yku. Interestingly, the emergence frequency of survivors in the SY14 cdc13Δ mutant was ~29 fold higher than that in either the stn1Δ or ten1Δ mutant, demonstrating a predominant role of Cdc13 in inhibiting telomere fusion. Chromosomal fusion readily occurred in the telomerase-null SY14 strain, further verifying the default role of intact telomeres in inhibiting chromosome fusion.

A mannose receptor is involved in the anti-Vibrio defense of red swamp crayfish.

Mannose receptor (MR), a member of pattern-recognition receptors (PRRs), is the first MR family member to be discovered that plays a critical role in immunity. The function of MRs has been reported in mammals and teleosts while none in invertebrates. In the present study, we identified a MR-like gene (designated as PcMR) from red swamp crayfish, Procambarus clarkii. The PcMR cDNA is 6848 bp long with a 6288 bp open reading frame that encodes a polypeptide with 2095 amino acid residues. PcMR transcripts were mainly detected in hepatopancreas and hemocytes, and upregulated by Vibrio anguillarum challenge. The PcMR protein contained 14 C-type lectin domains (CTLDs) and they were divided into four fragments (CTLD 1-3, CTLD 4-6, CTLD 7-10, CTLD 11-14). The four recombinant proteins encoded by the four fragments were all expressed and purified. Microorganism-binding and sugar-binding assay showed that CTLD 1-3, CTLD 4-6, CTLD 7-10, CTLD 11-14 could bind to a variety of bacteria, as well as glycoconjugates on the bacterial surface. Moreover, they agglutinated bacteria in a calcium-dependent manner. Bacteria clearance experiment manifested that the mixed proteins facilitated the clearance of injected bacteria in crayfish. PcMR silencing by siRNA interference impaired the bacterial clearance ability. These results suggest PcMR is involved in the antibacterial defense of crayfish, and this study will help us better understand the functions of invertebrate MRs.

Identification of a C-type lectin possessing both antibacterial and antiviral activities from red swamp crayfish.

C-type lectins are important immune molecules that participate in crustacean defense response. The present work reports a novel C-type lectin (PcLec6) from the red swamp crayfish Procambarus clarkii. PcLec6 encodes a single-peptide protein of 385 amino acids, which include a C-type lectin domain (CTLD) and a serine-rich region. PcLec6 expression in lymph organ and gills was up-regulated after bacterial challenge by Vibrio alginolyticus or white spot syndrome virus (WSSV). Recombinant full-length PcLec6 or its CTLD proteins were used for the functional analyses. Results showed that these two proteins had the capacity to bind to carbohydrates and bacteria. Both the full-length PcLec6 and CTLD facilitated the bacterial clearance, but only full-length PcLec6 protected crayfish from WSSV infection. Furthermore, PcLec6 regulated the expression of ALF genes. These results suggest that PcLec6 is involved in the innate immune response of crayfish against both bacterial and viral pathogens.

Comparison of double intravenous vasopressor automated system using nexfin versus manual vasopressor bolus administration for maintenance of haemodynamic stability during spinal anaesthesia for caesarean delivery: A randomised double-blind controlled trial.

BACKGROUND: Hypotension is a common side effect of spinal anaesthesia during caesarean delivery and is associated with maternal and foetal adverse effects. We developed an updated double intravenous vasopressor automated (DIVA) system that administers phenylephrine or ephedrine based on continuous noninvasive haemodynamic monitoring using the Nexfin device. OBJECTIVE: The aim of our present study is to compare the performance and reliability of the DIVA system against Manual Vasopressor Bolus administration. DESIGN: A randomised, double-blind controlled trial. SETTING: Single-centre, KK Women's and Children's Hospital, Singapore. PATIENTS: Two hundred and thirty-six healthy women undergoing elective caesarean delivery under spinal anaesthesia. MAIN OUTCOME MEASURES: The primary outcome was the incidence of maternal hypotension. The secondary outcome measures were reactive hypertension, total vasopressor requirement and maternal and neonatal outcomes. RESULTS: The DIVA group had a significantly lower incidence of maternal hypotension, with 39.3% (46 of 117) patients having any SBP reading less than 80% of baseline compared with 57.5% (65 of 113) in the manual vasopressor bolus group (P = 0.008). The DIVA group also had fewer hypotensive episodes than the manual vasopressor bolus group (4.67 versus 7.77%; P < 0.0001). There was no difference in the incidence of reactive hypertension or the total vasopressor requirement. The DIVA group had less wobble in system performance. Maternal and neonatal outcomes were similar. CONCLUSION: The DIVA system achieved better control of maternal blood pressure after spinal anaesthesia than manual vasopressor bolus administration. TRIAL REGISTRATION: Clinicaltrials.gov identifier: NCT02277730.

Effect of a LECT2 on the immune response of peritoneal lecukocytes against Vibrio anguillarum in roughskin sculpin.

Leukocyte cell-derived chemotaxin 2 (LECT2) is a multi-functional protein that is mainly synthesized by the liver. However, its role in roughskin scalping is less known. Here, we cloned a leukocyte cell-derived chemotaxin 2 (TfLECT2) genes in the liver of roughskin scalping, Trachidermus fasciatus, and studied its possible role involved in the immune response against Vibrio anguillarum (V. anguillarum) of peritoneal lecukocytes under in vivo conditions. The cDNA sequence of TfLECT2 is 566 bp in size. Its deduced amino acid (aa) sequence comprises 151 residues, of which the first 16 residues form a putative signal peptide and 101 residues compose a typical peptidase M23 domain in the C-terminal region. The domain structure is conserved in all LECT2 proteins, which suggests a close phylogenetic relationship between TfLECT2 and LECT2 in other fish species. Real-time quantitative PCR analysis revealed that TfLECT2 gene expression was dramatically increased in liver after V. anguillarum stimulation. Subsequently, TfLECT2 was prokaryotic expressed and purified to prepare anti-TfLECT2 antibody. After V. anguillarum challenge, leukocytes recruitment and LECT2 levels in peritoneal exudates were increased, and positively correlated with each other. Moreover, recombinant TfLECT2 administration significantly improved immune responses after infection, principally in stimulating the recruitment, phagocytosis and respiratory burst of leukocytes at the site of infection; however, anti-TfLECT2 treatment neutralized these abilities. Therefore, TfLECT2 may trigger the early immune events of peritoneal leukocytes and it will be useful to induce innate immune response of fish.

Characterization of a double WAP domain-containing protein from the red swamp crayfish Procambarus clarkii.

Crustaceans express multiple whey acidic protein (WAP) domain containing proteins which are components of host immunity. In the present study, a new double WAP domain containing protein was identified from red swamp crayfish Procambarus clarkii, designated Pc-DWD. The ORF is 387 bp, encoding 128 amino acids consisting of signal peptide of 18 residues, and two tandem WAP domains of 38 and 44 residues. Multiple alignment indicates the presence of conserved motifs in both WAP domains, and phylogenetic analysis shows that Pc-DWD is a new member of the type-IV crustin family. Pc-DWD transcripts were found most abundantly in hemocytes, gills, intestine and heart, and induced by Vibrio anguillarum, Staphylococcus aureus and white spot syndrome virus challenge. RNAi knockdown of Pc-DWD expression led to increased expression of white spot syndrome virus genes and increased crayfish mortality after virus infection. Recombinant Pc-DWD exhibited strong protease inhibitory activity towards commercial subtilicin A and protease K. Pc-DWD inhibited the crude proteases from V. anguillarum and S. aureus cultures and from the crayfish tissue extracts. We infer that Pc-DWD acts in crayfish bacterial and viral immunity.

HcToll3 was involved in anti-Vibrio defense in freshwater pearl mussel, Hyriopsis cumingii.

Toll-like receptors (TLRs) play an important role in the activation of innate immune response but their functions in bivalves remain largely unknown. In this study, we identified a TLR from the freshwater pearl mussel Hyriopsis cumingii (HcToll3) and investigated its functions in immunity. The full-length cDNA of HcToll3 is 3852 bp and includes an open reading frame (ORF) of 3228 bp that encodes a polypeptide of 1075 amino acids. The predicted HcToll3 protein shares similar structural characteristics with other known Toll family proteins. Quantitative real-time PCR analysis revealed that HcToll3 mRNA is broadly expressed in all of the examined tissues; its transcript level was significantly up-regulated by challenge with gram-negative bacteria Vibrio parahaemolyticus or lipopolysaccharide, but not gram-positive Staphylococcus aureus or peptidoglycan. RNA interference by siRNA results showed that HcToll3 regulated expression of whey acidic protein (HcWAP) and lysozymes (HcLyso1 and HcLyso2) in vivo and knockdown of HcToll3 suppressed the elimination of V. parahaemolyticus. These findings suggest that HcToll3 might be involved in anti-Vibrio defense in H. cumingii.

Global trends of municipal solid waste research from 1997 to 2014 using bibliometric analysis.

UNLABELLED: This study uses a bibliometric approach to identify global trends related to the municipal solid waste (MSW). It applies related literature in the Science Citation Index Expanded (SCI-EXPANDED), Social Sciences Citation Index (SSCI), Conference Proceedings Citation Index - Science (CPCI-S) and Conference Proceedings Citation Index - Social Science & Humanities (CPCI-SSH), retrieved from the ISI Web of Science. The data used covers the period from 1997 to 2014. Analyzed aspects included document type, and publication output as well as distribution of journals, subject category, countries, institutions, title-words, author keywords, and keywords plus. An evaluating indicator, citation score, was applied to characterize the MSW publications. The research outputs of MSW had steadily increased in the field of energy fuels, engineering chemical and biotechnology applied microbiology, especially environmental sciences and engineering environmental. The predominance of Chinese institutions in terms of article count and a predominance of industrialized countries' institutions in terms of citation score were compared. Finally, author keywords, words in title, and keywords plus were analyzed to provide research emphasis, with the developing trends and recent hotspots provided. IMPLICATIONS: A systematic overview of scientific literature dealing with municipal solid waste (MSW) is provided by a bibliometric analysis. The analysis of author keywords helps in drawing the research trends in a special perspective. Research studies on food waste, life cycle assessment (LCA), and renewable energy tend to be a new research focus in the area of MSW. The conclusions could provide a reference to the decision making and policy of MSW management for the government to some extent.

Knowledge, attitude, and practice of dry eye treatment by institutional Chinese physicians in Singapore.

Dry eye is a common health problem worldwide, causing significant discomfort and inconvenience to sufferers. The conventional treatment of dry eye via topical administration of eye drops is deemed palliative and unsatisfactory to many. Traditional Chinese medicine (TCM) has shown some promise in dry eye treatment; however, the extent of its use and acceptance is uncertain. We evaluated the knowledge, attitude, and practice of institutional TCM practitioners in the treatment of dry eye in Singapore. A questionnaire was generated to address the study aims and sent to TCM practitioners listed in the Singapore TCM practitioners' board database. About three quarters of respondents thought that dry eye was not severe enough to be a public health burden but most thought that TCM was effective in the treatment of dry eye. Acupuncture and herbal medicine were most commonly used TCM modalities in dry eye treatment, and a single TCM treatment session would be charged S$20-50 by the practitioner. The majority of surveyed institutional TCM practitioners in Singapore believe that TCM is relevant in dry eye treatment. Public awareness should be raised regarding the availability of TCM as alternative medicine for dry eye.

Cost of dry eye treatment in an Asian clinic setting.

OBJECTIVES: To estimate the cost and patterns of expenditure of dry eye treatment. METHODOLOGY: We retrieved data on the type and cost of dry eye treatment in Singapore National Eye Centre from pharmacy and clinic inventory databases over a 2 year period (2008-2009) retrospectively. According to the type of treatment, data were sorted into 7 groups; meibomien gland disease (MGD) treatment, preservative free lubricant eye drops, preserved lubricant eye drops, lubricant ointments and gels, cyclosporine eye drops, oral supplements and non-pharmacological treatments/procedures. Each recorded entry was considered as one patient episode (PE). Comparisons in each group between two years were carried out using Pearson Chi-Square test. Significance level was set at alpha  =  0.05. RESULTS: Cost data from 54,052 patients were available for analysis. Total number of recorded PEs was 132,758. Total annual expenditure on dry eye treatment for year 2008 and 2009 were US$1,509,372.20 and US$1,520,797.80 respectively. Total expenditure per PE in year 2008 and 2009 were US$22.11 and US$23.59 respectively. From 2008 to 2009, there was a 0.8% increase in total annual expenditure and 6.69% increase in expenditure per PE. Pharmacological treatment attributes to 99.2% of the total expenditure with lubricants accounting for 79.3% of the total pharmacological treatment expenditure. Total number of units purchased in preservative free lubricants, cyclosporine eye drops and MGD therapy have increased significantly (p<0.001) whereas number of units purchased in preserved lubricants and ointments/gels have reduced significantly (p<0.001) from 2008 to 2009. CONCLUSION: Dry eye imposes a significant direct burden to health care expenditure even without considering indirect costs. Health care planners should be aware that these direct costs appear to increase over the time and more so for particular types of medications. Given the limitations of socio-economic data, true societal costs of Dry eye syndrome are likely to be much higher than estimated.

Optimization of human corneal endothelial cells for culture: the removal of corneal stromal fibroblast contamination using magnetic cell separation.

The culture of human corneal endothelial cells (CECs) is critical for the development of suitable graft alternative on biodegradable material, specifically for endothelial keratoplasty, which can potentially alleviate the global shortage of transplant-grade donor corneas available. However, the propagation of slow proliferative CECs in vitro can be hindered by rapid growing stromal corneal fibroblasts (CSFs) that may be coisolated in some cases. The purpose of this study was to evaluate a strategy using magnetic cell separation (MACS) technique to deplete the contaminating CSFs from CEC cultures using antifibroblast magnetic microbeads. Separated "labeled" and "flow-through" cell fractions were collected separately, cultured, and morphologically assessed. Cells from the "flow-through" fraction displayed compact polygonal morphology and expressed Na(+)/K(+)ATPase indicative of corneal endothelial cells, whilst cells from the "labeled" fraction were mostly elongated and fibroblastic. A separation efficacy of 96.88% was observed. Hence, MACS technique can be useful in the depletion of contaminating CSFs from within a culture of CECs.

[A two-dimensional reference map of mouse ovary proteins].

OBJECTIVE: To perform a preliminary proteomic analysis of mouse ovaries and to study the protein's function in mouse ovary. METHODS: The two-dimensional gel electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) were used to analyze mouse ovarian proteome. A 12.5% sodium dodecyl sulfate (SDS) reference gel was generated by immobilized pH gradient isoelectric focusing of mouse ovary proteins in a non-linear gradient (pH 3-10). And GRP78 was selected to perform with immunohistochemistry within mouse ovaries. RESULTS: Based on peptide mass fingerprinting, 52 proteins were identified and classified into seven functional groups: Cell/organism defense and antioxidant, cell signaling/communications proteins, cell structure/motility proteins, metabolism proteins, RNA synthesis processing, protein synthesis and processing, and unclassified proteins. The immunoreactivity of GRP78 was detected in GCs in the follicular, and with during GCs Luteinizing in the menstrual cycle, the protein expression (brown) increased continually and came to a head when ovulation happened. CONCLUSION: This work provides a first step toward the establishment of a systematic ovary protein database and stands as a valuable resource for molecular analyses of normal and pathologic conditions affecting mouse ovaries.