CircRNA PVT1 promotes proliferation and chemoresistance of osteosarcoma cells via the miR-24-3p/KLF8 axis.

OBJECTIVE: To investigate the regulatory effect and mechanism of circular RNA PVT1 (circPVT1) in proliferation and chemoresistance of osteosarcoma (OS) cells. METHODS: The expression of circPVT1 in human OS and adjacent normal tissues was detected. The correlation between circPVT1 expression and clinical features of OS was analyzed. The expressions of circPVT1 and miR-24-3p in OS cells resistant to cisplatin, doxorubicin or methotrexate and parental OS cells were detected after cell transfection. CCK-8 and colony formation assay assessed the viability and proliferative ability of OS cells. qRT-PCR and Western blotting measured the expression of KLF8. Dual-luciferase reporter and RNA pull-down assays verified the targeting relationships of circPVT1/miR-24-3p and miR-24-3p/KLF8. RESULTS: CircPVT1 was over-expressed in OS tissues and cells, and correlated with clinical features of OS. Over-expressed circPVT1 reduced the survival of OS patients. CircPVT1 was up-regulated in chemoresistant OS cells compared to their parental cells. CircPVT1 inhibition suppressed the proliferation and chemoresistance of OS cells. MiR-24-3p was under-expressed in OS cells and further down-regulated in chemoresistant cells. CircPVT1 could bind and down-regulate miR-24-3p. MiR-24-3p overexpression inhibited the proliferation and chemoresistance of OS cells. KLF8 was over-expressed in OS cells and further up-regulated in chemoresistant cells. MiR-24-3p negatively regulated the expression of KLF8. CONCLUSION: CircPVT1 mediates proliferation and chemoresistance of OS cells via the miR-24-3p/KLF8 axis. The findings may provide guidance for clinical treatment of OS.

[Pararectus approach for treatment of acetabular both-column fracture combined with translocation of quadrilateral surface].

OBJECTIVE: To study the clinical effect and surgical operating points of pararectus approach for the internal fixation of acetabular both-column fractures with concurrent displaced quadrilateral plate fractures. METHODS: From January 2012 to December 2013, in the Third Affiliated Hospital of Southern Medical University, 15 patients with acetabular both-column fractures and displaced quadrilateral plate fractures were surgically managed through the pararectus approach. There were 11 male and 4 female patients, with an average age of 40 years (from 19 to 61 years). According to Judet-Letournel classification, there were 9 anterior column plus posterior hemitransverse fractures, 6 both-column fractures, 8 cases involving the pelvic fracture. All these fractures were treated through the pararectus approach, in the horizontal position with general anesthesia. The pre-bended plate was placed in interior pelvic ring to fix the anterior wall, anterior column and quadrilateral plate in direct sight. Then, the posterior column was exposed and fixed with antegrade lag screw. Patients were followed up in 4 weeks, 12 weeks, 6 months, 1 year after the operation, and the anteroposterior radiograph of pelvis and the X-ray examination of the fractured hip was performed. RESULTS: All the 15 cases underwent the operation successfully. Postoperative X-ray and CT exams showed excellent and good reduction of anterior column, posterior column and quadrilateral plate, with none surgical complication occurred. According to the Matta radiological evaluation postoperatively, reduction of acetabular fracture was rated as excellent in 9 cases, good in 3 cases and poor in 3 cases. The rate of excellent and good was 12/15. After 8 to 18 months' follow-up (median follow-up time was 14 months), all the patients gained bone union. According to the modified Merle D'Aubigne and Postel scoring system, 9 cases were excellent, 4 were good, and 2 were fair. The rate of excellent and good was 13/15. CONCLUSIONS: Surgical management of acetabular fractures through the pararectus approach can provide adequate exposure of reducing and fixing both-column acetabular fractures with concurrent displaced quadrilateral plate fractures, which has a good effect in clinical application.

Oily fish and raw vegetable consumption can decrease the risk of AQP4-positive neuromyelitis optica spectrum disorders: a Mendelian-randomization study.

Neuromyelitis optica spectrum disorders (NMOSD) are severe inflammatory disorders of the central nervous system targeting aquaporin-4 (AQP4). The risk factors for NMOSD remain to be determined, though they may be related to diet and nutrition. This study aimed to explore the possibility of a causal relationship between specific food intake and AQP4-positive NMOSD risk. The study followed a two-sample Mendelian randomization (MR) design. Genetic instruments and self-reported information on the intake of 29 types of food were obtained from a genome-wide association study (GWAS) on 445,779 UK Biobank participants. A total of 132 individuals with AQP4-positive NMOSD and 784 controls from this GWAS were included in our study. The associations were evaluated using inverse-variance-weighted meta-analysis, weighted-median analysis, and MR-Egger regression. A high consumption of oily fish and raw vegetables was associated with a decreased risk of AQP4-positive NMOSD (odds ratio [OR] = 1.78 × 10-16, 95% confidence interval [CI] = 2.60 × 10-25-1.22 × 10-7, p = 0.001; OR = 5.28 × 10-6, 95% CI = 4.67 × 10-11-0.598, p = 0.041, respectively). The results were consistent in the sensitivity analyses, and no evidence of directional pleiotropy was observed. Our study provides useful implications for the development of AQP4-positive NMOSD prevention strategies. Further research is needed to determine the exact causal relationship and mechanisms underlying the association between specific food intake and AQP4-positive NMOSD.

Acetylcholine receptor binding antibody-associated myasthenia gravis, myocarditis, and rhabdomyolysis induced by tislelizumab in a patient with colon cancer: A case report and literature review.

Despite the intriguing therapeutic prospects offered by immune checkpoint inhibitors (ICIs), immune-related adverse events (irAEs) become an increasingly important safety issue. Herein, we report a patient with locally advanced colorectal cancer (LACRC) who received anti-programmed cell death protein 1 (PD-1) (tislelizumab) therapy, then developed weakness of the limbs and drooping eyelids. He experienced sequential irAEs including severe myasthenia gravis, myocarditis, and rhabdomyolysis. Although many irAEs caused by tislelizumab have been reported, the cooccurrence of severe myasthenia gravis, myocarditis, and rhabdomyolysis caused by tislelizumab has not been described. The patient responded well to methylprednisolone and intravenous immunoglobulin therapy. This case illustrates the severe toxicity caused by ICIs, highlighting the importance of early prevention, early diagnosis, and appropriate management of irAEs. Multidisciplinary discussions should be held to improve the prognosis of patients.

Toxicity of copper, lead, and cadmium on the motility of two marine microalgae Isochrysis galbana and Tetraselmis chui.

Computer assisted movement tracking was used to characterize the motility of two marine microalgae, Isochrysis galbana and Tetraselmis chui, and to investigate the toxicity of Cu, Pb, and Cd on motile percentage, curvilinear velocity, average path velocity, straight line velocity, linearity, straightness, and wobble. Except for motile percentage, all other motility parameters differed significantly between I. galbana and T. chui. Based on relative motile percentage data, the median effective concentration (EC50) of Cu on the motility of I. galbana and T. chui was 31.4 and 1.3 micromol/L, respectively, while for Pb it was 37.8 and 10.9 micromol/L and for Cd it was 121.6 and 37.8 micromol/L, respectively. Compared to I. galbana, T. chui was more sensitive to all tested metals. The toxic effect of the heavy metals on motility exhibited the following decreasing order for both species: Cu > Pb > Cd. Our results indicate that L. galbana and T. chui motility is sensitive to heavy metals and can be used as an indicator for toxicology bioassays.

LPCANet: Classification of Laryngeal Cancer Histopathological Images Using a CNN with Position Attention and Channel Attention Mechanisms.

Laryngeal cancer is one of the most common malignant tumors in otolaryngology, and histopathological image analysis is the gold standard for the diagnosis of laryngeal cancer. However, pathologists have high subjectivity in their diagnoses, which makes it easy to miss diagnoses and misdiagnose. In addition, according to a literature search, there is currently no computer-aided diagnosis (CAD) algorithm that has been applied to the classification of histopathological images of laryngeal cancer. Convolutional neural networks (CNNs) are widely used in various other cancer classification tasks. However, the potential global and channel relationships of images may be ignored, which will affect the feature representation ability. Simultaneously, due to the lack of interpretability, the results are often difficult to accept by pathologists. we propose a laryngeal cancer classification network (LPCANet) based on a CNN and attention mechanisms. First, the original histopathological images are sequentially cropped into patches. Then, the patches are input into the basic ResNet50 to extract the local features. Then, a position attention module and a channel attention module are added in parallel to capture the spatial dependency and the channel dependency, respectively. The two modules produce the fusion feature map to enhance the feature representation and improve network classification performance. Moreover, the fusion feature map is extracted and visually analyzed by the grad-weighted class activation map (Grad_CAM) to provide a certain interpretability for the final results. The three-class classification performance of LPCANet is better than those of the five state-of-the-art classifiers (VGG16, ResNet50, InceptionV3, Xception and DenseNet121) on the two original resolutions (534 * 400 and 1067 * 800). On the 534 * 400 data, LPCANet achieved 73.18% accuracy, 74.04% precision, 73.15% recall, 72.9% F1-score, and 0.8826 AUC. On the 1067 * 800 data, LPCANet achieved 83.15% accuracy, 83.5% precision, 83.1% recall, 83.1% F1-score, and 0.9487 AUC. The results show that LPCANet enhances the feature representation by capturing the global and channel relationships and achieves better classification performance. In addition, the visual analysis of Grad_CAM makes the results interpretable, which makes it easier for the results to be accepted by pathologists and allows the method to become a second tool for auxiliary diagnosis.

Microplastics aggravate the bioaccumulation of three veterinary antibiotics in the thick shell mussel Mytilus coruscus and induce synergistic immunotoxic effects.

Marine bivalves living in pollution-prone coastal areas may be simultaneously coexposed to veterinary antibiotic residuals and microplastics (MPs). However, the synergistic effects of these two types of emergent pollutants have not been fully elucidated in mussel species. Therefore, the immunotoxic effects of the three representative antibiotics, oxytetracycline (OTC, 270 ng/L), florfenicol (FLO, 42 ng/L), and sulfamethoxazole (SMX, 140 ng/L), with and without the copresence of polystyrene MPs (0.26 mg/L, dimeter: 500 nm), were investigated in the thick shell mussel. Our data showed that the immunity was significantly hampered by exposure to the pollutants and MP-antibiotic coexposure induced synergistic immunotoxicity. For instance, compared to those treated with antibiotics (OTC, FLO, and SMX) alone, mussels coexposed to antibiotic and MPs had significantly lower phagocytic rate (further decline by approximately 28.80%, 34.21%, and 11.22%, respectively) and total hemocyte count (further reduced by approximately 37.45%, 61.67%, and 46.32%, respectively). Exposure to the pollutants tested also led to inductions in intracellular reactive oxygen species (ROS), decreases in the F-actin cytoskeleton, declines in the cell viability of hemocytes, and downregulation of cytoskeleton- and immune-related genes. In addition, mussels coexposed to antibiotic-MP accumulated significantly greater amounts of antibiotics, which may partially explain the synergic immunotoxic effect detected. Exposure to pollutants tested also led to suppression in the activity of glutathione-S-transferase (GST) and downregulation of detoxification-related genes whereas induction in the level of lipid peroxidation (indicated by MDA content) in gills, which may facilitate the entry whereas constrain the exclusion of antibiotics and therefore result in an elevation in accumulation of antibiotics.

Nibea coibor growth hormone gene: its phylogenetic significance, microsatellite variation and expression analysis.

The growth hormone (GH) gene has been characterized for a number of fishes and used to establish phylogenetic relationships and as a candidate gene for studies of genetic variation in connection with growth traits. In this study, we report the genomic structure of Nibea coibor GH (designated as ncGH) including its 5'-flanking region, being cloned by homology-cloning and chromosome walking methods. The ncGH gene spans approximately 3.0 kb and consists of six exons and five introns, as found for all cloned teleost GH genes with the exception of carps and catfish. The 5'-flanking region contains consensus sequences for a TATA box, a CRE, a pit-1alpha, a TRE, two HNF-3, a ERE and a GRE. Five microsatellites are identified in the ncGH gene and three of them are polymorphic marker. The open reading frame (ORF) of ncGH is 615 bp in length encoding a polypeptide of 204 amino acids with an estimated molecular mass of 23.04 kDa and theoretical isoelectric point of 7.79. The precursor of ncGH consists of a 17 amino-acid signal peptide and a 187 amino-acid mature peptide. The four Cys residues are located at conserved positions (Cys(69), Cys(177), Cys(194) and Cys(202)), and One possible site for N-glycosylation (Asn-X-Ser/Thr motif) is present at Asn(201). The coding region sequence of ncGH is used to align with the sequences of 18 other species from Percoidei and one species from Anabantoidei using Clustal X. A matrix of 612 bp was used to construct the phylogenetic trees using neighbor-joining and maximum parsimony methods. The phylogenetic trees by two methods are identical in most of the clades with high bootstrap support. Every family all forms independent monophyly on the phylogenetic trees, in the family, the different species also forms the monophyly according to the different genera. The results are also identical to those from morphological data, and demonstrated that the GH gene is very suitable for phylogenetic relationship analysis of Percoidei. To validate the predicted exon/intron boundaries, ncGH cDNA is cloned using RT-PCR, and tissue distributions are investigated using semi-quantitative RT-PCR method. The results indicate that the predicted exon/intron is correct, the ncGH mRNA are mainly expressed in pituitary, and weakly expressed in ovary, brain, liver, gill, intestine, muscle and hear, but not expressed in spleen and kidney.

Flow-injection chemiluminescence method for the determination of naphazoline hydrochloride and oxymetazoline hydrochloride.

A sensitive and simple flow-injection chemiluminescence (FI-CL) method, which was based on the CL intensity generated from the redoxreaction of potassium permanganate (KMnO4)-formaldehyde in vitriol (H2SO4) medium, has been developed, validated and applied for the determination of naphazoline hydrochloride and oxymetazoline hydrochloride. Besides oxidants and sensitizers, the effect of the concentration of H(2)SO(4), KMnO4 and formaldehyde was investigated. Under the optimum conditions, the linear range was 1.0 x 10(-2)-7.0 mg/L for naphazoline hydrochloride and 5.0 x 10(-2)-10.0 mg/L for oxymetazoline hydrochloride. During seven repeated inter-day and intra-day precision tests of 0.1, 1.0 and 10.0 mg/L samples, the relative standard deviations all corresponded to reference values. The detection limit was 8.69 x 10(-3) mg/L for naphazoline hydrochloride and 3.47 x 10(-2) mg/L for oxymetazoline hydrochloride (signal-to-noise ratio < or = 3). This method has been successfully implemented for the determination of naphazoline hydrochloride and oxymetazoline hydrochloride in pharmaceuticals.

Nutritional risk, malnutrition and nutritional support among hospitalized patients in orthopedics/spinal surgery of a Hohhot teaching hospital.

The evolution of nutritional status (the prevalence of nutritional risk, malnutrition, overweight and obesity) and the nutritional support of the hospitalized patients from admission to discharge or over a two-week period in orthopedics/ spinal surgery of a teaching hospital in Hohhot were investigated. 432 patients from two wards of the orthopedics/spinal surgery from Jan to Dec 2013, the traditional spinal surgery and the minimally invasive spinal surgery, were selected and detected in this study. The Nutritional Risk Score 2002 (NRS 2002) was used to determine the patients' nutritional status within 48 h after admission and during their hospitalization. The overall prevalence of nutritional risk, malnutrition, overweight and obesity at admission was 11.6%, 12.7%, 35.9% and 7.41%, respectively. Overall, there were 88.0% of the patients who were at nutritional risk received nutritional support, while 14.1% of non-risk patients received a redundant nutritional support. The overall prevalence of nutritional risk changed from 11.6% at admission to 19.4% upon discharge (p<0.05), and the prevalence of malnutrition changed from 12.7% to 20.6% (p<0.05). The prevalence of overweight and obesity, which changed from 35.9% to 31.0% and from 7.41% to 5.79% respectively, didn't experience statistically significant evolution. NRS 2002 was a feasible nutritional risk screening tool for patients in spinal surgery of orthopedics department. Patients' prevalence of nutritional risk and malnutrition increased significantly in spinal surgery of this hospital. Some inappropriate uses of nutritional support were observed in orthopedics/spinal surgery, and nutritional support guidelines or protocols should be promoted by a professional committee.

Cloning, characterization and expression analysis of interleukin-10 from the zebrafish (Danio rerion).

Cytokines are proteins produced by many different cells of the immune system and play a significant role in initiating and regulating the inflammatory process. In this research, an important cytokine, interleukin-10 (IL-10) gene, has been identified and characterized from zebrafish (Danio rerio) genome database. Zebrafish IL-10 is located within a 2690 bp fragment and contains five exons and four introns, sharing the same organization with mammalian IL-10 genes. An open reading frame of 543 bp was found to encode a putative 180 amino acid protein with a signal peptide of 22 amino acids, which shares 29.7-80.9 % homology with amino acid sequences of other known IL-10. The signature motif of IL-10 is also conserved in zebrafish IL-10. The predicted transcript was finally confirmed by sequencing of cDNA clones. Multi-tissue reverse transcriptase PCR (RT-PCR) was performed to examine the tissue distribution and expression regulation of this gene in seven organs of normal and lipopolysaccharide (LPS) stimulation zebrafish. The results demonstrated that this gene was expressed slightly in normal kidney, gill and gut, no expression was detected in other four tissues. The expression was clearly upregulated after LPS stimulation. Using the ideal zebrafish model, further study of IL-10 characterization and function may provide insight on the understanding of the innate immune system.

Molecular cloning, recombinant expression, and growth-promoting effect of mud carp (Cirrhinus molitorella) insulin-like growth factor-I.

A full-length cDNA encoding insulin-like growth factor-I (IGF-I) was cloned from mud carp (Cirrhinus molitorella) liver tissue using reverse transcription polymerase-chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. The IGF-I precursor cDNA consists of 822 bp in size with a 218 bp 5'-untranslated region and 118 bp 3'-untranslated region. The 486 bp open reading frame encodes a 161 amino acid peptide with a molecular weight of 17.9 kDa. The deduced IGF-I amino acid sequence shared 82.5-97% and 82.5-84% sequence identity with fish and mammalian counterparts, respectively. The mature IGF-I was overexpressed in Escherichia coli, and the expression level of recombinant mcIGF-I reached to 34.1% of the cell total protein. After purification and refolding of recombinant mcIGF-I, growth-promoting effect of recombinant mcIGF-I was investigated, the results showed that the recombinant mcIGF-I significantly enhanced the growth rate of juvenile tilapia. After 6-week treatment, the growth rates of group 1 and 2 were 53 and 67.3% higher than the saline-treated control group. The recombinant mcIGF-I was more effective than recombinant mcGH to enhance the growth rate of juvenile tilapia. The recombinant mcIGF-I-treated fish revealed no significant changes of content of protein, lipid, ash and moisture in muscle.