Differences in the expression profiles of lncRNAs and mRNAs in partially injured anterior cruciate ligament and medial collateral ligament of rabbits

Long noncoding RNAs (lncRNAs), as a novel regulatory factor, are considered to play a vital role in various biological processes and diseases. However, the overall expression profile and biological functions of lncRNAs in the partially injured anterior cruciate ligament (ACL) and medial collateral ligament (MCL) have not been clearly explored. Partially injured models of ACL and MCL were established in 3-month-old healthy male New Zealand white rabbits. Expression of lncRNAs and mRNAs in the ligament tissue was detected by high-throughput sequencing technology, and biological functions of differentially expressed RNAs were evaluated by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Validation of several differentially expressed RNAs was performed using quantitative real-time PCR (qRT-PCR). Protein-protein interaction (PPI) analysis and competitive endogenous RNA (ceRNA) prediction were used to identify interactions among hub genes and the interaction among lncRNAs, miRNAs, and mRNAs. The results showed that compared with the normal group, there were 267 mRNAs and 329 lncRNAs differentially expressed in ACL and 726 mRNAs and 609 lncRNAs in MCL in the injured group. Compared with MCL, 420 mRNAs and 470 lncRNAs were differentially expressed in ACL in the normal group; 162 mRNAs and 205 lncRNAs were differentially expressed in ACL in the injured group. Several important lncRNAs and genes were identified, namely, COL7A1, LIF, FGFR2, EPHA2, CSF1, MMP2, MMP9, SOX5, LOX, MSTRG.1737.1, MSTRG.26038.25, MSTRG.20209.5, MSTRG.22764.1, and MSTRG.18113.1, which are closely related to inflammatory response, tissue damage repair, cell proliferation, differentiation, migration, and apoptosis. Further study of the functions of these genes may help to better understand the specific molecular mechanisms underlying the occurrence of endogenous repair disorders in ACL, which may provide new ideas for further exploration of effective means to promote endogenous repair of ACL injury.

Decreased Serum Maresin 1 Concentration Is Associated With Postmenopausal Osteoporosis: A Cross-Sectional Study

Background: Maresin 1 plays a role in the regulation of inflammation and metabolic diseases in vivo. An increasing number of studies have reported that postmenopausal osteoporosis (PMOP) is associated with inflammation. However, the potential relationship between the serum Maresin 1 content and PMOP is unclear.Aims: 1) To evaluate the Maresin 1 content in postmenopausal women with osteopenia, osteoporosis, or without these conditions (normal group) and 2) to analyze the correlations between Maresin 1 concentrations and bone mineral density (BMD) and bone turnover markers.Methods: In this cross-sectional study, we measured serum Maresin 1 concentrations, serum biochemical parameters, markers of bone metabolism, and BMD of the femoral neck, lumbar spine, and hip in 141 postmenopausal women.Results: We found that serum Maresin 1 in the osteopenia (140.09 ± 30.54 pg/ml) and PMOP (124.68 ± 31.35 pg/ml) groups were significantly lower than those in the normal group (167.38 ± 24.85 pg/ml) (P < 0.05 and P < 0.001). Serum Maresin 1 levels were positively correlated with femoral neck, lumbar spine, and hip BMD (P < 0.001). Meanwhile, Maresin 1 concentrations were positively associated with 25-hydroxyvitamin D [25(OH)D] levels (P < 0.001), but negatively correlated with β-CrossLaps of type 1 collagen containing cross-linked C-telopeptide (β-CTX) (P = 0.002), procollagen type I amino-terminal propeptide (PINP) (P = 0.004), tartrate-resistant acid phosphatase 5b (TRAP-5b) (P = 0.005), and osteocalcin (OC) levels (P = 0.001). Multivariate logistic regression analysis showed that a decrease in Maresin 1 concentration was still associated with osteopenia (P = 0.035) or PMOP (P = 0.016). Maresin 1 levels had a maximum area under curve of 0.820 for osteopenia and 0.746 for PMOP (P < 0.001). Our results showed that the serum Maresin 1 levels were reduced in osteopenia and PMOP patients compared with that in normal subjects, and were the lowest in the PMOP subjects. The results suggest that Maresin 1 may serve as a new non-invasive diagnostic biomarker for PMOP.

Effects and Mechanism of Oxymatrine Combined with Compound Yinchen Granules on the Apoptosis of Hepatocytes through the Akt/FoxO3a/Bim Pathway

The aim of the present study was to investigate the effects and mechanism of oxymatrine (OMT) combined with compound yinchen granules (CYG) on the apoptosis of hepatocytes through the Akt/FoxO3a/Bim pathway in rats with acute liver failure. The rat model of acute liver failure was established using lipopolysaccharide/D-galactosamine (LPS/D-GalN). The expression of proteins in rat liver tissues was detected by western blot analysis. The mRNA expression of FoxO3a, Bim, Bax, Bcl-2, and caspase-3 in rat liver tissues was detected by RT-qPCR. The apoptosis rate of rat hepatocytes was determined by flow cytometry. Western blots showed that when compared with the normal group, the expression of p-Akt and p-FoxO3a in the model group was decreased ( P < 0.05 ), while the expression of Bim was increased ( P < 0.01 ). Compared with the model group, the expression of p-Akt and p-FoxO3a in the OMT group and the OMT combined with CYG groups was increased ( P < 0.05 or P < 0.01 ), while the expression of Bim was decreased ( P < 0.05 ). The Bax/Bcl-2 ratio and caspase-3 protein expression in the model group were significantly higher than those in the normal group ( P < 0.01 ). The Bax/Bcl-2 ratio and the expression of caspase-3 protein in the OMT group and the OMT combined with CYG groups were significantly lower than those in the model group ( P < 0.01 ). The results of RT-qPCR were consistent with those of western blot. The results of flow cytometry showed that the apoptosis rate of hepatocytes in the OMT group and the OMT combined with CYG groups was significantly lower than that in the model group ( P < 0.05 or P < 0.01 ). We concluded that LPS/D-GalN can induce apoptosis of hepatocytes in rats with acute liver failure through the Akt/FoxO3a/Bim pathway. OMT combined with CYG inhibits apoptosis of hepatocytes in rats with acute liver failure via the Akt/FoxO3a/Bim pathway.

Study on Toll-Like Receptor 2-Mediated Inflammation-Induced Familial Hypertension Combined with Hyperlipemia and Its Mechanism

According to the latest clinical data, cardiovascular diseases have ranked first in prone diseases, causing 40% of the premature deaths of China’s population. This study aimed to investigate the influence of Toll-like receptor 2- (TLR2-) mediated inflammation on the occurrence and development of familial hypertension combined with hyperlipemia and its related mechanism. Blood specimens from 66 patients undergoing coronary atherosclerosis were collected and grouped, including 22 patients into the control group, 25 into the familial hypertension group, and 19 into familial hypertension combined with hyperlipemia group. In this study, ELISA was conducted for determining the levels of four inflammatory factors of TLR2 and IL-1β, IL-6, TNF-ɑ, and CCL2 in serum and the levels of relevant indicators in mice. C57Bl/6j and genetically engineered C.129(B6)-Tlr2tm1Kir/J mice were given subcutaneous injection of normal saline (wild-saline group), 8-week 40% high-fat diet (wild-high-fat group), and subcutaneous Alzet-implanted angiotensin II micropump supplemented with the research diet (wild-high fat-Ang II group, Tlr2-/--high fat-Ang II group). Blood pressure in mice was recorded consecutively with a noninvasive hemopiezometer for eight weeks. TLR2 and IL-1β, IL-6, TNF-ɑ, and CCL2 in serum of patients with familial hypertension combined with hyperlipemia and the hypertension combined with hyperlipemia mouse model were higher than those in the normal group. Under combined intervention of Ang II and the research diet, mRNA expression related to blood pressure, blood lipid, and fat metabolism in Tlr2-/- genetically engineering mice was significantly lower than that in the wild-high fat-Ang II group. The phosphorylation levels of AKT, IKK, and p65 in mice with hypertension combined with hyperlipidemia were significantly higher than those in normal group. The levels of blood pressure and blood lipid in mice after blocking the AKT or NF-κB pathway were significantly downregulated compared with those in the wild-high fat-Ang II group, with statistically significant differences (both P < 0.05 ). In conclusion, TLR2 regulates inflammation through Akt-NF-κB pathway, thus inducing the occurrence and development of familial hypertension combined with hyperlipemia.

High Mobility Group Box-1 Protein and Interleukin 33 Expression in Allergic Rhinitis

<b><i>Background:</i></b> Allergic rhinitis (AR) is characterized by an inflammatory reaction. High mobility group box 1 (HMGB1) protein and interleukin (IL)-33 are damage-associated molecular pattern molecules and have many characteristics similar to pro-inflammatory cytokines. However, the role of IL-33 and HMGB1 in AR remains unclear. The aim of this study is to explore the role of HMGB1 and IL-33 in AR. <b><i>Methods:</i></b> Twenty patients with AR (AR group) and 10 normal controls (normal group) were enrolled in this study. HMGB1 and IL-33 expression were analyzed by immunohistochemistry in epithelial cells of the inferior turbinate mucosa samples. Then, the human nasal mucosa epithelial cells (HNECs) were cultured in vitro, and the house dust mite allergen (Derp1) was used to stimulate the cells. Quantitative real-time PCR and ELISA assay were performed to detect HMGB1 and IL-33 expression in HNECs. <b><i>Results:</i></b> The expression of HMGB1 and IL-33 in the nasal mucosa was higher in the AR group than in the normal group, with a statistically significant difference (<i>p</i> &#x3c; 0.05). In HNECs of AR, the expression of both HMGB1 and IL-33 in stimulated groups was higher than that in non-stimulated groups. The differences were statistically significant (<i>p</i> &#x3c; 0.05). In addition, they increased gradually with the prolonging time and the concentration of the added Derp1. <b><i>Conclusions:</i></b> The expression of HMGB1 and IL-33 were both increased in AR. HMGB1 and IL-33 may have a close relationship in AR.

Identification of Immune Infiltration, Differentially Expressed Genes, and Signaling Pathways in Fanconi Anemia Based on Bioinformatics Analysis

Background and Objective: Fanconi anemia (FA) patients have a reduced ability to form blood cells, accompanied by multiple congenital malformations, mental retardation, solid tumors, and other symptoms. However, the molecular mechanism that causes FA is unclear, and few studies have addressed the regulatory mechanism of immune infiltration in FA. Here, we aimed to identify differentially expressed genes (DEGs), pathways, and immune infiltration involved in FA using integrated bioinformatics analysis and molecular mechanisms. Methods: The GEO gene chip database was searched for FA low density bone marrow tissue, and the content and proportion of 22 types of immune cells in the FA group and the normal group were analyzed using CIBERSORT. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of FA differentially expressed genes (DEGs) using R language and related package programs was also performed.Results: The expression levels of T cells regulatory (Tregs), M2 macrophages, T cells CD8, dendritic cells resting, and T cells CD4 naïve in FA were higher than in the normal group. Furthermore, the expression levels of naïve B cells, monocytes, and resting mast cells in FA were lower than in the normal group. GO analysis of FA differential genes showed that “neutrophil degranulation,” “neutrophil activation,” and “neutrophil activation involved in immune response,” were most frequently enriched among biological processes, with “specific granule,” “tertiary granule,” “tertiary granule lumen” among cellular components, and “carbohydrate binding” among molecular functions. For the KEGG analysis, “Asthma” was most often enriched.Conclusion: This study obtained useful data related to immune infiltration, DEGs, and gene pathways of FA, and provides new evidence for immunotherapy and clinical assessment of FA patients. These results are potentially a useful reference for subsequent related scientific research.

Distribution Characteristics and Species Diversity of Bacteria in Hepatocellular Carcinoma Tissues

This study was to explore the differences in the distribution and species diversity of bacteria between hepatocellular carcinoma (HCC) tissues and normal liver tissues. 28 HCC patients treated with surgery were selected as the research objects (HCC group), and 19 healthy volunteers with normal physical examinations were included in the control group (Normal group). The tumor specimens were obtained by intraoperative and biopsy puncture, and a 16S ribosomal ribonucleic acid (rRNA) library was constructed. Based on the sequencing data obtained by the IlluminaHi Seq sequencing platform, the differences of bacteria in the liver tissues of the HCC group and the Normal group were analyzed at the level of phyla, family, and genus. The Ace, Chao1, and Shannon of the two groups were compared. The results showed that IlluminaHi Seq sequencing obtained a total of 11,714,659 valid sequences, with an average of 131,625 sequences per sample. The proportions of Bacteroidetes, Firmicutes, and Proteobacteria in HCC group and Normal group were 48.75% versus 34.16%, 37.44% versus 18.02%, and 10.85% versus 39.26%, respectively. The Bacteroidaceae, Prevotellaceae, Lachnospiraceae, and Ruminococcaceae accounted for 22.49%, 20.62%, 16.54%, and 19.93% in Normal group; while those in the HCC tissues accounted for 26.83%, 14.22%, 11.14%, and 13.18%, respectively. The dominant bacteria at the genus level in HCC group and Normal group were Bacteroides and Prevotella-9, with the proportions of 24.19% versus 26.04% and 14.19% versus 8.44%, respectively. The difference in operational taxonomic unit (OTU) numbers of HCC and Normal group were compared and analyzed, which were 1,266 and 1,082, respectively, and the number of common OTU in the two tissues was 875. The Ace in HCC tissue and normal liver tissue were 1063.8±66.79 and 1003.6±52.19, respectively. The Ace in HCC tissue was greater than that in normal liver tissue (P < 0.05). The Chao1 and Shannon in HCC tissue were 1022.9±67.74 and 5.4269±0.3608, respectively; while those in normal liver tissue were 1003.6±66.79 and 5.2842±0.9714, respectively. The Chao1 and Shannon in HCC tissues were much higher than those in Normal group (P < 0.05). It showed that there was no difference in the types of bacterial species in HCC tissues, but the proportions of their flora at the level of phyla, family, and genus changed greatly, which may be related to the occurrence of HCC. This study could provide a reference for the diagnosis and treatment of HCC.

Comparison of Voice-Related Quality of Life for the Elderly with and without Voice Disorders According to Genders by Aging Voice Index-Korean Version

Objectives: The present study was performed to investigate the effect of voice problems on voice related quality of life in the elderly with, without voice disorders, according to genders by using the Aging Voice Index-Korean version (AVI-KR).Methods: The AVI-KR, a translated original version of the Aging Voice Index (AVI) into Korean and verified for reliability and validity, was implemented with 50 elderly people without voice disorders (normal group) and 76 elderly people with voice disorders (patient group). Statistical difference according to the group (normal and patient group) and gender (male and female) were analyzed by using a 2-way ANOVA.Results: The mean total score of the AVI-KR of the normal group was significantly higher than that of the patient group. All of the normal group participants showed under 11.00, the cut-off score of AVI-KR, but 17.1% of the patient group appeared under the cut-off score. The female group showed higher scores than the male group, but the difference was not significant. Also, the gender difference of patient groups did not show a statistical significance.Conclusion: The voice-related quality of life in elderly people showed significant difference according to presence/absence of pathological vocal fold status, but the gender difference due to aging did not cause a difference in voice-related quality of life. Even pathological status of vocal fold did not guarantee a bad influence on voice-related quality of life. Therefore, the assessment of elderly people’s voice problems should be carried out not only with an examiner or expert-centered objective tools; but also patient-centered ones, including self-reporting questionnaires and evaluation tools specialized for the elderly, should be developed continuously.

In Different Gender Groups, What Is the Impact of the Fibular Notch on The Severity of High Ankle Sprain: A Retrospective Study of 240 Cases

Background: The function of the distal tibiofibular ligament on the ankle in the occurrence of high ankle sprain (HAS) has been widely studied. Then, in different genders, the effect of the anatomical morphology of fibular notch (FN) on HAS is unclear. Therefore, on the basis of excluding the anatomical differences caused by gender, we explore the impact of different types of FN on the severity of HAS.Methods: We selected 120 patients and further classified these 120 patients into four HAS groups according to FN depth with deep concave type FN ≥ four mm and shallow flat type FN < four mm. A further 120 normal individuals were served as a control group. FN morphological indicators, tibiofibular distance (TFD), and ankle mortise indexes were measured and compared between patients and control groups.Results: In males with shallow flat type, the Anterior tibiofibular distance (aTFD), Middle tibiofibular distance (mTFD), Posterior tibiofibular distance (pTFD), Front tibial width (FTiW), Middle tibial width (MTiW), Posterior tibial width (PTiW) and Depth of ankle mortise (DOAM) of HAS group were higher than those in normal group (P < 0.05). In males with deep concave type, the aTFD, mTFD and DOAM of patients were significantly higher (P < 0.05). Among females with shallow flat type, the aTFD, mTFD, pTFD, FTiW and MTiW in HAS group were greater than those in normal group (P < 0.05). Among the females with deep concave type, the mTFD and pTFD of patients were higher (P < 0.05).Conclusions: After analyzing the morphological indicators of FN, it is found that in both males and females, HAS patients have significant differences in TFD and certain ankle mortise indexes compared with normal people. But more importantly, the above abnormalities are often more common in HAS patients with shallow flat FN, indicating that shallow flat FN may be related to more serious distal tibiofibular ligament injury and ankle mortise widening, resulting in a worse prognosis.Level of evidence: Level III, retrospective comparative study.

Whole Exome-Sequencing of Pooled Genomic DNA Samples to Detect Quantitative Trait Loci in Esotropia and Exotropia of Strabismus in Japanese

Background: Esotropia and exotropia are two major phenotypes of comitant strabismus. It remains controversial whether esotropia and exotropia would share common genetic backgrounds. In this study, we used a quantitative trait locus (QTL)-sequencing pipeline for diploid plants to screen for susceptibility loci of strabismus in whole exome sequencing of pooled genomic DNAs of individuals. Methods: Pooled genomic DNA (2.5 ng each) of 20 individuals in three groups, Japanese patients with esotropia and exotropia, and normal members in the families, was sequenced twice after exome capture, and the first and second sets of data in each group were combined to increase the read depth. The SNP index, as the ratio of variant genotype reads to all reads, and Δ(SNP index) values, as the difference of SNP index between two groups, were calculated by sliding window analysis with a 4 Mb window size and 10 kb slide size. The rows of 200 “N”s were inserted as a putative 200-b spacer between every adjoining locus to depict Δ(SNP index) plots on each chromosome. SNP positions with depth <20 as well as SNP positions with SNP index of <0.3 were excluded. Results: After the exclusion of SNPs, 12,242 SNPs in esotropia/normal group and 12,108 SNPs in exotropia/normal group remained. The patterns of the Δ(SNP index) plots on each chromosome appeared different between esotropia/normal group and exotropia/normal group. When the consecutive groups of SNPs on each chromosome were set at three patterns: SNPs in each cytogenetic band, 50 consecutive sliding SNPs, and SNPs in 4 Mb window size with 10 kb slide size, p values (Wilcoxon signed rank test) and Q values (false discovery rate) in a few loci as Manhattan plots showed significant differences in comparison between the Δ(SNP index) in the esotropia/normal group and exotropia/normal group. Conclusions: The pooled DNA sequencing and QTL mapping approach for plants could provide overview of genetic background on each chromosome and would suggest different genetic backgrounds for two major phenotypes of comitant strabismus, esotropia and exotropia.