The mechanism by which Naru 3 pill protects against intervertebral disc cartilage endplate degeneration based on network pharmacology and experimental verification.

CONTEXT: Naru 3 pill is a traditional Mongolian medicine for the treatment of intervertebral disc degeneration (IDD), but the mechanism is not yet clear. OBJECTIVE: This study investigated the mechanism of Naru 3 pill in the treatment of IDD. MATERIALS AND METHODS: Active ingredients and related targets of Naru 3 pill, as well as IDD-related genes, were collected from public databases. The analysis was performed by protein‒protein interaction network analysis, gene ontology and Kyoto Gene and Genome Encyclopedia (KEGG) functional enrichment analysis, molecular docking and molecular dynamics simulations. Finally, the network pharmacology results were validated by in vitro experiments. RESULTS: Network analysis showed that sesamin, piperine and ellagic acid were potential key components and CASP3, BAX and BCL2 were key targets. KEGG analysis indicated the apoptotic pathway as a potential pathway. Molecular docking showed that sesamin interacted better with the targets than the other components. The results of molecular dynamics simulations indicated that the three systems BAX-sesamin, BCL2-sesamin and CASP3-sesamin were stable and reasonable during the simulation. In vitro experiments showed that sesamin had the least effect on cell growth and the most pronounced proliferation-promoting effect, and so sesamin was considered the key component. The experiments confirmed that sesamin had antiapoptotic effects and reversed the expression of CASP3, BAX and BCL2 in degeneration models, which was consistent with the network pharmacology results. Furthermore, sesamin alleviated extracellular matrix (ECM) degeneration and promoted cell proliferation in the IDD model. CONCLUSION: The present study suggested that Naru 3 pill might exert its therapeutic and antiapoptotic effects on IDD by delaying ECM degradation and promoting cell proliferation, which provides a new strategy for the treatment of IDD.

Efficacy and Safety of Direct Oral Anticoagulants in Patients with Atrial Fibrillation and Liver Disease: a Meta-Analysis and Systematic Review.

BACKGROUND: Liver disease is associated with increased bleeding risk. The efficacy and safety of direct oral anticoagulants (DOACs) is a subject of contention in atrial fibrillation (AF) patients with liver disease. METHODS: Electronic databases (PubMed, Embase, and Cochrane Library) were searched to retrieve studies on the efficacy and safety of DOACs versus warfarin in AF patients with liver disease from January 1980 to April 2020. A meta-analysis was conducted using a random-effects model. RESULTS: Six studies involving 41,859 patients were included. Compared with warfarin, DOACs demonstrated significant reduction in ischemic stroke (HR, 0.68; 95% CI (0.54-0.86)), major bleeding (0.74 (0.59-0.92)), and intracranial hemorrhage (ICH) (0.48 (0.40-0.58)), with no significant effect on gastrointestinal bleeding (P = 0.893) in AF patients with liver disease. Similar results were observed in regular-dose, reduced-dose, and active liver disease subgroups, albeit Asian patients had a slight reduction in major bleeding (P = 0.055). Furthermore, the pooled estimates of individual DOAC subgroups indicated that dabigatran and apixaban led to greater safety in major bleeding (P < 0.001), ICH (P < 0.001), and gastrointestinal bleeding (P < 0.005) in these patients. The same trends were observed in AF patients with cirrhosis. CONCLUSIONS: Our findings suggest that DOACs significantly reduce the risk of ischemic stroke, major bleeding, and ICH, with no significant effect on the risk of gastrointestinal bleeding in AF patients with liver disease compared with warfarin.

Multiple variants in collagen genes are associated with the susceptibility to lumbar disc herniation in the Chinese population.

PURPOSE: Lumbar disc herniation (LDH) is an important cause of back pain and sciatica, but its aetiology is not fully understood. Single-nucleotide polymorphisms (SNPs) in specific collagen genes are known to increase the risk of lumbar disc degeneration. We performed a case-control study among the Chinese Han population to investigate whether genetic variations in collagen genes were associated with the risk of LDH or not. METHODS: We genotyped SNPs selected from 1000 Genome Projects using Agena MassARRAY technology. Three hundred and eighty-four LDH cases were compared with 384 controls of similar age, using the odds ratio and 95% confidence interval to calculate the susceptibility in several genetic models. RESULTS: Our results revealed that subjects with the rs6122316-C variant of the COL9A3 gene had a higher likelihood of LDH than subjects with the allele T variant in both the codominant and recessive models. In addition, after gender stratification analysis, we found significant associations between rs16970089 and rs740024 and LDH risk in females. Age stratification analysis illustrated that rs16970089 and rs6122316 were also correlated with LDH risk in people over 50 years. The smoking stratification illustrated that rs2071358 and rs740024 had an increased association with LDH risk in smokers. And after drinking stratification, we also observed the significance between rs740024 and LDH risk. CONCLUSIONS: Variants in genes for COL1A1, COL9A3 and COL2A1 significantly influence the risk of LDH. Large and well-designed studies are needed to confirm and explain these conclusions. These slides can be retrieved under Electronic Supplementary Material.

The Impact of CYP24A1 Polymorphisms on Hypertension Susceptibility.

BACKGROUND: Hypertension is one of the leading causes of human death and disability. CYP24A1 regulates vitamin D activity and is closely linked to hypertension. However, the relationship between CYP24A1polymorphisms and hypertension risk remains unclear. METHODS: This case-control study included 503 hypertensive patients and 498 healthy controls from the Chinese Han population. The genotypes of CYP24A1polymorphisms were detected using the Agena MassARRAY method. The association between genetic variations of CYP24A1and hypertension risk was evaluated with odds ratios (OR) and 95% confidence intervals (CI) in genetic models. RESULTS: We found that rs56229249 of CYP24A1significantlydecreased the hypertension risk in homozygote (OR 0.51, 95% CI 0.29-0.91, p = 0.022) and recessive models (OR 0.51, 95% CI 0.29-0.91, p = 0.023). Further stratification analyses indicated that hypertension risk is related to age and sex, rs2762934 polymorphism increases hypertension risk among younger subjects (<61 years), and rs1977297 influences the risk of hypertension among older subjects (≥61 years). In addition, rs2762940 is related to hypertension risk in men, and rs56229249 is a protective factor against hypertension in women. CONCLUSIONS: Our study suggests that genetic variations of the CYP24A1gene were significantly associated with susceptibility to hypertension in the Chinese population.

CHRNA5/CHRNA3 gene cluster is a risk factor for lumbar disc herniation: a case-control study.

BACKGROUND: Lumbar disc herniation, a type of chronic low back pain syndrome, is caused by the lumbar intervertebral disk degeneration. Genetic variation in the CHRNA5/CHRNA3 has shown strong associations with smoking-related diseases. This study's aim is to test whether single-nucleotide polymorphisms in the CHRNA5/CHRNA3 gene are associated with lumbar disc herniation risk. METHODS: The genotype frequency distributions of the polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism in 380 lumbar disc herniation patients (case group) and 400 healthy individuals (control group). Allelic, genotypic, and haplotype analyses were performed. RESULTS: We found that the individuals with rs8040868 CT genotype had a 0.46-fold higher risk of lumbar disc herniation than those with rs8040868 TT genotype, in men group (OR = 0.46, 95% CI 0.25-0.84, p = 0.012). Also among women, rs8040868 CT + CC genotype still reduced the risk of lumbar disc herniation under the dominant model (OR = 0.50, 95% CI 0.28-0.89, p = 0.019). Haplotype analysis showed that compared with the CHRNA5 "TACAACCG" wild-type, the "TACACCCG" haplotype was found to be associated with a decreased risk of lumbar disc herniation (LDH) (OR = 0.79, 95% CI 0.63-1.00, p = 0.047), while, in the less than 50-year-old group, CHRNA5 "TACACCCG" increased the risk of LDH (OR = 1.46, 95% CI 1.01-2.13, p = 0.047). CONCLUSIONS: Our data suggest that gene variance in the CHRNA5/CHRNA3 is associated with risk of lumbar disc herniation in the case-control study.

Association study between matrix metalloproteinase-3 gene (MMP3) polymorphisms and ankylosing spondylitis susceptibility.

BACKGROUND: Ankylosing spondylitis (AS) is the second most common cause of inflammatory arthritis worldwide affecting the axial skeleton. Single nucleotide polymorphisms (SNPs) of matrix metalloproteinase-3 (MMP3) in the development of AS has few been investigated in Chinese population. METHODS: A total of 362 patients with AS and 362 healthy controls were enrolled in the study. Five SNPs in MMP3 genotypes were identified by Agena MassARRAY. Chi-squared tests and genetic model were used to evaluate associations. RESULTS: rs522616 had a significant risk of AS development compared to those with the TT genotype (p = 0.008). By multiple logistic regression models analysis, in codominant model, rs522616 CT genotypes also had a 1.44-fold risk (95% CI = 1.06-1.96, p = 0.008) for AS development compared to those with TT genotypes. In recessive model, the CC genotypes was a significantly reduced AS risk for individuals with TT/CT genotype (OR = 0.64; 95% CI = 0.41-0.99, p = 0.040). CONCLUSION: The present study suggests that MMP3 rs522616 polymorphism is associated with AS susceptibility and MMP3 might be a potential diagnostic biomarker for AS. Further independent studies with larger cohorts are warranted to validate our findings in different populations.

IL1R1 Polymorphisms are Associated with Lumbar Disc Herniation Risk in the Northwestern Chinese Han Population.

BACKGROUND The aim of this study was to assess the association of single-nucleotide polymorphisms (SNPs) in IL1R1 with the risk of lumbar disc herniation (LDH) in the Han population in northwest China. MATERIAL AND METHODS To estimate the association of IL1R1 polymorphisms with LDH risk, Agena MassARRAY was used to determine the genotypes of 498 LDH patients and 463 controls. The association between IL1R1 variants and LDH risk was examined by logistic regression analysis with adjustments for age and gender. Stratification analysis was observed between gender and age with polymorphisms of IL1R1. Haplotype construction and analysis in IL1R1 were also applied to detect the potential association. RESULTS The mutant homozygous genotype in codominant model (AA versus GG, OR=2.37, 95% CI: 1.08-5.21, P=0.001) and in recessive model (AA versus GG/GA, OR=2.82, 95% CI: 1.30-6.12, P=0.005) of rs956730 were associated with an increased LDH risk in males, while rs956730 heterozygous genotype under codominant model (AG versus GG, OR=0.65, 95% CI: 0.46-0.92, P=0.001) was a protective genotype in males. In addition, the recessive model (CT/CC versus TT, OR=3.43, 95% CI: 1.11-10.57, P=0.020) of rs10490571 was associated with an increased LDH risk among people older than 50 years of age. CONCLUSIONS This study demonstrated that genetic variants in the IL1R1 genes were associated with LDH risk in the Han population of northwestern China.

MiR-210 inhibits hypoxia-induced apoptosis of smooth muscle cells via targeting MEF2C.

BACKGROUND: Vascular smooth muscle cell (VSMC) apoptosis plays an important role in vascular remodeling and atherosclerotic plaque instability. Growing evidence suggests that microRNAs (miRNAs) play a critical role in VSMC function, however, the underlying mechanism remains unclear. METHODS: This study used a hypoxic-induced VSMC apoptosis model. Expression of miR-210, its target MEF2C, and other key factors of apoptosis were detected and measured by real-time PCR and western blot. Luciferase reporter assay was performed to detect the miR-210 target. The function of miR-210 in apoptosis was determined using flow cytometric cell apoptosis assays. The relationship between miR-210 and MEF2C was confirmed and key apoptosis factors were detected. RESULTS: The restoration of miR-210 function in cells transfected with a miR-210 mimic inhibited VSMC apoptosis compared to control. MiR-210 overexpression inhibited the expression of Bax, Bad, cleaved Caspase-3, and promoted the expression of Bcl-2, Caspase-3, Caspase-9 and mitochondrial cytochrome c at both the mRNA and protein levels. Results also found that MEF2C was a direct target of miR-210 in hypoxic VSMCs. Further, miR-210 suppressed MEF2C expression by directly binding to its 3'-untranslated region and the expression of miR-210 was negatively correlated with MEF2C mRNA levels. CONCLUSIONS: Results from this study provide the first evidence that miR-210 can inhibit apoptosis by targeting MEF2C in hypoxic VSMCs and may support the development of new biomarkers and therapeutic targets for atherosclerosis.

Genetic variants in COL11A2 of lumbar disk degeneration among Chinese Han population.

BACKGROUND: Lumbar disk disease (LDD) is a common musculoskeletal disorder. Several predisposing genetic and environmental risk factors have been established for symptomatic LDD. METHODS: We conducted a case-control association study to investigate the role of the COL11A2 gene in LDD. Genotyping of 384 Chinese Han LDD patients and 384 Chinese Han controls was made for six single-nucleotide polymorphisms (SNPs) from COL11A2 by Agena Massarray. We evaluated these SNPs association with LDD using the chi-square test and genetic model analysis. RESULTS: The strongest associations with LDD were observed for polymorphisms in rs2071025. Carriers of "A" allele had an increased risk of LDD (OR = 1.47, 95% CI = 1.20-1.80, p = 0.0002) as compared with the "G" allele in allele model. We found that rs2071025 were associated with LDD in female and male from the stratification analyses (p < 0.05). Genetic models showed that rs986522(C) significantly increased the risk of LDD in female; however, in males, we did not find significant associations between the rs986522 and LDD risk. CONCLUSION: This study showed a genetic association with COL11A2 polymorphism in individuals with LDD. These data may provide novel insights into the pathogenesis of LDD, although further studies with larger numbers of participants worldwide are needed for validation of our conclusions.

Associations between Variants in BDNF/BDNFOS Gene and Lumbar Disc Herniation Risk among Han Chinese People.

Lumbar disc herniation (LDH) is a low back pain disorder and associated with several single nucleotide polymorphisms (SNPs). However, the role of brain-derived neurotrophic factor (BDNF) and BDNFOS gene in LDH susceptibility remains unknown. To examine whether the variants contribute to LDH, 7 SNPs were genotyped in 380 patients and 692 healthy controls among Han Chinese population. Multiple genetic models, stratification by age/gender and haploview analysis was used by calculating odds ratio (OR) and 95% confidence intervals (CIs). Rs11030064 in BDNFOS gene was associated with modified susceptibility for LDH at age ≤50 years but three loci (rs6265, rs11030104 and rs10767664) of BDNF gene increased LDH risk at age >50 years. Further, rs11030096 polymorphism in BDNFOS gene was associated with LDH the increased susceptibility of LDH in females. Haplotype analysis shown that haplotype "GCC" in the block (rs988712, rs7481311, and rs11030064) increased LDH risk (OR = 1.49, 95% CI = 1.06-2.10, p = 0.022) at age ≤50 years. However, there was no significant association between BDNF/BDNFOS gene and LDH risk in the overall before stratified analysis. For the first time, our results provide evidence on polymorphism of BDNF / BDNFOS gene associated with LDH risk in Chinese Han population.

Association between IL4, IL6 gene polymorphism and lumbar disc degeneration in Chinese population.

Lumbar disc disease (LDD) is a common musculoskeletal disorder, caused by degeneration of intervertebral discs of the lumbar spine and is one of the most common musculoskeletal disorders affliction in adult. There is growing evidence that LDD has strong genetic determinants. We analyze whether the IL4 and IL6 gene polymorphism is related to LDD in Chinese Han population. The participants were 498 with LDD and 463 without LDD. IL4 and IL6 gene polymorphism were determined by Sequenom MassARRAY. We found that SNPs rs1800796(OR = 1.29, 95% CI, 1.07 - 1.57, p = 0.009), rs1524107(OR = 1.28, 95% CI, 1.05 - 1.55, p = 0.013), rs2069840 (OR = 1.39, 95% CI, 1.03 - 1.89, p = 0.033) in IL6 gene were significantly associated with LDD risk at a 5% level. In addition, genetic models found IL4 gene (rs2243250) were associated with LDD. In this study, we analyzed and associated SNPs of IL4 and IL6 with LDD risk. In summary, four variations (rs1800796, rs1524107, rs2069840, rs2243250) of the selected candidate SNPs were associated with susceptibility to LDD in our study. The results of this study have the guiding significance in clinical work in the future in the treatment of lumbar disc herniation patients, not one-sided that the symptoms of low back pain only from mechanical oppression.

Identification of novel biomarker and therapeutic target candidates for acute intracerebral hemorrhage by quantitative plasma proteomics.

BACKGROUND: The systematic mechanisms of acute intracerebral hemorrhage are still unknown and unverified, although many recent researches have indicated the secondary insults. This study was aimed to disclose the pathological mechanism and identify novel biomarker and therapeutic target candidates by plasma proteome. METHODS: Patients with AICH (n = 8) who demographically matched healthy controls (n = 4) were prospectively enrolled, and their plasma samples were obtained. The TMT-LC-MS/MS-based proteomics approach was used to quantify the differential proteome across plasma samples, and the results were analyzed by Ingenuity Pathway Analysis to explore canonical pathways and the relationship involved in the uploaded data. RESULTS: Compared with healthy controls, there were 31 differentially expressed proteins in the ICH group (P < 0.05), of which 21 proteins increased while 10 proteins decreased in abundance. These proteins are involved in 21 canonical pathways. One network with high confidence level was selected by the function network analysis, in which 23 proteins, P38MAPK and NFκB signaling pathways participated. Upstream regulator analysis found two regulators, IL6 and TNF, with an activation z-score. Seven biomarker candidates: APCS, FGB, LBP, MGMT, IGFBP2, LYZ, and APOA4 were found. Six candidate proteins were selected to assess the validity of the results by subsequent Western blotting analysis. CONCLUSION: Our analysis provided several intriguing pathways involved in ICH, like LXR/RXR activation, acute phase response signaling, and production of NO and ROS in macrophages pathways. The three upstream regulators: IL-6, TNF, LPS, and seven biomarker candidates: APCS, APOA4, FGB, IGFBP2, LBP, LYZ, and MGMT were uncovered. LPS, APOA4, IGFBP2, LBP, LYZ, and MGMT are novel potential biomarkers in ICH development. The identified proteins and pathways provide new perspectives to the potential pathological mechanism and therapeutic targets underlying ICH.