Construction of artificial neural network (ANN) based on predictive value of prognostic nutritional index (PNI) and neutrophil-to-lymphocyte ratio (NLR) in patients with cervical squamous cell carcinoma.

To explore the analytical worth of prognostic nutritional index (PNI) and neutrophil-to-lymphocyte ratio (NLR) in patients with cervical squamous cell carcinoma. The clinical data of 539 patients with cervical cancer in the Affiliated Tumor Hospital of Nantong University from December 2007 to October 2016 were analyzed retrospectively. The ROC is used to select the best cutoff values of PNI and NLR, which are 48.95 and 2.4046. Cox regression analysis was used for univariate and multivariate analysis. Survival differences were assessed by Kaplan-Meier (KM) survival method. Finally, a 3-layer artificial neural network (ANN) model is established. In cervical squamous cell carcinoma, the KM survival curve showed that the overall survival (OS) rate of high-level PNI group was significantly higher than that of low-level PNI group (P < .001), while the OS rate of low-level NLR group was significantly higher than that of high-level NLR group (P = .002). In non-squamous cell carcinoma, there was no significant difference in OS between the 2 groups (P > .005). According to Cox multivariate analysis, preliminary diagnosed PNI and NLR were independent prognostic factors of cervical squamous cell carcinoma (P < .001, P = .008), and pathological type and International Federation of Gynecology and Obstetrics (FIGO) stage also had a certain impact on tumor progression (P = .042, P = .048). The increase of PNI and the decrease of NLR will help patients with cervical squamous cell carcinoma live longer. ANN showed that PNI and NLR were of great importance in predicting survival. Preoperative PNI and NLR are independent predictors of cervical squamous cell carcinoma patients related to clinicopathological features, and have particular value in judging prognosis.

Proteomics and Phosphoproteomic Analysis to Identify Spleen of Takifugu rubripes Infected Cryptocaryon irritans.

Takifugu rubripes is important commercially fish species in China and it is under serious threat from white spot disease (cyptocaryoniasis), which leads to heavy economic losses. In this study, we used proteomics and phosphoproteomic analysis to identify differentially abundant proteins in the spleen of T. rubripes infected with the Cryptocaryon irritans. We identified 5,307 proteins and 6,644 phosphorylated sites on 2,815 phosphoproteins using high-throughput proteomics analysis of the spleen of T. rubripes based on 26,421 unique peptides and 5,013 modified peptides, respectively. The 5,307 quantified host proteins, 40 were upregulated and 43 were downregulated in the infection group compared to the control group. Among the 2815 phosphoproteins, 44/120 were upregulated/downregulated, and 62/151 were upregulated/downregulated in the 6644 quantified phosphosites. Using the combination of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses, screening for significantly different phosphoproteins, motif analysis and protein-protein interaction analysis, we ultimately identified three phosphorylated proteins (G-protein-signaling modulator 1-like, zinc finger protein 850-like, and histone H1-like) and three phosphorylated protein kinases (serine/threonine-protein kinase homolog isoform X2, mitogen-activated protein kinase 5-like, and protein kinase C theta type) as potential biomarkers for T. rubripes immune responses. We then screened the phosphorylation sites of these biomarker proteins for further verification. Based on our results, we speculate that phosphorylation modification of the phosphorylation sites is involved in the immunity of T. rubripes against C. irritans.

Is Chinese herbal formula (nourishing Yin therapy) effective and well tolerated as an adjunct medication to hydroxychloroquine in the treatment of primary Sjögren's syndrome? A meta-analysis of randomised controlled trials.

BACKGROUND: Previously, data mining methodology was used to identify 71 patented prescriptions in Chinese patent databases, indicating that Yin-nourishing therapy (YNT) may be an adjunct medication to hydroxychloroquine in the treatment of primary Sjögren's syndrome (pSS). The purpose of this study was to investigate effects of the addition of YNT, which includes tonifying liver and kidney therapy (TLKT) and replenishing Qi and nourishing Yin therapy (RQNYT), in the treatment of pSS. METHODS: Fourteen databases (including Chinese, English, Japanese, Korean and Latin databases) were searched to identify randomised controlled trials (RCTs) of YNT plus hydroxychloroquine (YNTPH) versus hydroxychloroquine alone in patients with pSS. Relevant publications up to June 2021 were selected. A meta-analysis and trial sequential analysis (TSA) were performed using Review Manager 5.3, Stata 14.0 and TSA 0.9 software. The quality of identified studies was assessed based on the Cochrane risk of bias tool and GRADE (grading of recommendations, assessment, development and evaluation) criteria. RESULTS: We included five RCTs with a total of 345 participants. Pooled results indicated significant differences in short-term outcomes, which were assessed via European League Against Rheumatism (EULAR) Sjögren's Syndrome Patient Reported Index (ESSPRI), EULAR Sjögren's Syndrome Disease Activity Index (ESSDAI), tear production, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and immunoglobulin G (IgG) results when YNTPH was compared with hydroxychloroquine use alone (p < 0.05). No significant difference in salivary flow rate was reported. The most common side effect reported for both groups was gastrointestinal reaction. Sensitivity analyses suggested that heterogeneity might be ascribed to clinical methodology. Subgroup analyses revealed heterogeneities regarding salivary flow rate were eliminated. TLKT appeared to be better than RQNYT for improving salivary flow rate. TSA only supported changes in ESSPRI, ESSDAI and ESR values. For all studies, the quality of evidence was low. CONCLUSION: YNTPH may be an effective complementary therapy. Current evidence, however, is insufficient for determining whether YNTPH is more effective than hydroxychloroquine alone. Well-designed RCTs are needed to determine the role of YNT in pSS treatment.

Response mechanism of gut microbiome and metabolism of European seabass (Dicentrarchus labrax) to temperature stress.

In animals, the gut microbiome is vital to growth, and changes in the composition of these microbial communities may affect growth and adaptability to the environment. Temperature is another important factor that influences the healthy growth of animals. To date, the mechanism by which juvenile European seabass (Dicentrarchus labrax) and their symbiotic flora adapt to changes in environmental temperature is not well understood. Therefore, we evaluated the effect of temperature on the gut microbiota and metabolism of European seabass juveniles. We used 16S rRNA gene amplicon sequencing and non-targeted liquid chromatography with tandem mass spectrometry (LC-MS/MS)-based metabolomics to study the gut microbes of European seabass after 60 days of rearing of water temperature at 10 °C (T1), 15 °C (T2) and 20 °C (T3). At the phylum level, the abundance of the gut microbiota did not differ significantly among the three groups after 60 days of cultivation. At the genus level, however, the abundance of Faecalibacterium, Filifactor, Butyricicoccus, and Erysipelotrichaceae UCG-006 in the intestines differed significantly among the temperature groups. Compared with T2, the relative abundance of Filifactor in T1 was significantly increased, while Faecalibacterium was significantly decreased, while the relative abundance of Butyricicoccus and Erysipelotrichaceae UCG-006 in T3 was significantly increased. The LC-MS/MS analysis revealed 107 metabolites in the 10 °C group and 68 metabolites in the 20 °C group that differed significantly from those in the intestines of fish in the 15 °C control group. These metabolites are closely related to several metabolic pathways, including amino acid metabolism, glucose and lipid metabolism, and the tricarboxylic acid cycle. Correlation analysis of the Intestine microbiota, metabolic pathways, and metabolites identified metabolic pathways and metabolites that were strongly related to the observed significant differences in the microbiome among the temperature groups. These results show that temperature can induce significant changes in the gut microbiota and metabolism of European seabass juveniles, and that significant changes in metabolites may be mediated through the interaction of the microbiome and metabolic pathways.

Impact Resistance Analysis and Optimization of Variant Truss Beam Structure Based on Material Properties.

In order to meet the increasing application requirements with regards to structural impact resistance in industries such as mining, construction, aerospace engineering, and disaster relief and mitigation, this paper designs a variant truss beam structure with a large shrinkage ratio and high impact resistance. Based on the principle of the curved trajectory of scissor mechanisms, this paper conducts a finite element simulation analysis of the impact load on the truss beam structure, a theoretical analysis of the impact response and a relevant prototype bench-top experiment, completing a full study on the impact resistance mechanism of the designed variant truss beam structure under the impact load. In the paper, the buffer effect of the external load impact on the variant truss beam structure is analyzed from the perspective of the energy change of elastic-plastic deformation. This paper proposes an optimization strategy for the variant truss beam structure with the energy absorption rate as the optimization index through extensive analysis of the parameter response surfaces. The strategy integrates analyses on the response characteristic analysis of various configuration materials to obtain an optimal combination of component parameters that ensures that the strength of the truss beam structure meets set requirements. The strategy provides a feasible method with which to verify the effectiveness and impact resistance of a variant truss structure design.

Comparative efficacy and safety of four classical prescriptions for clearing damp-heat recommended by clinical guidelines in treating rheumatoid arthritis: a network meta-analysis.

BACKGROUND: In China, along with conventional synthetic disease-modifying anti-rheumatic drugs (csDMARDs), some herbal formulae for clearing damp-heat are widely applied in treating rheumatoid arthritis (RA). We aimed to summarize and compare the clinical effects of 4 guideline-recommended formulae, including Baihuguizhi decoction, Dangguiniantong decoction, Simiao pill, and Xuanbi decoction. METHODS: PubMed, Cochrane Library, EMBASE, Chinese National Knowledge Infrastructure (CNKI), Chinese Science and Technique Journals (CQVIP), WanFang, and SinoMed (CBM) databases were searched for randomized controlled trials from inception to July 2020 evaluating the efficacy and safety of these recommended herbal formulae combined with 1 csDMARD versus csDMARD alone in RA patients. A pairwise meta-analysis was conducted in RevMan 5.3 software, and a Bayesian network meta-analysis (NMA) was performed with Stata 14.0, R 4.0.2, GeMTC 0.14.3, and JAGS 4.3.0 software. Cochrane Handbook 5.1.0 was used to assess the risk of bias. Publication bias was evaluated using Egger's test, the trim-fill adjustment, and funnel plots. Trial sequential analysis (TSA) was performed to validate the overall results. The rank probability of interventions was calculated and clustered by the surface under the cumulative ranking curve (SUCRA). Pharmacologic actions of formulae were explored through the network pharmacology approach. RESULTS: A total of 15 studies, including 1,079 individuals, were identified. Simiao pill + csDMARD [SMPPD, odds ratio (OR) =6.62, 95% confidence interval (CI): 2.88 to 16.84] was superior to csDMARDs alone in clinical efficiency, and was more able to reduce C-reactive protein and erythrocyte sedimentation rate levels [mean difference (MD) =-7.91, 95% CI: -17.41 to -1.25; MD =-9.31, 95% CI: -14.48 to -5.56 respectively]. Although publication bias was observed (P=0.033), the trim-fill method indicated that the pooled values kept stable. Fewer adverse events (AEs) were shown with SMPPD (6.45%). TSA confirmed the results of efficacy rate at SMPPD. Network pharmacology included 5 common components and 66 common targets among 4 formulae in treating RA, involving regulating immunity and relieving inflammation. DISCUSSION: SMPPD might be a preferable complementary therapy for RA. However, considering the limitations of this study, recommendations for clinical practice should be validated by the results of further well-designed studies.

Development and application of diffusive gradients in thin-films for in situ sampling of the bitterest chemical - denatonium benzoate in waters.

Denatonium benzoate (DB), a commonly used bitter agent in numerous products, has recently been recognized as a waterborne contaminant due to concern about its potential persistence, mobility and toxicity (PMT). However, its occurrence, levels and fate in global aquatic environments are largely unknown. In this study, a new sampling method, based on diffusive gradients in thin films (DGT) with mixed-mode cation exchange (MCX) as the binding agent, was developed for measuring DB in waters. MCX shows a rapid adsorption and high capacity for DB. DB is linearly accumulated by MCX-DGT. pH (6-8), ionic strength (0.01-0.5 M), or DOM (0-10 M) do not show any significant effect on the MCX-DGT performance, confirming its reliability. The DGT measurements in a wastewater treatment plant (WWTP) are comparable to those by paralleled grab sampling. The field results suggest DB is persistent in WWTPs and could be a potential domestic wastewater indicator. Therefore, MCX-DGT is a promising technique for understanding the environmental occurrence, levels and fate of DB. This is a first report of using DGT for DB monitoring and of DB occurrence in Chinese environments. Further exploration of DGT as a reliable passive monitoring tool for a wide range of PMT substances in different applications is warranted.

Administration of a recombinant ALDH7A1 (rA7) indicates potential regulation of the metabolite and immunology pathways in Atlantic salmon infected with Aeromonas salmonicida.

The bacterium Aeromonas salmonicida is the pathogen responsible for furunculosis, which is a serious disease of salmonids. This disease has a significant economic impact on the economic benefits of the global salmon farming industry. However, the pathogenesis of this disease in fish is still unknown. Members of the aldehyde dehydrogenase gene (ALDH) superfamily play a key role in the enzyme detoxification of endogenous and exogenous aldehydes. In this study, we obtained a recombinant aldehyde dehydrogenase 7A1 (ALDH7A1) protein to find its functions on Atlantic salmon infected by A. salmonicida. The transcriptional response in the liver of Atlantic salmon (Salmo salar) with differing levels of A. salmonicida infection was analysed and compared in order to reveal mechanisms by which ALDH7A1 may confer infection resistance. With the addition of ALDH7A1 protein, it was found that a total of 13,369 genes were annotated with one or more KEGG and localized to 360 KEGG pathways in the high concentration infection group. The differential expression genes were more enriched in immune signalling pathways such as the Toll-like receptor signalling pathway, NF-kappa B signalling pathway and TNF signalling pathway. On the other hand, at low concentrations of infection, KEGG enriched a smaller number of differential expression genes. However, these differential genes were more concentrated in immune signalling pathways such as the PI3K-Akt signalling pathway, JAK-STAT signalling pathway and complement and coagulation cascades. In addition, several known immune-related genes including HSP90α, HSP70, DNA damage-inducible transcript 4, integrin alpha 5 and microtubule-associated protein 2 were among the differentially expressed transcripts. These data provide the first insights into the host-ALDH7A1 vaccine interactome. The results of this study contribute to identifying the potential resistance mechanisms of Atlantic salmon to A. salmonicida infection and determining future treatment strategies.

HSP70 gene expression responses to the temperature stress in pufferfish (Takifugu rubripes).

In this study, we isolated and characterized HSP70 cDNA from pufferfish (Takifugu rubripes). The 3053 bp full-length TrHSP70 sequence consisted of a 167 bp 5'-UTR (untranslated region), a 2535 bp open reading frame, and a 351 bp 3'-UTR. BLAST analysis revealed that the TrHSP70 shared high similarity with HSP70 sequences in other species. In our study, we set 3 experimental groups as H1 group (20 °C), H2 group (24 °C), and H3 group (28 °C) for checking the expression level of TrHSP70 in T. rubripes. Tissue-specific gene expression results showed that TrHSP70 had higher expression in the intestines than other tissues of the T. rubripes by RT-qPCR. In the experimental group, we found that the expression of TrHSP70 was upregulated in different tissues in the H3 group. The results show that TrHSP70 is a constitutively expressed gene, which plays an important role in maintaining normal physiological function and coping with stress.

Adoption of a new gait system to evaluate the clinical effects of minimally invasive needle-knife scope therapy for the treatment of rheumatoid arthritis of the knee joint.

BACKGROUND: The purpose of this study was to use kinematic gait analysis, to evaluate the clinically curative effects of minimally invasive needle-knife scope therapy in the treatment of rheumatoid arthritis (RA) of the knee joint. METHODS: A total of 32 patients with RA of the knee joint were treated with minimally invasive needle-knife scope therapy. All patients were assessed with a kinematic gait analysis system, and then analyzed and compared the measurements with the kinematic gait performances of 28 healthy middle-aged and elderly participants. RESULTS: Before receiving the therapy, the ranges of motion (ROMs) of the patients' tibial inward and outward rotation, flexion and extension, anterior and posterior displacement of the tibia, and tibial upward and downward movement were all smaller than those of the healthy middle-aged and elderly group (P<0.05). After receiving the therapy, the patients' knees had a remarkably larger ROM. After one month of this therapy, they had a noticeably greater ROM in flexion and extension, tibial inward and outward rotation, and upward and downward movement of the tibia (P<0.05). After 1 month of therapy, the patients' maximum flexion angle and tibial posterior displacement angle were noticeably greater than beforehand (P<0.01). After 1 year of therapy, the patients' flexion and extension range became remarkably greater than it was after 1 month of the therapy (P<0.01). Their ranges of DOF reflected no apparent difference to the healthy middle-aged and elderly group, which remained smaller than the latter (P<0.05). After 1 year of therapy, the maximum values of the patients' flexion angle, tibial internal rotation angle, tibial posterior displacement angle, and tibial downward movement angle were remarkably higher than before commencing treatment (P<0.05). CONCLUSIONS: Minimally invasive needle-knife scope therapy enables a good recovery of function for patients with RA and remarkably improves the DOF of knees. Gait analyses are more objective, accurate, and quantitative than other indexes, and thus may become a new means to assess the changes in knee joint functions.

Nondestructive Damage Testing of Beam Structure Based on Vibration Response Signal Analysis.

Nondestructive damage-testing technology based on vibration signal analysis makes full use of the response characteristics of wave and energy. With the advantages of wide bandwidths of response frequency and high sensitivity, the nondestructive testing technology based on vibration signal analysis has a superiority in the application for the detection and characterization of structural defects, and has become one of the important methods for the nondestructive testing of structural material defects and damage. This paper presents a novel method of detection localization and quantitative analysis for local damage in beam structures, based on the response analysis of vibration signals. A damage-detection and -identification algorithm based on a unscented Kalman filter (UKF) was designed, which greatly reduces the computational workload in the process of damage identification over that in conventional methods. The method presented in this paper has significances to widen the application scope of the nondestructive testing method, and increase the recognition efficiency and effectiveness of this kind of method in engineering.

Quantitative proteomic analysis in serum of Takifugu rubripes infected with Cryptocaryon irritans.

Cryptocaryon irritans can cause cryptocaryonosis (white spot disease) in marine fish but the pathogenesis of the disease is unclear. In this work, we used high-throughput proteomics to identify differentially expressed proteins in the serum of Takifugu rubripes challenged with C. irritans. By using quantitative proteomic assays combined with Tandem Mass Tag-labeled quantitative proteomic analysis, we identified a total of 2088 differentially abundant proteins (1706 proteins were quantified, p < 0.05, fold-change threshold ≥ 2), including 21 up-regulated and 44 down-regulated. Combined with STRING-based functional analysis, we ultimately obtained eight proteins including glucokinase-like, integrin beta-1-like isoform X2, H4, H2A.V, histone H1-like, histone H2AX-like, histone H2B 1/2-like and myosin-9 isoform X1, which could be considered as potential biomarkers for T. rubripes immune responses. Eight proteins that were selected to validate significant differentially expressed genes at the proteomic level were consistent with qPCR at the transcriptomic level. The proteins identified in our work may serve as candidates for elucidating the molecular mechanism of cryptocaryonosis in T. rubripes. Our collective findings could provide new insights into searching for disease-specific targets and biomarkers, which may be effective indicators of C. irritans infection in T. rubripes.

Characterization and expression analysis of the C8α and C9 terminal complement components from pufferfish (Takifugu rubripes).

C8α and C9 mediate the membrane attack complex formation and bacterial lysis and are important components in the complement system. The cDNA sequences of the C8α and C9 genes were cloned from Takifugu rubripes. The full-length cDNA of Tr-C8α was 1893 bp and included a 5'-UTR of 69 bp and 3'-UTR of 83 bp. The full-length cDNA of Tr-C9 was 2083 bp and included a 5'-UTR of 72 bp and 3'-UTR of 250 bp. The expression of Tr-C8α and Tr-C9 was detected in newly fertilized eggs of T. rubripes. The expression of these two genes was at a higher level in the liver than in other tissues tested. After lipopolysaccharide (LPS) challenge, the gene expression of Tr-C8α and Tr-C9 increased more significantly in the liver. With these combined results, we further understood how Tr-C8α and Tr-C9 function in the innate immunity of pufferfish. Our findings could deepen the understanding of immune regulation in pufferfish.

Gentiopicroside exerts convincing antitumor effects in human ovarian carcinoma cells (SKOV3) by inducing cell cycle arrest, mitochondrial mediated apoptosis and inhibition of cell migration.

PURPOSE: Gentiopicroside is an important plant secondary metabolite and has been reported to exhibit tremendous pharmacological potential. In the present study we investigated the anticancer effects of gentiopicroside against ovarian SKOV3 cancer cells. METHODS: Anticancer effects were determined by MTT assay. Apoptosis was investigated by DAPI and annexin V/propidium iodide (PI) double staining. Mitochondrial membrane potential (MMP) determination and cell cycle analysis were carried out by flow cytometry. Protein expression was examined by western blotting. RESULTS: The results showed that gentiopicroside exerted anticancer effects on SKOV3 cancer cells in a dose-dependent manner. The IC50 of gentiopicroside was 20 µM against the SKOV3 cancer cells. The anticancer effects were mainly found to be due to loss of MMP and induction of apoptosis. The Bax/Bcl-2 expression ratio was also altered upon gentiopicroside treatment. Furthermore, gentiopicroside could arrest the SKOV3 cells in G2/M phase of the cell cycle and prevent their migration and invasion. CONCLUSIONS: These results indicate that gentiopicroside could prove a potential lead molecule in the treatment and management of ovarian cancer.

PVA-encapsulated Palladium Nanoparticles: Eco-friendly and Highly Selective Catalyst for Hydrogenation of Nitrobenzene in Aqueous Medium.

In aqueous medium without any other additives, palladium (Pd) nanoparticles with water-soluble polyvinyl alcohol (PVA) as stabilizer were synthesized for the catalytic hydrogenation of nitrobenzene. Under the optimum experimental conditions, the nitrobenzene conversion and the selectivity for aniline were 99.3 % and 100 %, respectively. Comprehensive characterization methods, including TEM, UV/Vis, confocal laser scanning microscopy (CLSM), XRD and XPS allowed a better understanding of the role of PVA aggregates and the properties of Pd nanoparticles. The nitrobenzene conversion exceeded 80 % even after 6 cycles without any treatment of the catalyst. A mechanism about the hydrogenation of nitrobenzene catalyzed by Pd/PVA system was proposed. The Pd/PVA catalyst also exhibited excellent activity and selectivity, particularly to ortho-fluoronitrobenzene and ortho-nitrotoluene. This research can provide a reference for the environmentally friendly catalysis for hydrogenation of nitrobenzene and other substituted nitrobenzene compounds.

[Efficacy of fan-needle radiofrequency for knee osteoarthritis guided by meridians-muscles theory].

OBJECTIVE: To observe the efficacy difference between fan-needle radiofrequency guided by meridians-muscles theory and oral administration of medication for knee osteoarthritis (KOA). METHODS: One hundred patients with KOA were randomly divided into an acupuncture group and a medication group, 50 cases in each one. The patients in the acupuncture group were treated with fan-needle radiofrequency guided by meridians-muscles theory at tendon nodes around knee joints (ashi points), 4 points per treatment, once every two weeks and two treatments were given. The patients in the medication group were treated with oral administration of celecoxib capsules, 1 capsule every day for 4 weeks. The visual analogue scale (VAS) and Western Ontario and McMaster Universities (WOMAC) in the two groups were observed before treatment, after treatment and during 4-week follow-up visit. The clinical efficacy was evaluated. RESULTS: Compared before treatment, the VAS was reduced in the two groups after treatment and during 4-week follow-up visit (all P<0.05), and the VAS score in the acupuncture group was lower than that in the medication group (both P<0.05). Compared before treatment, the pain score, stiffness score, activity function score and total score of WOMAC were reduced in the two groups after treatment and during 4-week follow-up visit (all P<0.05), and the scores in the acupuncture group were lower than those in the medication group (all P<0.05). After treatment, the total effective rate was 80.0% (40/50) in the acupuncture group, which was superior to 56.0% (28/50) in the medication group (P<0.05). During 4-week follow-up visit, the total effective rate was 76.0% (38/50) in the acupuncture group, which was superior to 40.0% (20/50) in the medication group (P<0.05). CONCLUSION: The efficacy of fan-needle radiofrequency guided by meridians-muscles theory is superior to oral administration of celecoxib capsules for KOA, which could relieve joint pain and stiffness, improve joint mobility, and has long effective duration.

Lysophosphatidic Acid Promotes Expression and Activation of Matrix Metalloproteinase 9 (MMP9) in THP-1 Cells via Toll-Like Receptor 4/Nuclear Factor-κB (TLR4/NF-κB) Signaling Pathway.

BACKGROUND Lysophosphatidic acid (LPA) is an active compound of oxidized low-density lipoprotein that serves as an endogenous TLR4 ligand. Ligand activation of TLR4 activates nuclear factor-kappaB (NF-κB) and the transcription of NF-κB-regulated inflammatory cytokines, which are involved in the development of atherosclerosis. MMP9 is a member of the MMP family and can affect plaque stability. However, the mechanism responsible for the effect of LPA on the expression and activation of MMP9 has not been fully elucidated. In the present study we examined the effect of LPA on MMP9 expression and activity in THP-1 cells and the involvement of Toll-like receptor 4/nuclear factor-κB (TLR4/NF-κB) signaling pathway in this effect. MATERIAL AND METHODS Human THP-1 cells were treated with 0-10 µM LPA for 4 h, or treated with 1 µM LPA for 0-8 h, and were then transfected with TLR4-specific siRNA or treated with 20 µg/ml cafestol acid phenethyl ester (CAPE, an NF-κB inhibitor). MMP9 mRNA and protein levels were measured by quantitative RT-PCR and Western blot analysis, respectively, and MMP9 activity was measured by zymography. RESULTS LPA upregulated MMP9 mRNA and protein levels and MMP9 activity in THP-1 cells in both concentration- and time-dependent manners. Transfection of cells with TLR4-siRNA-2 or treatment with CAPE significantly inhibited the upregulated MMP9 expression and activation. This inhibition was further enhanced by combining the TLR4-siRNA-2 transfection and CAPE pretreatment. CONCLUSIONS LPA can promote MMP9 expression and enhance MMP9 activity in THP-1 cells, in part via the TLR4/NF-kB signaling pathway.

The effect of lysophosphatidic acid on Toll-like receptor 4 expression and the nuclear factor-κB signaling pathway in THP-1 cells.

Toll-like receptors (TLRs) are major receptors that mediate the innate immune and inflammatory responses, of which TLR4 has been found most closely related to human atherosclerosis. After ligands are polymerized and activated by TLR, the mitogen-activated protein kinase and nuclear factor-κB (NF-κB) pathways are activated, leading to promotion of NF-κB-regulated transcription of inflammatory factors, thus playing a role in the physiological and pathological processes in atherosclerosis. Oxidized lipoproteins or their components, oxidized lipids, have been confirmed as endogenous TLR receptors. Lysophosphatidic acid (LPA) is an active component of low-density lipoprotein that induces vascular endothelial lesions. However, the mechanism of the TLR4/NF-κB signaling system involved in LPA-induced atherosclerosis has not been fully elucidated. In this study, we investigated the effects of LPA on TLR4 expression, nuclear translocation of NF-κB p65 subunit, and changes in the cytokine tumor necrosis factor α (TNF-α) in human THP-1 cells. LPA upregulated expression of the TLR4 mRNA and protein in THP-1 cells in a dose- and time-dependent manner, induced NF-κB p65 activation synchronously in THP-1 cells, and increased TNF-α secretion. After TLR4 was blocked using TLR4 monoclonal antibody, NF-κB p65 expression and TNF-α secretion were inhibited significantly. These data suggest that LPA can significantly upregulate TLR4 expression and promote NF-κB activation and proinflammatory cytokine secretion in THP-1 cells; it is possible that the TLR4/NF-κB signaling pathway mediates the atherogenic effect of LPA.

[Treatment of Knee Osteoarthritis by Tendons of Minimally Invasive Therapy Combined Drug Ther- apy: a Clinical Observation of Sixty Cases].

OBJECTIVE: To assess the efficacy of tendons of minimally invasive therapy (TMIT) combined drug therapy by comparing it with treatment by drug therapy alone on patients with knee osteoarthritis (KOA). METHODS: Totally 60 KOA patients were assigned to the treatment group and the control group according to random digit table, 30 in each group. Patients in the control group took Hydrochloric Acid Glucosamine Capsule and Celecoxib Capsule. Patients in the treatment group additionally received TMIT. The treatment course for all was 4 weeks. Scores for visual analogue scale (VAS) and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index were observed and recorded at week 1 and 4 after treatment by acupotomology mirror. RESULTS: Compared with before treatment, improvement was shown in VAS score, pain and stiffness degrees, activities and functions, and WOMAC scores at week 1 and 4 after treatment in all patients with statistical difference (P < 0.05). Besides, better effect was shown in the treatment group (P < 0.05). CONCLUSIONS: TMIT combined drug therapy could relieve KOA patients' pain, stiffness and joint activities, elevate the overall efficacy. TMIT was easily operated with less injury.

Relationship between lysophosphatidic acid and matrix metalloproteinase-9 plasma concentrations and carotid atheromatous plaque stability in patients with cerebral infarction.

OBJECTIVE: To investigate the correlation between plasma lysophosphatidic acid (LPA), matrix metalloproteinase (MMP)-9 and carotid atheromatous plaque stability in patients with cerebral infarction. METHOD: Patients with cerebral infarction underwent carotid artery duplex ultrasonography and transcranial Doppler monitoring for detection of microemboli. Patients were stratified by plaque type (no plaque, intima thickening, unstable plaque, stable plaque) and presence or absence of micoremboli. Plasma LPA and MMP-9 were quantified. RESULT: LPA and MMP-9 concentrations were significantly higher and microemboli positivity was significantly more common in patients with unstable plaque (n = 21) than the other three groups (intima thickening, n = 16; stable plaque n = 41; no plaque; n = 12). There was a significant positive correlation between LPA and MMP-9 concentrations. CONCLUSION: Plasma LPA and MMP-9 concentrations may be useful biomarkers in the clinical identification and prediction of unstable plaque, and in guiding treatment.