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The U6 promoter is an important element driving sgRNA transcription in the CRISPR/Cas9 system. Seven PqU6 promo-ter sequences were cloned from the gDNA of Panax quinquefolium, and the transcriptional activation ability of the seven promoters was studied. In this study, seven PqU6 promoter sequences with a length of about 1 300 bp were cloned from the adventitious roots of P. quinquefolium cultivated for 5 weeks. Bioinformatics tools were used to analyze the sequence characteristics of PqU6 promoters, and the fusion expression vectors of GUS gene driven by PqU6-P were constructed. Tobacco leaves were transformed by Agrobacterium tumefaciens-mediated method for activity detection. The seven PqU6 promoters were truncated from the 5'-end to reach 283, 287, 279, 289, 295, 289, and 283 bp, respectively. The vectors for detection of promoter activity were constructed with GUS as a reported gene and used to transform P. quinquefolium callus and tobacco leaves. The results showed that seven PqU6 promoter sequences(PqU6-1P to PqU6-7P) were cloned from the gDNA of P. quinquefolium, with the length ranged from 1 246 bp to 1 308 bp. Sequence comparison results showed that the seven PqU6 promoter sequences and the AtU6-P promoter all had USE and TATA boxes, which are essential elements affecting the transcriptional activity of the U6 promoter. The results of GUS staining and enzyme activity test showed that all the seven PqU6 promoters had transcriptional activity. The PqU6-7P with a length of 1 269 bp had the highest transcriptional activity, 1.31 times that of the positive control P-35S. When the seven PqU6 promoters were truncated from the 5'-end(PqU6-1PA to PqU6-7PA), their transcriptional activities were different in tobacco leaves and P. quinquefolium callus. The transcriptional activity of PqU6-7PA promoter(283 bp) was 1.59 times that of AtU6-P promoter(292 bp) when the recipient material was P. quinquefolium callus. The findings provide more ideal endogenous U6 promoters for CRISPR/Cas9 technology in ginseng and other medicinal plants.
Subject(s)
Panax/genetics , Promoter Regions, Genetic , Agrobacterium tumefaciens/genetics , Computational Biology , Cloning, MolecularABSTRACT
@#[Objective] To investigate the evolution of inflammation under conditions and the effects of ginsenosides on macrophages subjected to the simulated weightlessness, with the aim of mitigating the inflammation. [Methods] Initially, genes related to weightlessness, inflammation, and immunity were identified in the GeneCards database. Then, Search Tool for the Retrieval of Interaction Gene/Proteins (STRING) protein network analysis was conducted to determine the core targets involved in the weightlessness-induced inflammation. Subsequently, Label-Free Quantitative (LFQ) proteomics was carried out to discern the distinctive genes within ginsenoside-treated Tohoku Hospital Pediatrics-1 (THP-1) cells. Next, utilizing the outcomes of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, the biological processes and signaling pathways in which ginsenosides predominately engaged were scrutinized, and the primary targets of ginsenosides in combating weightlessness-induced inflammation were examined. Finally, enzyme-linked immunosorbent assay (ELISA) was performed to detect the secretion levels of interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α from lipopolysaccharide (LPS)-induced THP-1 cells under simulated weightlessness conditions, as well as during the weightlessness recovery period following treatment with ginsenosides. [Results] A total of 2 933 genes associated with inflammation, 425 genes linked to weightlessness, and 4 564 genes connected to immunity were retrieved from the GeneCards database. Protein-protein interaction (PPI) networks were generated to identify pivotal targets associated with weightlessness-induced inflammation such as IL-1β, IL-6, TNF, and albumin (ALB). It was found that ginsenosides primarily participated in the regulation of various inflammationrelated signaling pathways and pathways related to pathogenic microorganism infections. Moreover, it has a significant impact on the expression of proteins such as cluster of differentiation 40 (CD40), IL-1β, and poly ADP-ribose polymerase 1 (PARP1). As revealed in the simulated weightlessness cell test, ginsenosides exhibited a remarkable capacity to attenuate the secretion of inflammatory factors, specifically IL-6 and TNF-α (P < 0.000 1), in THP-1 macrophages following induction by LPS under simulated weightlessness conditions. In addition, it reduced the secretion of IL-1β, IL-6, IL-8, and TNF-α (P < 0.000 1) during the weightlessness recovery phase [Conclusion] Weightlessness can disrupt several inflammation-related signaling pathways, but ginsenosides were shown to mitigate the release of various inflammatory factors in macrophages subjected to simulated weightlessness, thereby exerting a protective role against inflammation. This study has laid a theoretical groundwork for further exploring the potential application of ginsenosides in safeguarding against LPS induced inflammation in a weightlessness environment.
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Objective@#To explore the therapeutic effects of ginseng total saponins (GTSs) on cognitive impairments in astronauts caused by prolonged exposure to microgravity environment.@*Methods@#Fifty specific pathogen-free (SPF) male Wistar rats were randomized into control, hindlimb suspension (HLS), Huperzine A (HLS-Hup A 0.1 mg/kg), low-dose GTSs (HLS-GTSs 100 mg/kg), and high-dose GTSs (HLS-GTSs 200 mg/kg) groups, based on the completion time of reward-directed conditioning tasks. Except for rats in the control group, the others were subjected to HLS and treated with drugs (day 20 – 58), received reflex test under the condition of rewarding, and underwent Nissl body staining and Western blot detection on hippocampal.@*Results@#After modeling, rats in HLS group exhibited a reduction in the number of lever presses and an increase in the completion time of the reward-directed operant conditioning task Ⅰ (P < 0.05) when compared with the control group, which were not substantially altered in the HLS-GTSs 100 and 200 mg/kg groups (P > 0.05). In the reward-directed operant conditioning task Ⅱ, the HLS group rats demonstrated a marked decrease in the number of lever presses (P < 0.05) and nose pokes (P < 0.01) when compared with the control group rats; the HLS-GTSs 100 mg/kg showed a significant increase in the number of lever presses and nose pokes (P < 0.05), while the HLS-GTSs 200 mg/kg demonstrated a significant reduction in completion time and an elevation in the number of lever presses (P < 0.05) when compared with the HLS group rats. In visual signal discrimination task, compared with the control group rats, the HLS group rats showed decrease in the indexes of the visual signal discrimination(P < 0.01), while HLS-GTSs 100 and 200 mg/kg groups exhibited manifest increase in it (P < 0.01). In reward extinction experiment, the number of lever presses in HLS rats significantly increased when compared with the control group (P < 0.01); compared with the HLS group, HLS-GTSs 100 and 200 mg/kg groups demonstrated a marked descrease (P < 0.05). The expressions of N-methyl-D-aspartic acid receptor 1 (NR1) and phosophorylated N-methyl-Daspartic acid receptor 2B (p-NR2B) proteins were markedly decreased in rats in the HLS group (P < 0.05 and P < 0.01, respectively), while that of NR2B protein maintained the same (P > 0.05). GTSs increased the expression levels of p-NR2B (P < 0.01).@*Conclusion@#GTSs improved the learning and memory ability of complex operations by regulating the NR1/NR2B phosphorylation pathways in rats.
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@#High myopia complicated with fundus lesions is one of irreversible blinding eye diseases. Posterior seleral staphyloma(PSS)is one of the most basic pathologies in a series of complications of high myopia. This article reviews the pathogenesis, examination methods, classification and treatment of PSS in high myopia by sorting out domestic and foreign literature, providing a better understanding of the prevention and control of PSS.
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@#Objective To study the therapeutic effects of Shenyuan Gan (参远苷, SYG) on the inflammatory response in BV2 microglial cells induced by lipopolysaccharide (LPS). Methods The cytotoxicity of SYG to BV2 microglial cells was evaluated using a Cell Counting Kit-8 (CCK-8) assay, and the effect of SYG concentrations on LPS-induced BV2 microglial cells was studied. The morphological changes were observed using an optical microscope. The nitric oxide (NO) concentration in cell culture supernatant was determined using Griess reagent. The expression of cytokines and inflammatory mediators were also measured by an enzyme-linked immunosorbent assay (ELISA). Western blot analysis was used to determine the levels of inducible NO synthase (iNOS), nuclear factor-kappa B (NF-κB) p65, alpha inhibitor of NF-κB (IκB-α), phosphorylation-IκB-α (p-IκB-α), NOD-like receptor 3 (NLRP3), and caspase-1 expression. Moreover, the expression of iNOS, NLRP3, and ionized calcium binding adapter molecule 1 (Iba1) was also observed using immunofluorescent staining. Results SYG had a low cytotoxic effect on BV2 microglial cells and could significantly decr-ease LPS-induced morphological changes of BV2 microglial cells (P < 0.05). ELISA results showed that SYG significantly inhibited the LPS-induced increase in interleukin (IL)-1β and IL-6 in BV2 microglia cells (P < 0.05), and Western blot analysis showed that the phosphorylation levels of iNOS, NF-κB p65, and IκB-α as well as NLRP3 and caspase-1 expression were also significantly decreased, and IκB-α expression was increased after SYG treatment (P < 0.05, compared with the LPS-treated group). The immunofluorescence results were consistent with the Western blot results, and Iba1 staining indicated that the cell morphology tended to be resting. These results indicate that SYG has a certain inhibitory effect on LPS-induced inflammation in BV2 microglial cells. Conclusion SYG can inhibit LPS-induced release of inflammatory factors in BV2 microglial cells by affecting the phosphorylation levels of NF-κB p65 and IκB-α. SYG is a valuable candidate for treating neuroinflammation-related diseases.
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Abstract Inflammation-related immune responses and bone metabolism lead to extensive tooth loss in periodontitis. Objective: This study aims to investigate the effect of peroxisome proliferator-activated receptor (PPAR) alpha agonist anti-inflammatory treatment in vitro and in ligature-induced experimental periodontitis in vivo . Methodology: Splenocytes were isolated from C57BL/6J mice and cultured for 48 hours under the following conditions: control, P. gingivalis lipopolysaccharide (LPS) (1 µg/ml); experimental, LPS (1 µg/ml) + PPARα agonist (fenofibrate) at 1, 10, 50, 100 µM. MRNA and secreted protein levels of TNF-α expression were detected by RT-qPCR and ELISA, respectively. Silk ligatures (7-0) were tied around maxillary second molars of C57BL/6J mice for two weeks. Optimized doses of fenofibrate (50 µM) and vehicle control were injected into the contralateral side of the palatal gingiva on days three, six, and nine. At day 14, bone resorption, osteoclastogenesis, and gingival mRNA expression levels of TNF-α, IL-1β, IL-6, and RANKL/OPG were measured by micro-computed tomography, Tartrate-resistant acid phosphatase (TRAP) staining, and Real-time quantitative PCR, respectively. Results: TNF-α expression in cultured spleen cells were significantly increased in the presence of LPS, when compared with the control group, and significantly reduced by fenofibrate treatment in a dose-dependent manner from 1-100 µM (p<0.05). Gingival mRNA levels of TNF-α, IL-1β, IL-6, and the ratio of RANKL/OPG, were significantly decreased after injection of fenofibrate, when compared to the control side (p<0.05). Periodontal bone loss and TRAP positive cell formation were significantly decreased on the side with an injection of fenofibrate, as compared to the control side (p<0.05). Conclusions: An anti-inflammatory treatment, PPARα agonist, inhibited inflammation and periodontal bone loss in ligature-induced experimental periodontitis.
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Abstract Objective To explore the molecular mechanism of neuropathologic damage induced by radiofrequency ablation at different temperatures. Methods This is basic research, and 36 SD rats were used to construct the neuropathological injury model. The rats were subjected to radiofrequency stimulation at different temperatures and were divided into 6 groups according to the temperature injury: 42°, 47°, 52°, 57°, 62°, and 67°C groups. Conduction time, conduction distance, and nerve conduction velocity were recorded after temperature injury. HE-staining was used to observe the histopathological morphology of the sciatic nerve. The expression of SCN9A, SCN3B, and NFASC protein in sciatic nerve tissue were detected by western blot. Results With the increase in temperature, nerve conduction velocity gradually decreased, and neurons were damaged when the temperature was 67°C. HE-staining showed that the degrees of degeneration of neurons in rats at 47°, 52°, 57°, 62°, and 67°C were gradually increased. The expression of SCN9A, SCN3B protein in 57°, 62°, 67°C groups were much higher than that of NC, 42°, 47°, 52°C groups. However, the expression of NFASC protein in 57°, 62°, 67°C groups was much lower than that of the NC, 42°, 47°, 52°C groups. Conclusion There was a positive correlation between temperature caused by the radiofrequency stimulation to neuropathological damage. The mechanism is closely related to the expression of SCN9A, SCN3B, and NFASC protein in nerve tissue caused by heat transfer injury.
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Abstract Objective The purpose of this study is to investigate the pathogenic role of PPARα in periodontal antigen treated gingival cells in vitro and in experimental periodontitis in vivo . Methodology Gingival fibroblasts, gingival epithelial cells and splenocytes were isolated from C57BL/6J wild type (WT) mice and treated with fixed P. gingivalis at for 48 hours. The mRNA levels of PPARs, TNFα, IL-1β and IL-10 were detected by Real-time quantitative PCR. Silk ligatures after being soaked in the P.gingivalis suspension were tied around both maxillary second molars of WT mice or PPARα knock-out (KO) mice for two weeks. PPARα agonist fenofibrate and vehicle control were injected into the different side of the palatal gingiva on days 3, 6, and 9. At day 14, bone resorption and gingival mRNA expression levels of PPARs, TNFα, IL-1β and IL-10 were measured by micro-computed tomography and RT-qPCR respectively. Results P. gingivalis treatment downregulated the expression of PPARα, but not PPARβ or PPARγ, and increased the expression of TNF-α and IL-1β in Gingival fibroblasts, gingival epithelial cells and splenocytes from WT mice. Gingival mRNA levels of PPARα were significantly decreased in experimental periodontitis in WT mice. The bone loss of PPARα KO mice in experimental periodontitis was significantly higher than WT mice and was not reduced by fenofibrate treatment. Gingival TNFα protein expressions were significantly increased by P. gingivalis associated ligation and decreased by fenofibrate treatment in WT mice but not in PPARα KO mice. Conclusion This study suggests that PPARα plays an essential role in periodontitis.
Subject(s)
Humans , Alzheimer Disease , Cohort Studies , Disease Progression , Neuropsychological TestsABSTRACT
Reportedly, circular RNAs (circRNAs) are crucial regulators in cancer progression. Nonetheless, the molecular mechanism of circRNAs in hepatocellular carcinoma (HCC) has not been fully clarified. Gene expression omnibus (GEO) database was employed to screen out the differentially expressed circRNAs in HCC. qRT-PCR and western blot were executed to detect circ_0001806 expression, miR-193a-5p expression, and MMP16 mRNA and protein expressions in HCC. The effect of circ_0001806 on HCC was analyzed by the CCK-8 method and Transwell experiment. RIP assay, pull-down experiment, and dual-luciferase reporter gene experiment were applied to validate the targeting relationships among circ_0001806, miR-193a-5p, and MMP16. Circ_0001806 was up-modulated in HCC tissues and cell lines. Knockdown of circ_0001806 impeded the multiplication, migration, and invasion of HCC cells. Circ_0001806 could up-regulate MMP16 expression through repressing miR-193a-5p, thereby facilitating the malignant biological behaviors of HCC. Circ_0001806 promoted HCC progression by regulating miR-193a-5p/MMP16 axis.
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Modeling a protein functional network in concerned species is an efficient approach for identifying novel genesin certain biological pathways. Tea plant (Camellia sinensis) is an important commercial crop abundant innumerous characteristic secondary metabolites (e.g., polyphenols, alkaloids, alkaloids) that confer tea qualityand health benefits. Decoding novel genes responsible for tea characteristic components is an important basisfor applied genetic improvement and metabolic engineering. Herein, a high-quality protein functional networkfor tea plant (TeaPoN) was predicted using cross-species protein functional associations transferring andintegration combined with a stringent biological network criterion control. TeaPoN contained 31,273 nonredundant functional interactions among 6,634 tea proteins (or genes), with general network topologicalproperties such as scale-free and small-world. We revealed the modular organization of genes related to themajor three tea characteristic components (theanine, caffeine, catechin) in TeaPoN, which served as strongevidence for the utility of TeaPoN in novel gene mining. Importantly, several case studies regarding geneidentification for tea characteristic components were presented. To aid in the use of TeaPoN, a concise webinterface for data deposit and novel gene screening was developed (http://teapon.wchoda.com). We believe thatTeaPoN will serve as a useful platform for functional genomics studies associated with characteristic secondarymetabolites in tea plant.
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OBJECTIVE@#To compare the clinical therapeutic effect of long-snake moxibustion and ginger-partitioned moxibustion at point on nonspecific low back pain (NLBP) with symptom of cold and dampness.@*METHODS@#A total of 120 patients were randomized into a long-snake moxibustion group, an ashi point group and a waiting for treatment group, 40 cases in each one. Ginger-partitioned moxibustion was applied from Dazhui (GV 14) to Yaoshu (GV 2) of governor vessel in the long-snake moxibustion group, and was applied at point of affected area in the ashi point group, 40 min each time, once every other day and totally 8 times were required. No intervention was adopted in the waiting for treatment group, and after the trial, long-snake moxibustion was applied. Before and after treatment, the visual analogue scale (VAS) scores of rest and activity, the Oswestry disability index (ODI) score and the score of cold and dampness symptom were observed in the 3 groups.@*RESULTS@#Compared before treatment, the VAS scores of rest and activity, the ODI scores and the scores of cold and dampness symptom after treatment were decreased in the long-snake moxibustion group and the ashi point group (<0.05). After treatment, the variations of the above indexes in the long-snake moxibustion group and the ashi point group were larger than those in the waiting for treatment group (<0.05), and the variations of the above indexes in the long-snake moxibustion group were larger than those in the ashi point group (<0.05).@*CONCLUSION@#Long-snake moxibustion can effectively improve the pain, dysfunction and the symptom of cold and dampness in patients with nonspecific low back pain, and the improvement is superior to the ginger-partitioned moxibustion at point.
Subject(s)
Humans , Acupuncture Points , Low Back Pain , Therapeutics , Medicine, Chinese Traditional , MoxibustionABSTRACT
Objective@# To explore the influence of marital status on the survival of patients with buccal mucosa squamous cell carcinoma (BMSCC) and to provide a scientific basis for risk assessment and nursing decisions for these patients.@*Methods @# A total of 1 006 BMSCC participants were identified in the Surveillance, Epidemiology, and End Results(SEER) database. Kaplan-Meier analysis was used to evaluate the effect of marital status (married, unmarried and unhappily married) on the survival of patients with buccal squamous cell carcinoma. The independent prognostic effect of marital status was further determined by univariate and multivariate Cox proportional hazard regression analysis.@*Results@# Marital status was an independent prognostic factor for BMSCC patients (P=0.001). Married patients had the best survival, while separated/divorced/widowed patients had the worst survival (HR=1. 445, P=0.001), and never married patients had an intermediate risk (HR=1.297, P=0.040).@*Conclusion@#Marital status was an independent prognostic factor for BMSCC patients. Unmarried and unhappily married patients had worse survival, which suggests that we should pay more attention to patients’ marital status, especially unmarried and unhappily married patients.
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Acute lymphocytic leukemia (ALL) is one of the most common malignancies, especially in young people. Combination chemotherapy for ALL typically includes corticosteroids (Kantarjian et al., 2000). Hyperglycemia is a well-recognized complication of corticosteroids, and chemotherapy-induced diabetes (CID) is not uncommon (27.5%-37.0%) during the treatment of ALL (Hsu et al., 2002; Weiser et al., 2004; Alves et al., 2007). Besides the effect of corticosteroids, potential factors triggering hyperglycemia in ALL also include direct infiltration of the pancreas by leukemia cells and β cell dysfunction induced by chemotherapeutic agents such as L-asparagine (Mohn et al., 2004).
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Age Factors , Antineoplastic Agents/adverse effects , Diabetes Mellitus/chemically induced , Precursor Cell Lymphoblastic Leukemia-Lymphoma/mortalityABSTRACT
Abstract The fruits of Xanthium strumarium L., Asteraceae, have been used for various diseases in Chinese folk medicine, including allergic rhinitis, tympanitis, arthritis, ozena etc. The current study was aimed to investigate the therapeutic effects of caffeoylquinic acids from fruits of X. strumarium on allergic rhinitis in animals. The toxicity test indicated that the caffeoylquinic acids have no obvious toxicity. By using HPLC assays combined with reference standards, ten caffeoylquinic acids were identified as the predominant constituents. Anti-allergic activities of the caffeoylquinic acids were evaluated using passive cutaneous anaphylaxis test and Schultz-Dale test; dimethylbenzene induced ear edema test was performed to evaluate its anti-inflammatory effect. Then, the allergic rhinitis model in rats was established to evaluate the therapeutic effects of the caffeoylquinic acids against allergic rhinitis with the following indexes: allergic rhinitis symptom scores, serum levels of pro-inflammatory cytokines, histopathological examination, and histamine release. Our study revealed that the caffeoylquinic acids showed obvious anti-allergic and anti-inflammatory properties, and its treatments were beneficial for ameliorating the nasal symptoms, decreasing pro-inflammatory cytokines, and inhibiting the releases of histamine. Collectively, the caffeoylquinic acids might be utilized as effective and safe disease therapeutic agents for allergic rhinitis.
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Objective@#Based on the dual-factor model of mental health, this study aims to explore the effects of acceptance and commitment therapy on the mental health of high school students.@*Methods@#During Oct. to Nov. 2017, General Health Questionnaire12(GHQ-12) and Depression Anxiety and Stress Scale21(DASS-21) was administered among 543 high school students from a middle school in Nanchang. According to the inclusion and exclusion criteria, 20 high school students were screened and were randomly assigned to the intervention group and control group, respectively. The intervention group received one intervention (1.5 h) per week for 8 weeks. The control group did not receive the intervention. Pre-and post-intervention assessment were conducted using the Acceptance and Action Questionnaire-2nd Edition (AAQ-II), the Cognitive Fusion Questionnaire-Fusion (CFQ-F), the Satisfaction With Life Scale (SWLS), the Positive Affect Scale (PAS), the GHQ-12, and the DASS-21.@*Results@#After intervention, the post-test scores of AAQ-II, CFQ-F, GHQ-12, and DASS-21 in the intervention group significantly decreased (t=2.91, 2.47, 4.91, 4.42, P<0.05) and the post-test score on PAS significantly increased (t=-5.27, P<0.01). There were no statistical changes in AAQ-II, CFQ-F, GHQ-12, and DASS-21 scores in the control group after intervention (t=-1.25, -0.09, -0.61, -0.69, -0.81, P>0.05).@*Conclusion@#Acceptance and commitment therapy can effectively improve the psychological flexibility and positive emotions, as well as alleviate general psychological distress and negative emotions, and promote the improvement of mental health of high school students.
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Cystic echinococcosis (CE) in sheep is a hazardous zoonotic parasitic disease that is caused by Echinococcus granulosus (Eg). At present, serological test is an important diagnostic method for Eg infection in domestic animals. Here, a fusion protein Eg mefAg-1 harboring 8 dominant B-cell epitopes of Eg such as antigen B, tetraspanin 1, tetraspanin 6, reticulon and Eg95 was produced in E. coli and evaluated for CE in sheep by indirect ELISA. Eg mefAg-1 showed in ELISA a high sensitivity (93.41%) and specificity (99.31%), with a coincidence rate of 97.02%. Overall, it is suggested that the Eg mefAg-1 could be a potential antigen candidate for CE serodiagnosis in sheep.
Subject(s)
Animals, Domestic , Echinococcosis , Echinococcus granulosus , Enzyme-Linked Immunosorbent Assay , Epitopes, B-Lymphocyte , Methods , Parasitic Diseases , Sensitivity and Specificity , Serologic Tests , SheepABSTRACT
Objective: To assess the level of serum lipid metabolism related hormones in the patients of obstructive sleep apnoea hypoxia syndrome (OSAHS) combine with non-alcoholic fatty liver disease (NAFLD), and explore the potential role in the pathogenesis. Methods: Patients that main complaint for snoring were selected. According to polysomnography and NAFLD screening, the patients were divided as follows: OSAHS+NAFLD group, OSAHS group, NAFLD group, and control group, and the levels of serum lipid metabolism related hormones were measured. The patients with OSAHS were divided into three groups, i.e., mild, moderate and severe groups according to the degree of apnea hypopnea index (AHI) and minimal oxyhemoglobin saturation (MinSaO2) respectively. The statistical analysis about the morbidity of NAFLD in the patients with different OSAHS severity were analyzed. Multiple regression analysis was used to determine the independent predictors of NAFLD. Results: Significant differences were found in AHI, MinSaO2, the duration of hemoglobin desaturation (TSaO2<90%), body mass index, adiponectin, leptin, high sensitive C reactive protein and lipopolysaccharide (P<0.05 for all) among the groups of OSAHS, NAFLD and OSAHS+NAFLD. The morbidity of NAFLD exited statistical difference in the patients with different OSAHS severity (17.0%, 51.6% and 97.2%, P=0.006), and the same trend was seen in the patients with different MinSaO2 severity (18.2%, 57.1% and 91.9%, P=0.011). Multiple regression analysis showed that body mass index, AHI and adiponectin were independent predictors of the morbidity of NAFLD (P=0.006, P=0.020 and P=0.008). Conclusion: The morbidity of NAFLD increases with the worsening of OSAHS. There is positive correlation between the severity and the level of serum adiponectin and leptin in the patients with OSAHS and NAFLD.
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<p><b>OBJECTIVE</b>To investigate the effect of Bryan cervical disc replacement on cervical curvature and replacement segment in the treatment of cervical spondylosis, and analyze its clinical efficacy.</p><p><b>METHODS</b>Forty-nine patients underwent Bryan artificial cervical disc replacement from August 2010 to August 2013 were retrospectively analyzed. There were 26 males and 23 females, aged from 27 to 66 years old with an average of 48.5 years. Including 21 cases of nerve root type, 16 cases of spinal cord type, 12 cases of mixed type. And 21 cases were single segment replacement, 28 cases were double segments replacement. Preoperative and postoperative pain was compared using visual analogue scale(VAS);cervical function was observed by Japanese Orthopedic Association Scores (JOA) and Neck Disability Index (NDI);cervical lordosis curvature, function spinal unit(FSU) curvature, activities of replacement and adjacent segment, FSU activity were measured by cervical X-rays. Clinical effects were evaluated by the Odom method.</p><p><b>RESULTS</b>All the patients were followed up for 18.5 to 37.3 months with an average of 33.1 months. Pvoperative VAS, JOA, NDI scores were 7.08±1.55, 5.2±1.9, 39.96±7.06 before operation, 3.76±2.33, 13.20±1.20, 25.20±6.64 at 3 months after operation, and 2.80±1.50, 14.3±1.6, 24.24±7.89 at the last follow-up. Patients at three months and the last follow-up after operation were obviously improved in their VAS, JOA, and NDI scores (<0.05). There was no significant difference between the scores at last follow-up and 3 months after operation. Cervical lordosis changed from (10.64±4.26)° preoperatively to (13.68±4.56) ° at the last follow-up;the FSU curvature from (5.40±0.41) °to (9.92±2.00) °at the last follow-up (<0.05). The range of motion of the cervical spine preoperatively, 3 month postoperatively, and at the last follow-up were(70.84±6.17)°, (60.00±6.58)°, (71.48±4.61)°; FSU activities were(12.00±0.49)°, (9.36±0.26)°, (12.52±0.33)°;the activities of replacement segment were (10.48±0.67)°, (7.24±0.34)°, (9.28±0.36)°;the activities of upper segment of replacement were (10.52±0.60)°, (8.60±0.30)°, (10.44±0.43)°;the activities of lower segment of replacement were (8.48±0.40)°, (6.56±0.36)°, (9.60±0.39)°;there were significant differences in above items preoperatively and 3 months postoperatively(<0.05); and there were no significant difference preoperatively and at the last follow-up(>0.05). Pharyngeal discomfort and hoarseness occurred in 5 cases, and were recovered within 2 weeks after operation;heterotopic ossification occurred in 1 case at 6 months after operation;displacement of prosthesis occurred in 1 case at 12 months after operation. Based on Odom standard to evaluate the clinical outcome, 20 cases obtained excellent results, with 27 good, 2 general.</p><p><b>CONCLUSIONS</b>Bryan artificial cervical disc replacement not only generate good clinical effects, but may also restore the cervical lordosis and FSU curvature, while retaining the activities of replacement segment, and restoring overall cervical biomechanical functions.</p>
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A simple, sensitive and reliable method was developed for simultaneous determination of ten banned drugs residues including zeranols(ZALs),chloroamphenicol,pentachlorophenol,etc. in swine urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The urine samples were pretreated using lyophilization and QuEChERS procedures, respectively. Acetonitrile and ammonium acetate (5 mmol/L) were chosen as mobile phases. Target compounds were separated well in ZorbaxSB-C18by following the optimized gradient elution program and determined by LC-MS/MS in negative electrospray ionization mode. The linearity of the matrix-matched standard curve of ten analytes in two methods was good in the range of the experimental concentration with correlation coefficients more than 0.99. The recoveries of ten drugs were in the range of 80.7%-107.7% and 73.5%-103.3% at the spiked levels of 5,10 and 20 μg/L by lyophilization and QuEChERS methods,respectively. The coefficients of variation were less than 15%. The limits of detection (LOD) and the limits of quantification (LOQ) from lyophilization and QuEChERS method were 0.1 to 2.0 μg/L and 0.2 to 5.0 μg/L,respectively.