Circular RNA hsa_circ_0000175 Serves as a Potential Biomarker for Rheumatoid Arthritis via miR-31-5p/GSDME Axis.

Rheumatoid arthritis (RA) is a common inflammatory autoimmune disease characterized by synovial inflammation and joint damage. Previous studies have shown that pyroptosis plays an important role in the pathogenesis of RA. In this study, the effects of circular RNA hsa_circ0000175 on pyroptosis and inflammation of RA were evaluated. Serum levels of circ_0000175 and miR-31-5p were determined by RT-qPCR, and the correlation between them was evaluated by Spearman correlation analysis. Fibroblast-like synoviocytes (FLSs) were extracted and prepared for in vitro study. The subcellular localization of circ_0000175 was detected by FISH assay. Pyroptosis and inflammatory cytokines interleukin (IL)-1ß, IL-18 and IL-6 were measured by flow cytometry and ELISA, respectively. RNA pull-down and luciferase reporter assays verified the interaction between circ_0000175 and miR-31-5p. Western blot was used to detect the differential expression of pyroptosis-related factors (GSDME-N, GSDMD-N, cleaved caspase-1 and cleaved caspase-3). Circ_0000175 level was increased but miR-31-5p expression was decreased in PBMCs of RA patients and LPS/ATP-treated FLSs, companied with negative correlation. Moreover, miR-31-5p was a target of circ_0000175 in RA-FLSs. Silencing of circ_0000175 or overexpression of miR-31-5p significantly alleviated LPS/ATP-induced pyroptosis in FLSs through both caspase-1/GSDMD and caspase-3/GSDME pathways. Additionally, GSDME was identified as the target of miR-31-5p. The inhibitory effects of circ_0000175 depletion on pyroptosis and inflammation in RA-FLSs treated with LPS/ATP were strengthened by GSDME knockdown. Circ_0000175 can induce pyroptosis and trigger inflammatory response during the occurrence of RA through the miR-31-5p/GSDME axis, which provides a novel therapeutic target for RA treatment.

Diosmetin induces apoptosis and protective autophagy in human gastric cancer HGC-27 cells via the PI3K/Akt/FoxO1 and MAPK/JNK pathways.

Gastric cancer represents a significant global health concern, necessitating the exploration of novel therapeutic options. Diosmetin, a natural flavonoid derived from citrus and vegetables, has demonstrated promising anti-tumor activity against various tumor cells. However, the potential anticancer effect of diosmetin in gastric cancer and its underlying mechanism have yet to be elucidated. In this study, we aimed to investigate the impact of diosmetin on cell proliferation, migration, cell cycle progression and apoptosis in human gastric cancer HGC-27 cells. Our findings revealed that diosmetin effectively suppressed cell proliferation, induced G2/M phase cell cycle arrest, and triggered cell apoptosis. Mechanistically, diosmetin downregulated the expression of antiapoptotic proteins Bcl-2 and Bcl-xL, while upregulated the level of proapoptotic proteins such as Bax, cleaved PARP and cleaved caspase-3. Additionally, diosmetin inhibited Akt and FoxO1 phosphorylation, while activated the MAPK signaling pathway. Notably, pretreatment of IGF-1, an Akt activator, attenuated the diosmetin-induced apoptosis. Furthermore, pretreatment with SP600125, a JNK inhibitor, significantly reduced the protein level of LC3B, while promoted the expression of cleaved caspase-3 and cleaved PARP. Collectively, our results suggest that diosmetin holds promise as an effective therapeutic agent against gastric cancer by inducing apoptosis through inhibition of the Akt/FoxO1 pathway and promoting protective autophagy via the MAPK/JNK signaling pathway.

The effect of perioperative mmunonutrition on patients undergoing esophagectomy: a systematic review and updated meta-analysis / Efectos de la inmunonutrición perioperatoria en pacientes sometidos a esofagectomía: revisión sistemática y metaanálisis actualizado

Background: immunonutrition has been introduced and proposed to have positive modulating effects on inflammatory and immune responses in surgical patients. This meta-analysis aimed to assess whether perioperative enteral immunonutrition (EIN) can reduce postoperative complications or reduce inflammatory responses in esophageal cancer (EC) patients undergoing esophagectomy. Methods: PubMed, Embase, Web of science, EBSCO, and Cochrane library databases were systematically searched. Randomized controlled trials (RCTs) assessing the effect of EIN before and/or after surgery in EC patients undergoing esophagectomy were identified. Two investigators independently searched articles, extracted data, and assessed the quality of included studies. Results: ten RCTs involving 1,052 patients were included in the meta-analysis, including 573 patients in the EIN group and 479 patients in the enteral nutrition (EN) group. Overall, no significant difference was observed between the two groups in the incidence of postoperative pneumonia, surgical site infection, intra-abdominal abscess, septicemia, and urinary tract infection. No significant incidence of postoperative anastomotic leakage, acute respiratory distress syndrome (ARDS), and in-hospital mortality was found. Conclusions: perioperative enteral immunonutrition did not reduce the incidence of infectious complications and anastomotic leakage in EC patients undergoing esophagectomy, nor did it reduce postoperative CRP and IL-6, but did not increase in-hospital mortality.(AU)

Antecedentes: se ha introducido y propuesto la inmunonutrición para regular activamente la inflamación y la respuesta inmune en pacientesquirúrgicos. El presente metaanálisis fue diseñado para evaluar si la inmunonutrición enteral perioperatoria (EIN, por sus siglas en inglés) puedereducir las complicaciones postoperatorias o la inflamación en pacientes con cáncer de esófago (CE) sometidos a esofagectomía.Métodos: se realizó una búsqueda sistemática en las bases de datos de PubMed, Embase, Web of Science, EBSCO y Cochrane Library. Se evaluóel efecto de la EIN preoperatoria y/o postoperatoria en un ensayo aleatorizado controlado (RCT) en pacientes con cáncer de esófago sometidosa esofagectomía. Dos investigadores buscaron independientemente artículos, extrajeron datos y evaluaron la calidad de los artículos incluidos.Resultados: el metanálisis incluyó diez ensayos controlados aleatorios en los que participaron 1.052 pacientes, de los cuales 573 fueronincluidos en el grupo EIN y 479, en el grupo de nutrición enteral (NE). En general, no hubo diferencia significativa en la incidencia de neumoníapostoperatoria, infección del sitio quirúrgico, absceso intraperitoneal, sepsis e infección del tracto urinario entre los dos grupos. No hubo diferenciasignificativa en la incidencia de fístula anastomótica postoperatoria, síndrome de distrés respiratorio agudo (SDRA) y mortalidad hospitalaria.Conclusión: la inmunonutrición enteral perioperatoria no puede reducir la incidencia de complicaciones infecciosas postoperatorias y fístulasanastomóticas, ni la PCR postoperatoria ni la IL-6. Pero no aumentó la mortalidad hospitalaria.(AU)

The effect of perioperative immunonutrition on patients undergoing esophagectomy: a systematic review and updated meta-analysis.

Introduction: Background: immunonutrition has been introduced and proposed to have positive modulating effects on inflammatory and immune responses in surgical patients. This meta-analysis aimed to assess whether perioperative enteral immunonutrition (EIN) can reduce postoperative complications or reduce inflammatory responses in esophageal cancer (EC) patients undergoing esophagectomy. Methods: PubMed, Embase, Web of science, EBSCO, and Cochrane library databases were systematically searched. Randomized controlled trials (RCTs) assessing the effect of EIN before and/or after surgery in EC patients undergoing esophagectomy were identified. Two investigators independently searched articles, extracted data, and assessed the quality of included studies. Results: ten RCTs involving 1,052 patients were included in the meta-analysis, including 573 patients in the EIN group and 479 patients in the enteral nutrition (EN) group. Overall, no significant difference was observed between the two groups in the incidence of postoperative pneumonia, surgical site infection, intra-abdominal abscess, septicemia, and urinary tract infection. No significant incidence of postoperative anastomotic leakage, acute respiratory distress syndrome (ARDS), and in-hospital mortality was found. Conclusions: perioperative enteral immunonutrition did not reduce the incidence of infectious complications and anastomotic leakage in EC patients undergoing esophagectomy, nor did it reduce postoperative CRP and IL-6, but did not increase in-hospital mortality.

Introducción: Antecedentes: se ha introducido y propuesto la inmunonutrición para regular activamente la inflamación y la respuesta inmune en pacientes quirúrgicos. El presente metaanálisis fue diseñado para evaluar si la inmunonutrición enteral perioperatoria (EIN, por sus siglas en inglés) puede reducir las complicaciones postoperatorias o la inflamación en pacientes con cáncer de esófago (CE) sometidos a esofagectomía. Métodos: se realizó una búsqueda sistemática en las bases de datos de PubMed, Embase, Web of Science, EBSCO y Cochrane Library. Se evaluó el efecto de la EIN preoperatoria y/o postoperatoria en un ensayo aleatorizado controlado (RCT) en pacientes con cáncer de esófago sometidos a esofagectomía. Dos investigadores buscaron independientemente artículos, extrajeron datos y evaluaron la calidad de los artículos incluidos. Resultados: el metanálisis incluyó diez ensayos controlados aleatorios en los que participaron 1.052 pacientes, de los cuales 573 fueron incluidos en el grupo EIN y 479, en el grupo de nutrición enteral (NE). En general, no hubo diferencia significativa en la incidencia de neumonía postoperatoria, infección del sitio quirúrgico, absceso intraperitoneal, sepsis e infección del tracto urinario entre los dos grupos. No hubo diferencia significativa en la incidencia de fístula anastomótica postoperatoria, síndrome de distrés respiratorio agudo (SDRA) y mortalidad hospitalaria. Conclusión: la inmunonutrición enteral perioperatoria no puede reducir la incidencia de complicaciones infecciosas postoperatorias y fístulas anastomóticas, ni la PCR postoperatoria ni la IL-6. Pero no aumentó la mortalidad hospitalaria.

The Divergent and Conserved Expression Profile of Turtle Nanog Gene Comparing with Fish and Mammals.

Nanog is a homeodomain-containing transcription factor, and it plays a vital role in maintaining the pluripotency of embryonic stem cells. Nanog's function has been well studied in many species. However, there is lack of reporting on the Nanog gene in reptile. Here, we identified a 1032 bp cDNA sequence of a Nanog gene in Pelidiscus sinensis, known as PsNanog. PsNanog has a highly conserved HD domain and shares a high identity with that of Chelonia mydas and the lowest identity with Oryzias latipes. Similarly, PsNanog presented a tight cluster with C. mydas Nanog, but was far from those of teleosts. Additionally, we cloned a length of 1870 bp PsNanog promoter. Dual luciferase assay showed that the DNA fragment of -1560 to +1 exhibited a high promoter activity. The RT-PCR and RT-qPCR results showed that PsNanog was predominantly expressed in ovary, and then in testis. The in situ hybridization and immunohistochemical analysis showed that PsNanog was expressed in the early primary oocytes and the cytoplasm of the cortical region of stage VIII oocytes in ovary, and distributed in most stages of germ cells in testis. Collectively, the results imply that PsNanog probably has the conserved function in regulating germ cell development across phyla and is also a pluripotent cell gene and expressed in germ cells, which is similar to that in teleosts and mammals.

CKS2 and S100A12: Two Novel Diagnostic Biomarkers for Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a chronic systematicness autoimmunity disease with joint inflammation. RA etiology is still unknown. Early and exact diagnosing is still hard to reach. In the paper, we purposed to discover novel diagnosis biological marker for RA. Two open, usable gene expression profiles of human RA as well as controlled specimens (dataset GSE17755 as well as GSE93272) were downloaded from the GEO database. Differentially expressed genes (DEGs) were screened between 331 RA and 88 control samples. Functional enrichment analysis was applied to explore the possible function of DEGs. Expression levels as well as diagnosis values of biological marker in RA were further verified in our cohort by the use of RT-PCR and ROC assays. We identified 13 DEGs between RA samples and control samples. 13 DEGs were remarkably abundant in NF-kappa B signal pathway. Among the 13 DEGs, CKS2, S100A12, LY96, and ANXA3 exhibited a strong diagnostic ability in screening RA specimens from normal specimens using all AUC > 0.8. Moreover, we confirmed that the expression of CKS2 and S100A12 was distinctly upregulated in RA specimens contrasted to normal specimens. Overall, serum CKS2 and S100A12 could be used as novel diagnosis biological markers for RA patients.

Long non-coding RNA TSPEAR Antisense RNA 2 is downregulated in rheumatoid arthritis and inhibits the apoptosis of fibroblast-like synoviocytes by downregulating microRNA-212-3p (miR-212-3p).

Long non-coding RNA (lncRNA) TSPEAR-AS2 (TSPEAR Antisense RNA 2) participates in many human diseases, while its roles in rheumatoid arthritis (RA) are unknown. Plasma expression levels of TSPEAR-AS2 and microRNA (miR)-212-3p in both RA patients and healthy controls were measured by RT-qPCR. Diagnostic potentials of plasma TSPEAR-AS2 and miR-212-3p were assessed by ROC curve analysis. Normalized expression levels of TSPEAR-AS2 and miR-212-3p were subjected to Pearson's correlation coefficient to evaluate their corrections. TSPEAR-AS2 was significantly downregulated in RA patients, while plasma expression levels of miR-212-3p were significantly increased in RA patients. The expression of TSPEAR-AS2 and miR-212-3p showed promising diagnostic value for RA. Plasma expression levels of TSPEAR-AS2 and miR-212-3p were significantly and inversely correlated in RA patients but not in healthy controls. Besides, overexpression of TSPEAR-AS2 decreased the apoptosis of RA HFLSs, while miR-212-3p increased cell apoptosis. In addition, miR-212-3p attenuated the effects of overexpression of TSPEAR-AS2. Overexpression of TSPEAR-AS2 decreased the expression levels of miR-212-3p in HFLS, while overexpression of miR-212-3p did not affect the expression of TSPEAR-AS2. In conclusion, TSPEAR-AS2 is downregulated in RA and its overexpression can decrease the apoptosis of RA HFLSs by downregulating miR-212-3p.

Changes in rheumatoid arthritis under ultrasound before and after sinomenine injection.

BACKGROUND: Rheumatoid arthritis (RA) is a prevalent clinical autoimmune disease that is commonly treated with diclofenac and methotrexate. In recent years, the application of traditional Chinese medicine in RA has received widespread attention; it promotes blood circulation, strengthens the immune system, and eliminates evil. The sinomenine preparation of Zhingqeng Fengtongning is studied as a possible treatment for patients with RA. AIM: To explore the value of sinomenine injection into the articular cavity for the treatment of RA. METHODS: A total of 94 patients with RA treated from January 2019 to January 2021 were selected and divided into the study and control groups with 47 patients each using a simple random number table method. Both groups received conventional treatment with diclofenac sodium and methotrexate tablets. The control group received diproxone and lidocaine by intra-articular administration while the study group received an intra-articular administration of the sinomenine preparation of Zhengqing Fengning and lidocaine. χ 2 test was used to evaluate the therapeutic effect and synovial thickness, degree of pain through the visual analog scale (VAS), blood flow grade, arthroinflammatory indexes [rheumatoid factor (RF), C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR)] before and after treatment in the two groups. RESULTS: The total effective rate of the study group (93.62%) was higher than that of the control group (78.72%) (P < 0.05). Before treatment, there were no significant differences between the two groups in terms of synovial thickness, VAS score, blood flow grading, levels of RF, and ESR (P > 0.05). After treatment, the synovial thickness and VAS score were significantly lower (P < 0.05) in the study group than in the control group (2.05 ± 0.59 mm vs 2.87 ± 0.64 mm and 2.11 ± 0.62 vs 2.90 ± 0.79 scores, respectively). The rate of blood flow at grade 0 in the study group (76.60%) was higher than that in the control group (57.45%), and the rate of blood flow at grade I (10.64%) was lower than that in the control group (31.91%) (P < 0.05). Furthermore, the levels of RF (55.61 ± 6.13 U/mL), CRP (11.43 ± 3.59 mg/L), and ESR (29.60 ± 5.56 mm/h) in the study group were lower than those in the control group (73.04 ± 9.23 U/mL, 15.07 ± 4.06 mg/L, 36.64 ± 6.10 mm/h, respectively) (P < 0.05). CONCLUSION: Sinomenine administration of Zhengqing Fengtongning in the articular cavity with conventional treatment of RA can improve ultrasonographic blood flow and synovial thickness, reduce pain, regulate inflammation, and enhance therapeutic effect.

Shotgun metagenomics reveals both taxonomic and tryptophan pathway differences of gut microbiota in bipolar disorder with current major depressive episode patients.

BACKGROUND: The microbiome-gut-brain axis, especially the microbial tryptophan biosynthesis and metabolism pathway (MiTBamp), is closely connected to bipolar disorder with current major depressive episode (BPD). METHODS: We performed shotgun metagenomics sequencing (SMS) of faecal samples from 25 BPD patients and 28 healthy controls (HCs). Except for the microbiota taxa and MiTBamp analyses, we also built a classification model using the Random Forests (RF) and Boruta algorithm to find the microbial biomarkers for BPD. RESULTS: Compared to HCs, the phylum Bacteroidetes abundance was significantly reduced, whereas that of the Actinobacteria and Firmicutes were significantly increased in BPD patients. We also identified 38 species increased and 6 species decreased significantly in the BPD group. In the MiTBamp, we identified that two Kyoto Encyclopedia of Genes and Genomes (KEGG) orthologies (KOs) (K00658 and K00837) were significantly lower in the BPD, and five KOs (K01696, K00382, K00626, K01667, and K03781) were significantly higher in the BPD group. We also identified significant genera and species which were closely related to these KOs. Finally, RF classification based on gut microbiota at the genus level can achieve an area under the receiver operating characteristic curve of 0.997. LIMITATIONS: The features of cross-sectional design, limited sample size, the heterogeneity of bipolar disorders, and a lack of serum/plasma tryptophan concentration measurements. CONCLUSIONS: The present findings enable a better understanding of changes in gastrointestinal microbiome and MiTBamp in BPD. Alterations of microbes may have potential as biomarkers for distinguishing the BPD patients form HCs.

MiR-1202 suppresses hepatocellular carcinoma cells migration and invasion by targeting cyclin dependent kinase 14.

Currently, hepatocellular carcinoma (HCC) patients still have poor survival outcomes mainly due to the powerful mobility of HCC cells. Increasing evidences hint that abnormally expressed miRNAs are capable to modulate HCC cells invasion and migration. MiR-1202 has been proposed as a ponderable molecular tumor marker in a variety of tumors. Here, results from real-time PCR indicated the decreased expression of miR-1202 in HCC. Clinically, statistical analysis showed that miR-1202 under-expression was closely associated with metastasis-related clinicopathologic characteristics. In addition, 5-year overall survival (OS) and disease-free survival (DFS) rates of HCC patients with high miR-1202 expression were much better than that in low miR-1202 group. Functionally, gain- and loss-of -function studies were performed to investigate the roles of miR-1202. Intriguingly, Would healing assay and Transwell assays indicated that elevated miR-1202 weakened HCC cells migration and invasion abilities, while miR-1202 knockdown presented the contrary effects. Furthermore, cyclin dependent kinase 14 (CDK14) was identified as a downstream target of miR-1202 by bioinformatics analysis, Dual luciferase reporter assay, detection of CDK14 expression and Pearson correlation analysis. More importantly, rescue experiments demonstrated that CDK14 mediated miR-1202 to exert its anti-tumor effects, which further confirmed the above finding. Taken together, miR-1202 may act as a new biomarker and potential therapeutic target for HCC.

Arsenite inhibits the function of CD133+ CD13+ liver cancer stem cells by reducing PML and Oct4 protein expression.

Cancer stem cells (CSCs) can form new tumors and contribute to post-operative recurrence and metastasis. We showed that CD133+CD13+ hepatocytes isolated from HuH7 cells and primary HCC cells display biochemical and functional characteristics typical of CSCs, suggesting that CD133+CD13+ hepatocytes in primary HCC tumors function as CSCs. We also found that arsenite treatment reduced the viability and stemness of CD133+CD13+ hepatocytes, enhanced the sensitivity of HuH7 cells to pirarubicin, and reduced the tumorigenicity of CD133+CD13+ hepatocytes xenografts in mice. The effects of sodium arsenite treatment in CD133+CD13+ hepatocytes were mediated by the post-transcriptional suppression of PML expression and the inhibition of Oct4, Sox2, and Klf4 expression at the transcriptional level. Incomplete rescue of Oct4 expression in arsenic-treated cells ectopically expressing an siRNA-resistant PML transcript suggested that OCT4 regulation in liver CSCs involves other factors in addition to PML. Our findings provide evidence of a specific role for PML in regulating Oct4 levels in liver CSCs and highlight the clinical importance of arsenic for improving the efficacy of other chemotherapeutic agents and the prevention of post-operative HCC recurrence and metastasis.

Gemcitabine Adjuvant Therapy for Resected Pancreatic Cancer: A Meta-analysis.

Gemcitabine (GEM) is an approved treatment for unresectable pancreatic cancer; however, its role in treating resected pancreatic cancer is less clear. The aim of this study was to investigate the evidence of the role of adjuvant GEM therapy on survival in resected pancreatic cancer. Four phase III randomized trials of adjuvant GEM in patients with resected pancreatic cancer were identified and the hazard ratio (HR) for overall survival were used in this meta-analysis; 2 studies compared GEM treatment with best supportive care and 2 studies with 5-fluorouracil/folinic acid therapy. The pooled data (n=2017 patients) indicated that the overall survival data were homogenous among the studies (Q=4.371; I=31.37%; P=0. 224). The combined HR significantly favors GEM over the other treatments. The overall HR was 0.88 (range, 0. 720 to 0.940; P=0.014). The results indicate that GEM prolongs overall survival compared with other treatments after the resection of pancreatic cancer.

Evaluation of entropy for monitoring the depth of anesthesia compared with bispectral index: a multicenter clinical trial.

BACKGROUND: As a new electroencephalogram (EEG) signal processing technique for monitoring the depth of anesthesia, entropy consists of two indices: reaction entropy (RE) and state entropy (SE). Our study compared entropy with classical bispectral index (BIS) in reduction of myoelectrical interference and noxious stimuli with EEG signals. METHODS: Two hundred and eighty patients (ASA I-II, 18-60 years old) undergoing scheduled surgeries from seven medical centers were enrolled. Anesthesia induction was managed with propofol via the target-controlled infusion (TCI) system. The results of BIS, RE, SE, mean arterial pressure (MAP) and heart rate (HR) were recorded before anesthesia induction, at the moment of unconsciousness, before and 2 minutes after administration of muscle relaxant, and before and one and three minutes after the tracheal intubation. RESULTS: The values of half maximum effective concentrations (EC50), 5% effective concentrations (EC05) and 95% effective concentrations (EC95) of propofol effect-site concentration at the onset of unconsciousness were 1.2 (1.1-1.3 µg/ml), 2.5 (2.4-2.5 µg/ml) and 3.7 (3.7-3.8 µg/ml), while those of the predicted plasma propofol concentration were 2.8 (2.7-2.9 µg/ml), 3.9 (3.8-3.9 µg/ml) and 4.9 (4.8-5.0 µg/ml), respectively. The values of BIS, SE and RE were 62, 59 and 63 when 50% of patients lost consciousness, and 79, 80, 85 and 42, 37, 44, respectively, when 5% and 95% of patients were unconscious. The values of BIS, RE and SE dropped two minutes after the injection of muscle relaxant, but there were no significant differences between RE and SE. MAP and HR increased visibly, which indicated a reaction to tracheal intubation; the values of BIS, RE and SE, however, did not display any significant changes. CONCLUSIONS: This large-sample multicentric study confirmed the values of RE and SE as approximating BIS value, at the onset of unconsciousness during propofol TCI anesthesia. After elimination of myoelectrical activation, all values of RE, SE and BIS decreased significantly and the three indices were less sensitive to noxious stimuli than cardiovascular responses.

Propofol protects against high glucose-induced endothelial dysfunction in human umbilical vein endothelial cells.

BACKGROUND: Hyperglycemia, via peroxynitrite-mediated endothelial nitric oxide synthase (eNOS) enzymatic uncoupling, induced endothelial dysfunction. Propofol has been reported to improve high glucose-induced endothelial dysfunction. However, its mechanisms of action remain unclear. We hypothesized that propofol could improve hyperglycemia-induced endothelial dysfunction by decreasing the peroxynitrite level and thus restoring eNOS coupling. METHODS: At the end of 3 days of incubation in medium with 30 mM glucose, human umbilical vein endothelial cells were treated with different concentrations (0.2, 1, 5, and 25 µM) of propofol for different times (0.5, 1, 2, and 4 hours). In parallel experiments, cells were cultured in 5 mM glucose for 3 days as a control. Nitric oxide (NO) production was measured with a nitrate reductase assay. Superoxide anion (O(2)(·-)) accumulation was measured with the reduction of ferricytochrome c and dihydroethidine fluorescence assay. The treatment that had maximal effect on 30 mM glucose-induced NO production and O(2)(·-) accumulation was applied in the following studies to examine the underlying signaling pathways. eNOS total protein, eNOS dimer and monomer expression, eNOS phosphorylation at Ser(1177), inducible NO synthase total protein, inducible NO synthase dimer and monomer expression, peroxynitrite, and guanosine triphosphate cyclohydrolase I expression were measured by Western blot. Tetrahydrobiopterin (BH(4)) level was measured with liquid chromatography-mass spectrometry. RESULTS: Compared with 5 mM glucose treatment, 30 mM glucose significantly decreased NO production by 60% (P < 0.001) and increased O(2)(·-) accumulation by 175% (P = 0.0026), which were both attenuated by propofol in a concentration- and time-dependent manner. Compared with 5 mM glucose treatment, total eNOS protein expression was increased by 30 mM glucose (P < 0.001), whereas the ratio of eNOS dimer/monomer (P = 0.0001) and eNOS phosphorylation (P < 0.001) were decreased by 30 mM glucose. Propofol did not affect 30 mM glucose-induced total eNOS protein expression, but restored the ratio of eNOS dimer/monomer (P = 0.0005) and increased eNOS phosphorylation (P < 0.001). 30 mM glucose-induced O(2)(·-) accumulation was inhibited by the eNOS inhibitor hydrochloride. Furthermore, compared with 5 mM glucose treatment, 30 mM glucose decreased the BH(4) level (P = 0.0001) and guanosine triphosphate cyclohydrolase I expression (P < 0.001), whereas it increased peroxynitrite level (P = 0.0003), which could all be reversed by propofol (P = 0.0045, P < 0.001, P = 0.0001 vs 30 mM glucose treatment, respectively). CONCLUSIONS: Propofol has beneficial effects on 30 mM glucose-induced NO reduction and O(2)(·-) accumulation in human umbilical vein endothelial cells. This may be mediated through inhibiting peroxynitrite-mediated BH(4) reduction, and restoring eNOS coupling.

Possible involvement of the endocannabinoid system in memory modulation effect of general anesthetics.

Amnesia is a fundamental component of a proper general anesthetic. The mechanism of anesthetic-induced amnesia remains poorly understood. Nowadays, intraoperative awareness and postoperative cognitive dysfunction are two distressing problems receiving increased attention by clinicians, patients and the general public. Extensive evidence indicates that general anesthetics cause amnesia by working on hippocampus and basolateral amygdala (BLA). Recently, evidence from studies in experimental animals has shown that either intra-hippocampus or intra-BLA injection of endogenous cannabinoid receptor 1 (CB1) drugs result in significant changes of cognitive function. In addition, several general anesthetics (i.e. propofol, etomidate and isoflurane) have been reported to interact with the endocannabinoid system. However, there are few studies about whether the CB1 receptor system is involved in anesthetic-induced amnesia. We hypothesize that the CB1 receptor activity in hippocampus and BLA might be regulated by general anesthetics. Once the involvement of the endocannabinoid system in anesthetic-induced amnesia is proved, it will provide a new way to prevent and treat post-traumatic stress disorder caused by intraoperative awareness and postoperative cognitive dysfunction in the future.

Involvement of EphB1 receptor/ephrinB1 ligand in bone cancer pain.

In prior studies, Eph/ephrin system was demonstrated to be involved in inflammatory and neuropathic pain modulation. The present study was to investigate whether the spinal Eph/ephrin signaling was involved in modulation of spinal inflammatory cytokines in bone cancer pain (BCP) of rats. BCP was induced by intra-tibial inoculation of Walker 256 mammary gland carcinoma cells. The expressions of EphB1/ephrinB1 in spinal cord (SC) and dorsal root ganglia (DRG) were determined. At 16 days post inoculation, the pain relieving effect and the mRNA levels of inflammatory cytokines were detected after intrathecal administration of EphB1-Fc (blocker of EphB1 receptor, 10µg). The results showed that the EphB1/ephrinB1 expression was significantly increased in SC, but ephrinB1 was decreased in DRG after Walker 256 inoculation. The mechanical allodynia induced by bone cancer was significantly alleviated by intrathecal administration of EphB1-Fc. Furthermore, the RT-PCR analysis showed that the mRNA levels of IL-1ß, IL-6 and TNF-α were significantly increased at 16 days post Walker 256 inoculation and were significantly suppressed by intrathecal administration of EphB1-Fc in SC. We concluded that Eph/ephrin might be involved in the maintenance of mechanical allodynia, via modulating the expression of spinal inflammatory cytokines, in the present rat model of BCP. This study suggested that Eph/ephrin signaling would be a potential target for the treatment of BCP.

Aspirin and pravastatin reduce lectin-like oxidized low density lipoprotein receptor-1 expression, adhesion molecules and oxidative stress in human coronary artery endothelial cells.

BACKGROUND: Oxidative stress and inflammation are important steps in the pathogenesis of atherosclerosis. We postulated that therapeutic concentrations of aspirin and pravastatin, especially in combination, may suppress oxidative stress and inflammation in endothelial cells, and this concept was examined in human coronary artery endothelial cells (HCAECs). METHODS: Human coronary artery endothelial cells were cultured and treated with oxidized-low density lipoprotein (ox-LDL, 60 microg/ml for 24 hours) alone, or pre-treated with aspirin (1, 2 or 5 mmol/L), pravastatin (1, 5 or 10 micromol/L) or their combination (1 mmol/L aspirin and 5 micromol/L pravastatin), followed by ox-LDL treatment. After respective treatment, superoxide anion production, p38 mitogen activated protein kinase and transcription factor NF-kappaB activation, protein expression of lectin-like ox-LDL receptor-1 (LOX-1) and adhesion molecules, and monocyte adhesion were measured. RESULTS: Ox-LDL treatment greatly elicited its receptor LOX-1 expression, superoxide anion production and inflammatory response, which were minimally affected by low concentration of aspirin (1 mmol/L) or pravastatin (5 micromol/L), but were markedly decreased by their combination. Activation of p38 mitogen activated protein kinase and NF-kappaB, the expression of intercellular adhesion molecule-1 and monocyte chemotactic protein-1, which were only mildly affected by aspirin or pravastatin alone, were significantly attenuated by their combination. As a consequence, monocyte adhesion to endothelial cells was markedly attenuated by the combination of the two agents. Well-known anti-oxidants alpha-tocopherol and gamma-tocopherol had similar inhibitory effects on ox-LDL-mediated oxidative stress and LOX-1 expression as well as monocyte adhesion as did the combination of aspirin and pravastatin. CONCLUSIONS: These studies point to a positive interaction between aspirin and pravastatin with regard to endothelial biology. Anti-oxidant and subsequent anti-inflammatory effect may be one of the potential underling mechanisms.

Effect of thoracic epidural anesthesia with different concentrations of ropivacaine on arterial oxygenation during one-lung ventilation.

BACKGROUND: Thoracic epidural anesthesia can contribute to facilitate the fast-track approach in lung surgery. However, data regarding the effects of thoracic epidural anesthesia on oxygenation during one-lung ventilation (OLV) are scarce and contradictory. Therefore, the authors conducted a prospective, randomized, double-blinded trial in patients undergoing lung surgery under spectral entropy-guided intravenous anesthesia to evaluate the effects of thoracic epidural anesthesia with different concentrations of ropivacaine on oxygenation, shunt fraction (Qs/Qt) during OLV, and maintenance doses of propofol. METHODS: One hundred twenty patients scheduled for lung surgery were randomly divided into four groups to epidurally receive saline (Group S), 0.25% (Group R0.25), 0.50% (Group R0.50), and 0.75% (Group R0.75) ropivacaine. Ropivacaine was administered intraoperatively (6-8 ml of first bolus + 5 ml/h infusion). Arterial oxygen tension (Pao2) and Qs/Qt were measured before, during, and after OLV. RESULTS: Pao2 was significantly lower in Group R0.75 compared with that in Group S and Group R0.25 10 min (170 +/- 61 vs. 229 +/- 68 mmHg, P = 0.01; 170 +/- 61 vs. 223 +/- 70 mmHg, P = 0.03) and 20 min after OLV (146 +/- 52 vs. 199 +/- 68 mmHg, P = 0.009; 146 +/- 52 vs. 192 +/- 67 mmHg, P = 0.03). During OLV, Qs/Qt was significantly higher in Group R0.75 compared with that in Group S and Group R0.25 (P < 0.05). Maintenance doses of propofol were significantly lower in Group R0.75. Vasopressor requirements were higher in Group R0.75. CONCLUSION: A decrease in oxygenation during OLV occurred only at the highest dose of epidural local anesthetic and not at lower doses. Higher doses of epidural medication required less propofol and more vasopressors.

Effects of ropivacaine on adhesion molecule CD11b expression and function in human neutrophils.

Local anesthetics possess a wide range of anti-inflammatory properties through their effects on neutrophils. However, limited information is available on the effects of ropivacaine (a new local anesthetic) on neutrophil function. The aim of this study was to investigate anti-inflammatory properties of ropivacaine with regard to its effects on the expression and function of CD11b in human neutrophils. CD11b expression was examined by flow cytometry and its function was determined by measuring adhesion of neutrophils to human umbilical vein endothelial cells (HUVECs). Ropivacaine inhibited CD11b expression in formyl-methionyl-leucyl-phenylalanine (fMLP)-activated neutrophils in a concentration-dependent manner, but not in a time-dependent manner. The inhibitory potency of ropivacaine was similar to that of bupivacaine and levobupivacaine, but was more potent than that of lidocaine. The up-regulation of CD11b induced by platelet-activating factor (PAF) priming was also inhibited by ropivacaine. fMLP increased adhesion of neutrophils to HUVECs, which was inhibited by ropivacaine. In addition, ropivacaine more potently inhibited the fMLP-induced CD11b expression in the presence of ethylene glycol-bis(2-aminoethylether)-N,N,N ,N -tetraacetic acid (EGTA), a chelator of extracellular Ca(2+). However, ropivacaine showed no effect on the fMLP-induced CD11b expression in the presence of butan-1-ol, a blocker of phospholipase D (PLD) pathway, which completely inhibited the fMLP-induced CD11b expression in neutrophils. Our results suggest that ropivacaine exerts anti-inflammatory activity, and this appears to be mediated through inhibiting the expression and function of adhesion molecule CD11b in neutrophils.

Propofol protects against hydrogen peroxide-induced oxidative stress and cell dysfunction in human umbilical vein endothelial cells.

Propofol has been reported to protect vascular endothelial cells against oxidative stress and dysfunction, but the underlying mechanisms are not clear. In this study, we studied hydrogen peroxide (H(2)O(2))-induced oxidative stress and cell dysfunction in human umbilical vein endothelial cells (HUVECs) and especially, their modulation by propofol. HUVECs were treated with different concentrations (0.1 and 0.5 mM) of H(2)O(2) for different times (1, 3, and 6 h). Then HUVECs were pretreated with different concentrations of propofol (10, 25, and 50 microM), followed by H(2)O(2) treatment (0.5 mM, 3 h). In another set of experiments, we pretreated cells with p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580, followed by H(2)O(2) treatment (0.5 mM, 3 h). After treatment, oxidative stress, p38 MAPK phosphorylation, transcription factor NF-kappaB activation, nitric oxide synthase (NOS) expression, nitric oxide (NO) production, and monocyte adhesion were measured. We observed H(2)O(2) treatment significantly induced oxidative stress, which could be attenuated by 25 microM propofol pretreatment. In addition, H(2)O(2) treatment significantly induced p38 MAPK phosphorylation, NF-kappaB activation, NOS expression, and NO production. More importantly, our study showed these H(2)O(2)-induced changes were attenuated by propofol or SB203580 pretreatment. Further, we measured monocyte adhesion as a marker of endothelial cell dysfunction. H(2)O(2) increased the adhesion of monocytes to HUVECs, and propofol pretreatment reduced the adhesion in a fashion similar to SB203580. We concluded that propofol, by inhibiting p38 MAPK and NF-kappaB activity, decreasing NOS expression, reducing NO production, could protect HUVECs which are exposed to oxidative stress and becoming dysfunctional.