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Rickettsiae are obligate intracellular bacteria that can cause life-threatening illnesses. There is an ongoing debate as to whether established infections by one

Background Spermine is frequently elevated in tumor tissues and body fluids of cancer patients and is critical for cancer cell proliferation, migration and invasion. However, the immune functions of spermine in hepatocellular carcinoma progression remains unknown. In the present study, we aimed to elucidate immunosuppressive role of spermine in hepatocellular carcinoma and to explore the underlying mechanism. Methods Whole-blood spermine concentration was measured using HPLC. Human primary HCC tissues were collected to examine the expression of CaSR, p-Akt, β-catenin, STT3A, PD-L1, and CD8. Mouse model of tumorigenesis and lung metastasis were established to evaluate the effects of spermine on hepatocellular carcinoma. Western blotting, immunofluorescence, real time PCR, digital Ca2+ imaging, and chromatin immunoprecipitation assay were used to investigate the underlying mechanisms by which spermine regulates PD-L1 expression and glycosylation in hepatocellular carcinoma cells. Results Blood spermine concentration in the HCC patient group was significantly higher than that in the normal population group. Spermine could facilitate tumor progression through inducing PD-L1 expression and decreasing the CD8+ T cell infiltration in HCC. Mechanistically, spermine activates calcium-sensing receptor (CaSR) to trigger Ca2+ entry and thereby promote Akt-dependent β-catenin stabilization and nuclear translocation. Nuclear β-catenin induced by spermine then activates transcriptional expression of PD-L1 and N-glycosyltransferase STT3A, while STT3A in turn increases the stability of PD-L1 through inducing PD-L1 protein N-glycosylation in HCC cells. Conclusions This study reveals the crucial function of spermine in establishing immune privilege by increasing the expression and N-glycosylation of PD-L1, providing a potential strategy for the treatment of hepatocellular carcinoma.

Objective Neuraxial block is the most common anesthesia method for cesarean section (CS). However, for some urgent and high-risk cesarean delivery, general anesthesia (GA) also plays a very important role. We aimed to find out the reasons of choosing GA for CS in our center and the factors that may be related to the maternal and fetal outcomes.Methods We retrospectively selected parturients who had CS procedures under GA in Peking Union Medical College Hospital from January 1, 2014 to December 31, 2016. Clinical data (baseline maternal status, preoperative status, perioperative information, maternal and fetal outcomes) of parturients and neonates were collected and analyzed. We summarized the common reasons for applying general anesthesia, and compared the back-to-ICU ratio and hospital stay time between parturients with different maternal American Society of Anesthesiologists (ASA) grade, gestational weeks and intraoperative blood loss, as well as the fetal one-minute Apgar score between different maternal ASA grade and gestational weeks.Results There were 98 cases of CS under GA enrolled in the study. Among the maternal and fetal factors, pregnancy with internal or surgical diseases is the most common reason (59 cases, 60.2%) for choosing GA, followed by the placenta and fetal membrane abnormalities (38 cases, 38.8%) and the pregnancy-specific disorders (36 cases, 36.7%). ASA grade Ⅲ-Ⅳ of parturients (χ 2=44.3, P 800 ml (χ 2=5.5, P

Anti-N-methyl-D-aspartate receptor (anti-NMDA-R) encephalitis is a newly recognized neuro-autoimmune disease. Patients usually present with a series of neurological and psychiatric syndromes including memory impairment, seizures, dyskinesia, autonomic nervous system dysfunction, etc. Anti-NMDA-R encephalitis usually affects young women with teratoma being a common co-existing condition in nearly 47% of the diagnosed patients. Previous studies have shown that teratoma removal, being the current first-line therapy, may enhance the effectiveness of autoimmune therapy and decrease the incidence of relapse.[1] However, poorly controlled encephalitis poses as a perioperative anesthetic risk. Therefore, timing of the surgery is of great concern and is a question yet to be answered. Due to low incidence of anti-NMDA-R encephalitis in general population, literature of the perioperative management in these group of patients is scarce. Only few case reports were found yet, none has discussed the issue of surgery timing. In this study, 6 patients with anti-NMDA-R encephalitis undergoing teratoma removal were reviewed, and timing of surgery and anesthetic management for these patients were discussed. A computerized search of the Medical Records Database was conducted from June 2011 to May 2014 to identify patients with anti-NMDA-R encephalitis and ovarian teratoma. Approval from Institutional Review Board of Peking Union Medical College Hospital (PUMCH) was obtained. Patients who underwent tumor removal under general anesthesia were included. Data record include age, sex, clinical presentations, laboratory findings, diagnostic modalities (electroencephalogram [EEG], cranial magnetic resonance imaging [MRI], serum/cerebrospinal fluid [CSF] autoantibody test, and abdominal ultrasound), preoperation immunotherapy (drugs and dosage) and status of disease control, anesthesia course (anesthetic and emergence), and postoperative course (postoperative mechanical ventilation time, Intensive Care Unit [ICU] length of stay [LOS], and postoperative LOS). Diagnosis of anti-NMDA-R encephalitis as well as evaluating status of disease was double confirmed by a senior neurologist from PUMCH specializing in autoimmune encephalitis. Six female patients with anti-NMDA-R encephalitis were identified from the electronic database. They were numbered as Patient 1 to Patient 6 randomly. The age of the six patients was 22, 26, 28, 32, 10, and 30 years, respectively. They were generally in good health otherwise from teratoma and encephalitis. All six patients were diagnosed with anti-NMDA-R encephalitis by the combination of clinical presentation, EEG, MRI, and serum/CSF anti-NMDA-R antibody test. All the patients present with psychiatric symptoms, seizures, and disturbances of consciousness during the course of disease; four (Patient 1, 3, 4, and 5) had autonomic instability including arrhythmia (sinus tachycardia or sinus bradycardia) and central hypoventilation. Before the operation, three patients (Patient 1, 2, and 3) received intravenous immunoglobulin (IVIg), two patients (Patient 4, 5) received IVIg and corticosteroids. One patient (Patient 6) received IVIg, corticosteroids, plasma exchange therapy, as well as cyclophosphamide treatment. After immunotherapy, two patients (Patient 4, 6) experienced near-complete recovery and other four patients still have psychiatric symptoms and seizures. One patient was transferred to ICU with cardio-respiratory support due to respiratory insufficiency [Table 1]. Table 1 Autoimmune therapy, status of preoperative symptom control and postoperative recovery of the six patients All six patients underwent laparoscopic ovarian cystectomy under general anesthesia. For the two near-complete-recovery patients, induction was carried out with propofol, morphine, and cisatracurium for Patient 4 and propofol, midazolam, fentanyl, and rocuronium for Patient 6. Anesthesia was maintained with targeted controlled infusion of propofol (effect-site concentration of 3–4 μg/ml) + 1:1 air in oxygen for Patient 4, and sevoflurane + 1:1 N2O in oxygen for Patient 6. Both of the patients emerged from anesthesia uneventfully and were extubated right after the operation. They were sent back to general ward. No anesthesia-related complications or exacerbation of neurological symptoms were noted after the surgery. For the rest four patients whose symptoms were not well-controlled, anesthesia induction was with propofol, fentanyl, and rocuronium. Anesthesia maintenance was with sevoflurane + 1:1 air in oxygen for Patient 3, 5 and with sevoflurane + 1:1 N2O in oxygen for Patient 1, 2. All of them were transferred to ICU with tracheal intubation. Patient 3 was extubated 1-day after surgery without anesthesia-related complications. Patient 1, 2, and 5 were kept intubated after surgery for 46, 4, and 21 days, respectively, and all of them developed ventilator-associated pneumonia or pulmonary infection due to prolonged intubation. Two patients (Patient 1, 5) received tracheotomy and Patient 1 underwent unexpected tube prolapse due to dysphoria. Patients who got pneumonia recovered after treatment with antibiotics and the tracheostoma healed up before discharge [Table 2]. Table 2 Anesthetic management for the six patients with anti-NMDA-R encephalitis Postoperative ventilation time, ICU LOS, and postoperative LOS were longer in the patients whose symptoms were poorly controlled after autoimmune therapy (Patient 1, 2, and 5) than well-controlled ones (Patient 4, 6). Management of anti-NMDA-R encephalitis mainly involves immunotherapy and removal of the teratoma if present together. Early tumor removal was strongly advocated for patients with anti-NMDA-R encephalitis by some authors,[1] but the appropriate timing for surgery is still unknown. This study highlights the importance of adequate immunotherapy before surgery. Patients with poor disease control had longer postoperative invasive ventilation time, ICU LOS, postoperative LOS, or higher incidence of pulmonary complications while patients with better disease control had a smoother recovery. Therefore, it is assumed that although tumor removal may enhance the effectiveness of drug therapy, inadequate preoperative therapy may prolong postoperative ventilation time and result in unwanted pulmonary complications. This suggests that it may be appropriate to provide adequate immunotherapy for patients with anti-NMDA-R encephalitis before the surgery. Patient 1 and Patient 5 share similar status of preoperative disease control, but Patient 5 who received both IVIG and methylprednisolone led a relatively better recovery than Patient 1 who received only IVIG. This suggested that intensive combined treatment might be beneficial to patients with severe anti-NMDA-R encephalitis. Many anesthetic drugs can inhibit NMDA-R, and may induce the same symptoms as those observed in anti-NMDA-R encephalitis. NMDA-R can be strongly inhibited by nitrous oxide and ketamine. Therefore, it is presumed that patients with anti-NMDA-R encephalitis may be more sensitive to ketamine and nitrous oxide, which may aggravate the symptoms of encephalitis transiently during or after the operation. As a consequence, most authors suggested that ketamine and nitrous oxide should be avoided during the anesthesia.[2] But in our cases, nitrous oxide had been used in three patients. The effect of nitrous oxide on encephalitis could not be accessed due to continued sedation in ICU. In one patient whose symptom had been well-controlled before surgery thus was transferred to general ward without further sedation, no symptom deterioration was noted after anesthesia. Inhalational anesthetics are strong inhibitors of NMDA-R but not specific. They can also act on other ligand-gated ion channels such as GABAA and glycine receptors. Propofol seems to produce general anesthesia mainly by enhancing GABA pathway.[2] But a recent in vitro study showed that propofol may also block NMDA-R at the concentrations routinely used in general anesthesia[3] although the clinical relevance of this inhibition is not established. Therefore, the clinical effect of inhalational anesthetics and propofol on anti-NMDA-R encephalitis is still eluding. However, Lapebie et al.[4] did report a case of symptom deterioration after general anesthesia with propofol and sevoflurane. Splinter and Eipe[5] reported pronounced hypotension secondary to propofol injection at induction of anesthesia for diagnostic lumbar puncture. These suggest that patients with anti-NMDA-R antibody may be sensitive to propofol and sevoflurane. Interestingly, Splinter and Eipe[5] also reported that another lumber puncture was performed several months later for the same patient with propofol and no hypotension happened. In this study, propofol and sevoflurane were separately used for two patients whose symptoms had been well-controlled. Both of them were extubated immediately and emerged uneventfully after the surgery. Neither hypotension nor symptom deterioration had been found. So, we hypothesize that patients without disease control may be sensitive to anesthesia agents such as sevoflurane and propofol. While in patients whose preoperative symptom is under control, it may be safer to use sevoflurane, propofol, and nitrous oxide. But further investigation with more cases is needed to validate our findings. Our study shows that adequate preoperative immunotherapy and good disease control may be critical to anesthetic and postoperative management of patients with anti-NMDA-R encephalitis. Limited data show that commonly used anesthetics such as propofol, sevoflurane, and N2O may be safe for patients whose encephalitis had been well-controlled with immunotherapy. Financial support and sponsorship Nil. Conflicts of interest There are no conflicts of interest.

Oral Diseases (2010) 16, 343–350 Objective: Toll-like receptors (TLRs) are the essential components in the innate and adaptive immune systems. Colony stimulating factor 2 (CSF2) is a cytokine that may prevent endotoxin tolerance, and LY64 has the ability to interfere with the recognition of bacteria via TLR4. The aim of this study was to explore the in vivo expressions of TLR2, TLR4, CSF2 and LY64 in Chinese chronic periodontitis patients. Methods: Gingival biopsies were collected from 24 chronic periodontitis patients and 19 healthy controls. The gene expression profiles of TLR2, TLR4, CSF2 and LY64 were investigated by real-time polymerase chain reaction, and the protein expressions of TLR2 and TLR4 were detected by immunohistochemistry. In addition, the levels of CSF2 in gingival crevicular fluid (GCF) were determined by ELISA. Results: The higher mRNA expressions of TLR2, TLR4 and CSF2, and the lower mRNA expression of LY64 were detected in chronic periodontitis patients. And the increased protein expressions of TLR2 and TLR4 were confirmed by immunohistochemistry. In addition, the increase of total amount of CSF2 in GCF was observed in chronic periodontitis patients. Conclusions: Our results suggest that TLR2 and TLR4 may play a role in periodontal pathogenesis. In addition, CSF2 and LY64 may contribute to the regulation of inflammatory response and maintaining periodontal homeostasis.

To investigate the antitumor lymphangiogenesis activity and the synergistic effect of indomethacin and oxaliplatin in a transplantation tumor model of human lung cancer in nude mice.Human lung cancer A549 cells were injected subcutaneously into BALB/c nude mice. The mice were randomly divided into 4 groups: the control group, the indomethacin group, the oxaliplatin group, and the indomethacin combined with oxaliplatin group. The tumor volume was measured in a 7 day interval, and the mice were sacrificed after 42 days. The tumor tissues were collected and prepared for detection by immunohistochemistry of expressions of VEGF-C, Survivin, β-catenin and intratumoral microlymphatic density(MLD). Real-time fluorescence quantitative RT-PCR was performed for detecting the expression of VEGF-C and Survivin mRNA. SPSS version 16.0 was used for statistical analysis. Data were present as (x(-) ± s), and means were compared with analysis of variance. The correlation of 2 variables was analyzed by liner correlation analysis.The tumor volumes in the indomethacin group, oxaliplatin group, and indomethacin plus oxaliplatin group [(1322 ± 327) mm(3), (962 ± 221) mm(3), (611 ± 161) mm(3)] decreased significantly as compared with that of the control group [(1664 ± 318) mm(3), F = 23.331, P0.01]. Compared with the control group, the expression levels of VEGF-C, Survivin, β-catenin and MLD protein in the indomethacin group, and indomethacin plus oxaliplatin group were significantly reduced (P0.05). Immunohistochemical studies showed an increase in the expression of VEGF-C in the oxaliplatin group (20 825 ± 2067) as compared to the control group (16 075 ± 875, F = 97.24, P0.05), but no significant differences in β-catenin and MLD (17 396 ± 1693, 9666 ± 978) as compared with the control group (9824 ± 1181, 17 588 ± 1698)respectively, P0.05. VEGF-C and Survivin proteins were positively correlated with MLD respectively (t = 0.737, 0.662, respectively P0.01), and β-catenin protein was positively correlated with Survivin protein (r = 0.582, P0.01). VEGF-CmRNA was positively correlated with its protein (r = 0.873, P0.01).The combined use of indomethacin and oxaliplatin promoted antitumor lymphangiogenesis activity, possible by inhibiting the VEGF-C/VEGFR-3 and Wnt pathways.

Periodontitis is a bacterially induced chronic inflammatory disease. Toll-like receptors (TLRs), which could recognize microbial pathogens, are important components in the innate and adaptive immune systems. Both qualitatively and quantitatively distinct immune responses might result from different bacteria stimulation and the triggering of different TLRs. This study explores the interaction of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) with TLR2 and TLR4.We studied the gene expression changes of TLR2 and TLR4 and cytokine production (interleukin-1β, -6, -8, -10, and tumor necrosis factor-alpha) in human periodontal ligament cells (HPDLCs) stimulated with heat-killed bacteria or P. gingivalis lipopolysaccharide (LPS) in the presence or absence of monoclonal antibodies to TLR2 or TLR4 (anti-TLR2/4 mAb).Both test bacteria and 10 microg/ml P. gingivalis LPS treatment increased the gene expression of TLR2 and TLR4 and cytokine production in HPDLCs. In addition, these upregulations could be blocked by anti-TLR2/4 mAb. However, the expression of TLR4 mRNA in HPDLCs stimulated with 1 microg/ml P. gingivalis LPS was not increased. No differences were found in the cytokine production caused by 1 microg/ml P. gingivalis LPS treatment in the presence or absence of anti-TLR4 mAb.These patterns of gene expression and cytokine production indicate that Gram-negative periodontal bacteria or their LPS might play a role in triggering TLR2 and/or TLR4, and be of importance for the immune responses in periodontitis.

Indolizine and annulated indolizine derivatives incorporating a cyclopropylcarbonyl group were synthesized in a one pot procedure by the tanden reactions of [3+2] cycloaddition of the corresponding N-ylide with electron deficient alkene. Seventeen indolizine derivatives were reported for the first time. All the compounds were examined for their antiproliferative activity against the human hepatocellular liver carcinoma (Hep-G2) cell line by MTT method. Among the compounds tested, 5a, 5d, 5 g and 5 j showed the most favorable activities with IC(50) values of 0.39, 0.48, 0.29 and 0.20 microg/mL. Especially, compound 5 j displayed potent antiproliferative activities with IC(50) value of 0.20 microg/mL, and showed significant EGFR kinase inhibitory activity with IC(50) value of 0.085 microM. Docking simulations of 5 j were carried out to illustrate the binding mode of the molecular into the EGFR active site.

Introduction: The aim of this study was to investigate the prevalence and molecular diversity of Archaea in the subgingival crevices of patients with chronic periodontitis. Methods: Subgingival plaque was collected from 41 patients with chronic periodontitis and 15 healthy subjects. The prevalence of Archaea in those plaque samples was tested by polymerase chain reaction with two broad-range archaeal primer sets. Amplicons from eight Archaea-positive plaque samples were cloned and sequenced for molecular diversity analysis using one of these two primer sets and a novel third primer set. Results: Archaea were detected in the subgingival plaque of patients with chronic periodontitis at a prevalence of 70.7–73.2%, but were not detected in healthy subjects. Using one primer set, all sequences of the archaeal amplicons were identified as Methanobrevibacter oralis-like species. With another primer set, the amplicons were also found to be identical to the uncultured M. oralis-like species except one phylotype was found to belong to the class Thermoplasmata. Conclusion: Archaea might be correlated with periodontal diseases. The diversity of Archaea associated with periodontitis was limited. Almost all sequenced amplicons fell into the genus Methanobrevibacter of the Euryarcheota phylum. M. oralis-like species was the predominant but non-exclusive archaeon in the subgingival dental plaque of patients with periodontitis.

Toll-like receptors (TLRs) are a group of sensors on the surface of antigen-presenting cells, such as dendritic cells and macrophages, which recognize microbial pathogens and induce innate and adaptive immune responses. Periodontitis is an inflammatory disease characterized by the destruction of tooth-supporting structures. In order to address whether TLR4 signaling plays a role in periodontitis, we studied the gene expression change in human periodontal ligament cells (HPDLCs) in response to TLR4 ligand, lipopolysaccharide treatment by microarray analysis. Expression of TLR4 was detected in HPDLCs. Lipopolysaccharide treatment increased the expression of 12 genes (more than twofold), including TLR4, TLR5, TLR7, Pellino 1, colony stimulating factor 2 (CSF2) and IL-6. In addition, the expression of 15 genes (less than equal to twofold) was decreased, including Fos, LY64 and LY86. In addition, real-time PCR was used to confirm the change of gene expression of TLR4, IL-6 and Fos. We also showed that the upregulation of IL-6 by lipopolysaccharide treatment was TLR4-dependent. This pattern of gene expression indicates that pathogens may trigger TLR4 signaling and cause periodontitis. Manipulating TLR4 signaling may potentially become one of the recognized therapies for periodontitis.

To evaluate the effect of TLR4 on expressions of p-IRAK1 and p-IkappaB-alpha in HPDLCs stimulated by lipopolysaccharide.Western blot and image analysis were used to detect the levels of p-IRAK1 and p-IkappaB-alpha in HPDLCs stimulated by 1 microg/ml LPS after 2.5,5,10 and 15 minutes, and to detect the levels of these proteins in HPDLCs stimulated by 1 microg/ml LPS after pretreatment with monoclonal antibody of TLR4. Statistical analysis was performed with SPSS10.0 software package for one-way ANOVA.The expressions of p-IRAK1 and p-IkappaB-alpha in HPDLCs were enhanced after stimulation of LPS. The gray scale values of protein band at 0 minute were 0.054, 0.19 respectively, significantly increased to 0.785, 0.809 after 5 minutes (P0.05). After pretreatment with monoclonal antibody of TLR4, the expressions of these proteins weakened compared with those without pretreatment. The gray scale values of protein band significantly decreased from 0.82,0.874 to 0.099,0.201, respectively (P0.05).TLR4 is involved in the signaling transduction in HPDLCs stimulated with LPS, and might be involved in the progression of periodontis.