Study on type I interferon and phospho-IRF3 in murine liver dendritic cells after intervened by HBV / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To detect the secretions of type I interferon and the expressions of phospho-IRF3 in murine liver dendritic cells intervened by HBV.</p><p><b>METHODS</b>The murine liver dendritic cells were isolated via anti-CD11c microbeads and were incubated in the presence of GM-CSF and IL-4 to induce the DC generation and proliferation in 24-well cell culture plates. HBV virions were isolated via ultracentrifugation and were detected by quantitative Realtime-PCR. The DCs were divided into two groups: one group was cultured with HBV virions for 24 hours, the other group was cultured without HBV as control group. The cells were harvested at Oh, 1h, 2h, 6h and 24h after being stimulated with poly I:C and the expressions of p-IRF3 and the concentration of IFN beta in supernatants were detected with western blot and ELISA respectively.</p><p><b>RESULTS</b>The IFN beta concentrations at 0 h, 6 h and 24 h in the supernatants of the HBV group and the control group were (12.38 +/- 3.71) pg/ml, (88.67 +/- 9.01) pg/ml and (69.89 +/- 5.80) pg/ml vs (10.83 +/- 4.11) pg/ml, (137.68 +/- 12.28) pg/ml and (72.25 +/- 8.61) pg/ml, respectively. No statistical differences found at 0 h (t = 0.8398, P > 0.05) and 24 h (t = 0.6820, P > 0.05) between the two groups except that at 6 h (t = 9.653, P < 0.01). The expressions of phospho-IRF3 in HBV group were lower than that in control group.</p><p><b>CONCLUSIONS</b>The type I interferon secretion and the phospho-IRF3 expression were decreased in murine liver dendritic cells when intervened by HBV.</p>

Expression of nuclear factor-κB and its regulated cytokines in dendritic cells in patients with chronic severe hepatitis B and its clinical significance / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the correlation of nuclear factor-κB (NF-κB) level and its regulated cytokines in monocyte-derived dendritic cells (MoDC) with illness severity and prognosis in patients with chronic severe hepatitis B (CSHB).</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll density gradient separation from 37 patients with CSHB, 20 chronically HBV-infected patients, and 20 normal controls. Monocytes were acquired using immunomagnetic anti-CD14-beads. Next, monocytes were induced into immature DC (iDC) in vitro. On day six, polyI:C was added to induce DC maturation. Then mature DCs (mDCs) were collected at 48 h after polyI:C treatment to test the expression of NF-κB p50 by real time PCR. The supernatants were also collected respectively. The expression of TNFα and IL-6 in the supernatants were measured by ELISA.</p><p><b>RESULTS</b>The expression of NF-κB p50 in mDCs of patients with CSHB was the highest in three groups (F=19.01, P<0.01). The secretion of TNFα and IL-6 in mDCs from group CSHB was extremely high when compared with the other two groups, the secretions of TNFα in groups CSHB, CHB and NC were(15317.69+/-4124.90) pg/ml, (9670.29+/-3654.68) pg/ml and (6547.43+/-1027.20) pg/ml (F=45.77, P<0.01) respectively, and the productions of IL-6 in groups CSHB, CHB and NC were (1423.78+/-375.14) pg/ml, (862.68+/-93.68) pg/ml and (567.26+/-167.04) pg/ml (F = 67.60, P is less than 0.01), respectively. NF-κB p50 showed significant correlations with TNFα(r=0.52, P<0.01) and IL-6 (r=0.65, P<0.01) in mDCs. Furthermore, the secretions of TNFα and IL-6 in mDCs from group CSHB were negatively associated with PTA (r=-0.41, P=0.01; r=-0.40, P=0.01), but not with ALT, TBil and virus loads (r=-0.03, P=0.85, r=0.01, P=0.93, r=0.01, P=0.95; r=-0.09, P=0.58, r=0.16, P=0.34, r=0.09, P=0.59). No significant difference found in the expression of TNFα and IL-6 between the survival subgroup and the death subgroup in group CSHB (t=0.42, P=0.67; t=0.76, P=0.45).</p><p><b>CONCLUSIONS</b>The expression levels of NF-κB and its regulated cytokines in monocyte-derived DCs in patients with chronic severe hepatitis B were up-regulated and perhaps associated positively with the illness severity.</p>

Phenotypes and functions of dendritic cells derived from peripheral blood monocytes of chronic hepatitis B patients with different HBV DNA loads / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the phenotypes and functions of peripheral blood monocyte derived dendritic cells (DC) of chronic hepatitis B (CHB) patients with different HBV DNA loads.</p><p><b>METHODS</b>Twenty-eight CHB patients were included in this study. All patients were treated with nucleoside analogues (lamivudine or LdT or adefovir) for 24 weeks. Peripheral blood HBV DNA loads and liver biopsies were assessed before and after the treatment. The patients were divided into two groups according to their peripheral blood HBV DNA loads: a high-load group with HBV DNA loads higher than 10(5) copies/ml, and a low-load group with HBV DNA loads lower than 10(3) copies/ml. Ten healthy people were included as controls. Peripheral blood DC of each subject was enriched. The phenotypes of DC were subjected to flow cytometric analysis. The lymphocyte allo-stimulatory capacity of DC was evaluated through MTT assay. IL-10 and IL-12 production were quantified by ELISA.</p><p><b>RESULTS</b>DC proliferated successfully when stimulated by cytokines in vitro; however, DC of the CHB patients proliferated much slower than those of the healthy controls. The expression of DC surface molecules such as HLA-DR, CD86, CD80 and CD83 had a positive rate of over 80% in the normal population. However in our CHB patients they showed lower than normal expressions, especially the HLA-DR, CD86, CD80 and CD83, but the differences were not significant between the two groups with different virus loads. The stimulatory capacity of the DC in mixed lymphocyte reaction showed no difference between the two groups of patients, but both were lower than that of the healthy controls. The production of IL-12 and IL-10 also decreased significantly in the patients.</p><p><b>CONCLUSIONS</b>Peripheral DC of CHB patients have some defects in their phenotypes and their stimulatory capacity. The changes in phenotypes and down-regulation of the functions are not relevant to peripheral HBV DNA loads of the patients.</p>

Effect of fresh gas flow on isoflurane pharmacokinetics during anesthesia induction / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of different fresh gas flow (FGF) rates on isoflurane pharmacokinetics during anesthesia induction.</p><p><b>METHODS</b>Sixty female patients (ASA class I-II, age range of 18-49 years) scheduled for gynecological laparoscopic surgery were randomly divided into groups I, II, and III (n=20) for isoflurane inhalation with FGF rate of 1, 2, and 3 L/min, respectively. Each group was further divided into two equal subgroups according to the setting concentration of the isoflurane vaporizer at 1% (groups I 1, II 1, and III 1) and 2% (groups I 2, II 2, and III 2). Isoflurane with different setting concentrations was administered under different FGFs in the patients after tracheal intubation following anesthesia induction, and the inspiratory concentration (CIiso) and expiratory concentration (CEiso) of isoflurane in the airway were monitored and recorded every 3 min for totalling 18 min, with the observation time points marked as T1 to T6, respectively.</p><p><b>RESULTS</b>CIiso and CEiso varied significantly at different time points and between different subgroups (P<0.05). In each subgroup, CIiso and CEiso increased along with time and reached a relatively stable stage at 9 min, but failed to reach the setting concentration during the observation period. At different observation time points, CIiso and CEiso in the subgroups with setting isoflurane concentration of 2% were almost twice as much as that in the subgroups with setting isoflurane concentration of 1%.</p><p><b>CONCLUSIONS</b>CIiso and CEiso increase along with time lapse in all the groups and reach a relatively stable stage at 9 min after inhalation initiation, but can not reach the setting concentration. The larger the FGF and setting concentration, the faster CIiso and CEiso increase.</p>

Study on the expression of Toll like receptor 3 on dendritic cells derived from peripheral blood monocyte of chronic hepatitis B patients / 中华传染病杂志

Objective To investigate the expression of Toll-like receptor 3 (TLR3) on dendritic cells(DCs) derived from peripheral blood mononuclear cells(PBMCs) of chronic hepatitis B(CHB) patients and to explore the mechanism of sustained infection of hepatitis B virus (HBV). Methods Twenty CHB patients were randomly screened in the study,and ten healthy persons were recruited as controls.The monocytes isolated from peripheral blood of candidates were incubated with recombinant human granuloeyte macrophage colony-stimulating factor (rhGM-CSF) and rhIL-4 to induce the DCs generation and proliferation.Then the phenotype of DCs was identified by micro- scope.The expressions of the phenotypes[histocompatibility leukocyte antigen(HLA)-DR,CD80, CD86,CD83]of immature and mature DCs were measured by flow cytometer.Furthermore,the ex- pression of TLR3 on mature DCs(mDC) and immature DCs(imDC) was determined by flow cytometry and Western blot analysis respectively.Results As for healthy volunteers,the expressions of CD80, CD86,HLA-DR and CD83 on DCs at the 7th day,which were(82.35?8.67)%,(79.61?10.08)%, (92.79?8.48)% and (83.76?5.47)% respectively,were significantly higher than those at the 5th day which were(28.31?8.79) %,(31.17?11.23)%,(27.61?10.28)% and (23.46?11.53)% respec- tively(P0.05).The expression of TLR3 on imDC was significantly higher than that on mDC at control group (P0.05).And the expression of TLR3 on imDC in CHB patients group was significantly lower than that of control group(P

Effects of different fresh gas flow on pharmacodynamics of isoflurane during anesthesia induction / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of different fresh gas flows (FGFs) on the pharmacodynamics of isoflurane during anesthesia induction.</p><p><b>METHODS</b>Sixty female ASA class I or II patients (aged from 18 to 49 years) scheduled for gynecologic laparoscopic surgery were randomly divided into groups I, II, and III (n=20). The FGFs for group I, II, and III was 1, 2 and 3 L/min, respectively, and each group was further divided into two equal subgroups according to the setting concentrations of isoflurane vaporizer (Co), which was 1% in groups I1, II1, and III1 and 2% in groups I2, II2, and III2. Isoflurane at different setting concentration was administered under different FGF in the patients after tracheal intubation following anesthesia induction. The systolic blood pressure (SBP), diastolic blood pressure (DBP), main arterial blood pressure (MAP), heart rate (HR) and bispectral index (BIS) were recorded before anesthesia induction and every 3 min after tracheal intubation. Patients given ephedrine and atropine were also recorded. The patients' consciousness during anesthesia were followed up and recorded. The inspiratory concentration (CIiso) and expiratory concentration (CEiso) of isoflurane in the airway were monitored and recorded every 3 min. The observation after intubation lasted for 18 min, during which stimulation of the patients was avoided, and the operation began after the observation.</p><p><b>RESULTS</b>There was a close correlation between BIS and CIiso and between BIS and CEiso (r=-0.904 and -0.893, respectively). The incidence of hypotension was significantly different between groups III and I (P<0.01), and between the subgroups in groups II and I (P<0.05). No bradycardia occurred and no consciousness reported awareness during anesthesia.</p><p><b>CONCLUSIONS</b>Between the completion of tracheal intubation and beginning of the surgery, 1% or 2% Co under a moderate FGF (1-3 L/min) may guarantee the patients' unconsciousness, but hypotension is less likely under a relatively low flow (1-2 L/min) than a higher flow (3 L/min). Higher FGF and Co result in faster induction of deep anesthesia and higher incidence of hypotension.</p>

Clinical analysis of 146 patients with cryptococcal meningitis / 中华传染病杂志

Objectives To investigate the clinical features,prognosis and risk factors of patients with cryptococcal meningitis.Methods Totally 146 patients with cryptococcal meningitis who were hospitalized in Huashan Hospital from January 2000 to December 2006 were enrolled in this study.The clinical data including diagnosis and misdiagnosis,experimental and etiology tests,treat- ments and prognosis from all the patients were analyzed retrospectively.Results Among the 146 patients enrolled in the study,78 patients(53.4%)had concomitant diseases.The misdiagnosis rate of all patients was 72.6%(106/146).The positive rate of cerebrospinal fluid(CSF)India ink smear was 59.6%(87/106),while 43.2%(63/146)cases of cryptococcus neoformans culture in CSF was positive.The positive rate of Latex agglutination test(LAT)was 91.7%(134/146)in CSF among all patients.The treatments were as follows:combination of Amphotericin B(AmpB)or its lipid formula- tions with flucytosine(5-FC)(98 cases),including combination with Fluconazole initally(62 cases), single therapy of Fluconazole(13 cases).Ommaya implanted for lateral cerebral ventricle drainage(53 cases)and AmpB intrathecal injection(53 cases).The average dose of AmpB is 3.06 g.The course of treatment lasted from 12 weeks to 20 months.There were 104 patients(71.2%)cured,27(18.5%) improved,15(10.3%)died and 34(23.3%)relapsed.Conclusions High misdiagnosis rate is common in patients with cryptococcal meningitis.Immunodeficiency is the major risk factor for cryp- tococcal meningitis.CSF LAT is the most sensitive diagnostic test.Early diagnosis,combination of AmpB with 5-FC antifungal therapy and control of acute intracranial hypertension are the keys to im- prove prognosis of cryptococcal meningitis.