Effect of electroacupuncture stimulation of "Sibai"(ST2) and "Quanliao"(SI18) on excitatory neurons of sensorimotor cortex in mice. / 电针"四白"和"颧髎"æ¿€æ´»å°é¼ èº¯ä½“æ„Ÿè§‰è¿åŠ¨çš®å±‚çš„å ´å¥‹æ€§ç¥žç»å ƒ.

OBJECTIVES: To observe the activation state and neuronal types of somatosensory cortex and the primary motor cortex induced by electroacupuncture (EA) stimulation of "Sibai" (ST2) and "Quanliao" (SI18) acupoints in mice. METHODS: Male C57BL/6J mice were randomly divided into blank control and EA groups, with 6 mice in each group. Rats of the EA group received EA stimulation (2 Hz, 0.6 mA) at ST2 and SI18 for 30 minutes. Samples were collected after EA intervention, and immunofluorescence staining was performed to quantify the expression of the c-Fos gene (proportion of c-Fos positive cells) in the somatosensory cortex and primary motor cortex. The co-labelled cells of calcium/calmodulin-dependent protein kinase Ⅱ (CaMKⅡ) and gamma-aminobutyric acid (GABA) in the somatosensory cortex and primary motor cortex were observed and counted by using microscope after immunofluorescence staining. Another 10 mice were used to detect the calcium activity of excitatory neurons in the somatosensory cortex and primary motor cortex by fiber photometry. RESULTS: In comparison with the blank control group, the number of c-Fos positive cells, and the proportion of c-Fos and CaMKⅡ co-labelled cells in both the somatosensory cortex and primary motor cortex were significantly increased after EA stimulation (P<0.05). No significant changes were found in the proportion of c-Fos and GABA co-labeled cells in both the somatosensory cortex and primary motor cortex after EA. Results of fiber optic calcium imaging technology showed that the spontaneous calcium activity of excitatory neurons in both somatosensory cortex and primary motor cortex were obviously increased during EA compared with that before EA (P<0.01), and strikingly reduced after cessation of EA compared with that during EA (P<0.05). CONCLUSIONS: Under physiological conditions, EA of ST2 and SI18 can effectively activate excitatory neurons in the somatosensory cortex and primary motor cortex.

microRNA-1-3p and T-synthase mRNA have high diagnostic efficacy on intestinal mucosal barrier dysfunction in patients with severe acute pancreatitis.

Acute pancreatitis (AP) is an inflammatory disorder of the pancreas that can be complicated by intestinal mucosal barrier dysfunction (SAP&IBD). The current study sought to examine the diagnostic efficacy of miR-1-3p and T-synthase mRNA in SAP&IBD patients. First, SAP patients were assigned to SAP&IBD and SAP groups. Serum miR-1-3p expression and T-synthase mRNA expression patterns in peripheral blood B lymphocytes were measured using RT-qPCR. Pearson tests, ROC curve analysis, and multivariate logistic regression were used to analyze the correlation between miR-1-3p/T-synthase mRNA and clinical data, their diagnostic efficiency, and independent risk factors for SAP&IBD patients, respectively. The results showed that serum miR-1-3p in the SAP&IBD group was elevated, and T-synthase mRNA expression in peripheral blood B lymphocytes was diminished. Additionally, serum miR-1-3p expression in SAP&IBD patients was negatively correlated with T-synthase mRNA expression, and positively correlated with their Ranson score, CRP, IL-6, DAO, and D-Lactate levels. Meanwhile, T-synthase mRNA level was negatively correlated with IL-6, DAO, and D-Lactate levels. Both, serum miR-1-3p, T-synthase mRNA, and their combination were found to exhibit diagnostic efficiency for SAP&IBD patients, and were independently associated with IBD in SAP patients. Collectively, our findings suggest that miR-1-3p and T-synthase serve as independent risk factors for SAP&IBD patients and can aid the diagnosis of IBD in SAP patients.

miR-155 facilitates calcium oxalate crystal-induced HK-2 cell injury via targeting PI3K associated autophagy.

Nephrolithiasis is one of the most common and highly recurrent diseases worldwide. Accumulating evidence revealed the elevated miR-155 levels both in serum and urine of nephrolithiasis patients. The aim of our research was to explore the role of miR-155 in CaOx-induced apoptosis in HK-2 cells. The expression levels of miR-155 in serum and renal tissues were quantified in 20 patients with nephrolithiasis using qRT-PCR assay. ELISA was performed to determine urinary levels of interleukin (IL)-1ß, IL-6 and tumor necrosis factor-alpha (TNF-α). Renal tubular cell model of CaOx nephrolithiasis was established to investigate the role and molelular mechanism of miR-155. Cell viability and apoptosis were assessed by MTT and flow cytometry, respectively. Immunofluoresent staining of LC3 autophagosome and western blotting were performed to evaluate the autophagic activity. Luciferase reporter assay was employed to verify the interaction between miR-155 and PI3KCA/Rheb. PI3K/Akt/mTOR signaling was further examined by western blotting. Serum and renal levels of miR-155 and inflammatory factors were significantly elevated in nephrolithiasis patients than in controls. CaOx treatment caused up-regulation of miR-155 and induced autophagy in renal tubular epithelial cells, while silencing miR-155 or inhibition of autophagy by 3-metheladenine (3-MA) ameliorated CaOx crystal-induced cell injury. PI3KCA and Rheb was identified as downstream targets of miR-155. Moreover, miR-155 activates autophagy and promotes cell injury through repressing PI3K/Akt/mTOR signaling pathway. Taken together, these findings demonstrated that miR-155 facilitates CaOx crystal-induced renal tubular epithelial cell injury via PI3K/Akt/mTOR-mediated autophagy, providing therapeutic targets for ameliorating cellular damage by CaOx crystals.

Inhibition of miR-155 Ameliorates Acute Kidney Injury by Apoptosis Involving the Regulation on TCF4/Wnt/ß-Catenin Pathway.

BACKGROUND: Acute kidney injury (AKI) is a complex clinical disorder with sudden decay in renal function. Ischemia-reperfusion injury (IRI) has been regarded as the main etiology for the occurrence of AKI. MicroRNAs have been consistently shown to be involved AKI. OBJECTIVES: We aimed to investigate the role of miR-155 in AKI and its underlying mechanism. METHODS: Ischemia-reperfusion (I/R)-induced AKI rat model and hypoxia-reoxygeneration (H/R)-induced NRK-52E cell model were established. The concentrations of serum creatinine and blood urea nitrogen were measured to evaluate renal function. Hematoxylin and eosin staining and TUNEL assay were performed to assess the severity of kidney injury. Additionally, quantitative real-time-PCR and western blot analysis were subjected to determine the expression of miR-155, TCF4, and apoptosis-related proteins, respectively. Moreover, cell proliferation and apoptosis were evaluated by Cell Counting Kit-8, bromodeoxyuridine, and flow cytometry analyses, respectively. Luciferase reporter assay was used to validate the direct targeting of TCF4 with miR-155. The protein levels of TCF4 and its downstream proteins in cells were measured by western blot. RESULTS: The expression level of miR-155 was upregulated in both I/R-induced AKI rat model and H/R-treated NRK-52E cells. Moreover, overexpression of miR-155 promoted H/R-induced NRK-52E cells apoptosis and suppressed cell proliferation, while inhibition of miR-155 expression exerted opposite effects. Additionally, TCF4 was identified as a target of miR-155, of which expression was downregulated both in vivo and in vitro. Furthermore, the activity of Wnt/ß-catenin signaling pathway was promoted following overexpression of TCF4 in NRK-52E cells, and this effect was attenuated by the increasing miR-155 expression. CONCLUSION: We demonstrated that miR-155 exacerbated AKI involving the targeting and regulation of TCF4/Wnt/ß-catenin signaling pathway, indicating a novel regulatory network and elucidating a potential target for IRI-induced AKI treatment.

Effect of Baricity of Bupivacaine on Median Effective Doses for Motor Block.

BACKGROUND The median effective dose (ED50) of a drug gives the amount or dose of drug needed to produce effective therapeutic response or desired effect in at least 50% of the population taking it. Our study focused on determining the ED50 required for effective motor block using hyperbaric and plain bupivacaine, and evaluated the influence of baricity on the ED50 required for motor block. MATERIAL AND METHODS A total of 38 patients were randomly assigned into 2 groups according to the baricity of bupivacaine: group P received plain bupivacaine and group H received hyperbaric bupivacaine. The patients were administered 0.5% plain or hyperbaric bupivacaine intrathecally. The dosage of anesthetics in each patient was calculated according to the standard up-down sequential allocation method of Dixon. The first patient in each group received a dose of 7.5 mg bupivacaine, and a dose of 1.0 mg was used as the testing interval. The dose was increased or decreased by 1.0 mg for each patient according to the estimated score of motor block. RESULTS The ED50 required for effective motor block in spinal anesthesia was 7.20 and 10.05 mg in groups H and P, respectively. Their relative motor blocking potency ratio was found to be 0.72. CONCLUSIONS The ED50 for motor block was significantly decreased using hyperbaric bupivacaine intrathecally compared with plain bupivacaine, and the baricity of bupivacaine obviously affected the ED50 for the motor block.

Factors associated with significant liver necroinflammation in chronic hepatitis B patients with cirrhosis.

We determined the association between various clinical parameters and significant liver necroinflammation in patients with chronic hepatitis B (CHB) related cirrhosis. Two hundred patients with CHB related cirrhosis were recruited in the final analysis. Clinical laboratory values and characteristics were obtained from the medical record. We performed analyses of the relationships between independent variables and significant liver necroinflammation by using binary logistic regression analysis and discriminant analysis. Significant liver necroinflammation (grade≥2) was found in 58.0% (80/138) of antiviral therapy patients and 48.4% (30/62) of non antiviral therapy patients respectively. Also, there were some significant differences in serum hepatitis B surface antigen (HBsAg), serum hepatitis B e antigen (HBeAg) and serum hepatitis B virus (HBV) DNA between antiviral therapy and non antiviral therapy patients. After that, aspartate aminotransferase (AST), total bilirubin (TBIL), total bile acid (TBA), prothrombin time (PT), aspartate aminotransferase to platelet ratio index (APRI) and serum HBV DNA were confirmed as independent predictors of significant liver necroinflammation in CHB patients with cirrhosis by univariate analysis and multivariate analysis (p = 0.002, 0.044, 0.001, 0.014, 0.01 and 0.02 respectively). Finally, receiver operating characteristic (ROC) curve analysis and discriminant analysis validated that these six variables together have strong predictive power to evaluate significant liver necroinflammation.

The characteristics and clinical outcome of drug-induced liver injury in a Chinese hospital: A retrospective cohort study.

The aim of this cohort study was to determine the characteristics and clinical outcome of 287 patients with drug-induced liver injury (DILI) in a Chinese hospital.Between January 2008 and January 2013, individuals who were diagnosed with DILI were selected. The complete medical records of each case were reviewed, and factors for the outcome of patients with DILI were extracted and analyzed using univariate and multivariate analysis.Two hundred eighty-seven cases identified as DILI were included in the study. A total of 105 different drugs were considered to be related to the hepatotoxicity. The main causative group of drugs was Chinese herb (n = 111). Liver failure developed in 9 (3.1%) patients, and 2 died (0.7%). Overall, complete recovery occurred in 92 (32.1%) patients. Univariate analysis and binary logistic regression analysis identified the digestive symptoms, jaundice, total bilirubin (TBIL), and direct bilirubin (DBIL) as independent factors for the non-recovery of DILI. Then the prediction model, including digestive symptoms, jaundice, TBIL, and DBIL, was built by using binary logistic regression analysis again. Receiver operating characteristic curve validated the strong power (area under the curve (AUC) = 0.907) of prediction model for predicting the DILI non-recovery.DILI is an important cause of liver test abnormalities, and Chinese herb represented the most common drug group. The factors such as digestive symptoms, jaundice, TBIL, and DBIL have effect on DILI outcomes. The prediction model, including digestive symptoms, jaundice, TBIL, and DBIL, established in this study is really an excellent predictive tool for non-recovery of DILI patients.

A practical community-based response strategy to interrupt Ebola transmission in sierra Leone, 2014-2015.

BACKGROUND: The Ebola virus disease spread rapidly in West Africa in 2014, leading to the loss of thousands of lives. Community engagement was one of the key strategies to interrupt Ebola transmission, and practical community level measures needed to be explored in the field and tailored to the specific context of communities. METHODS: First, community-level education on Ebola virus disease (EVD) prevention was launched for the community's social mobilizers in six districts in Sierra Leone beginning in November 2014. Then, from January to May of 2015, in three pilot communities, local trained community members were organized to engage in implementation of EVD prevention and transmission interruption measures, by involving them in alert case report, contact tracing, and social mobilization. The epidemiological indicators of transmission interruption in three study communities were evaluated. RESULTS: A total of 6 016 community social mobilizers from 185 wards were trained by holding 279 workshops in the six districts, and EVD message reached an estimated 631 680 residents. In three pilot communities, 72 EVD alert cases were reported, with 70.8 % of them detected by trained local community members, and 14 EVD cases were finally identified. Contact tracing detected 64.3 % of EVD cases. The median duration of community infectivity for the cases was 1 day. The secondary attack rate was 4.2 %, and no third generation of infection was triggered. No health worker was infected, and no unsafe burial and noncompliance to EVD control measures were recorded. The community-based measures were modeled to reduce 77 EVD cases, and the EVD-free goal was achieved four months earlier in study communities than whole country of Sierra Leone. CONCLUSIONS: The community-based strategy of social mobilization and community engagement was effective in case detection and reducing the extent of Ebola transmission in a country with weak health system. The successfully practical experience to reduce the risk of Ebola transmission in the community with poor resources would potentially be helpful for the global community to fight against the EVD and the other diseases in the future.

Rapid assessment of knowledge, attitudes, practices, and risk perception related to the prevention and control of Ebola virus disease in three communities of Sierra Leone.

BACKGROUND: The recent outbreak of the Ebola virus disease (EVD) in Sierra Leone has been characterized by the World Health Organization as one of the most challenging EVD outbreaks to date. The first confirmed case in Sierra Leone was a young woman who was admitted to a government hospital in Kenema following a miscarriage on 24 May 2014. On 5 January 2015, intensified training for an EVD response project was initiated at the medical university of Sierra Leone in Jui. To understand the knowledge, attitudes, practices, and perceived risk of EVD among the public, especially after this training, a rapid assessment was conducted from 10 to 16 March 2015. METHODS: Interviews were conducted with 466 participants based on questionnaires that were distributed from 10 to 16 March 2015 by cluster sampling in three adjacent communities, namely Jui, Grafton, and Kossoh Town, in the Western Area Rural District of Sierra Leone. RESULTS: It was found that knowledge about EVD was comprehensive and high. Positive attitude towards prevention was found to be satisfactory. Nearly all participants knew the reporting phone number 117 and had reported some change in behavior since learning about Ebola. More than half (62 %) of the participants had a history of travelling to urban areas, which increases the risk of infection. The multivariable logistic regression analysis showed that community and occupation were variables associated with perceived risk of EVD. CONCLUSIONS: Our study showed that community level social mobilization and community engagement were an effective strategy in the special context.

The combination of three molecular markers can be a valuable predictive tool for the prognosis of hepatocellular carcinoma patients.

Based on molecular profiling, several prognostic markers for HCC are also used in clinic, but only a few genes have been identified as useful. We collected 72 post-operative liver cancer tissue samples. Genes expression were tested by RT-PCR. Multilayer perceptron and discriminant analysis were built, and their ability to predict the prognosis of HCC patients were tested. Receiver operating characteristic (ROC) analysis was performed and multivariate analysis with Cox's Proportional Hazard Model was used for confirming the markers'predictive efficiency for HCC patients'survival. A simple risk scoring system devised for further predicting the prognosis of liver tumor patients. Multilayer perceptron and discriminant analysis showed a very strong predictive value in evaluating liver cancer patients'prognosis. Cox multivariate regression analysis demonstrated that DUOX1, GLS2, FBP1 and age were independent risk factors for the prognosis of HCC patients after surgery. Finally, the risk scoring system revealed that patients whose total score >1 and >3 are more likely to relapse and die than patients whose total score ≤1 and ≤3. The three genes model proposed proved to be highly predictive of the HCC patients' prognosis. Implementation of risk scoring system in clinical practice can help in evaluating survival of HCC patients after operation.

Determination of the median effective dose (ED50) of spinal plain ropivacaine for motor block in adults.

BACKGROUND: The median effective dose (ED50) for motor blockade of spinal plain bupivacaine in adults was previously reported; however, that of ropivacaine is unknown. This study aimed to determine the ED50 for motor blockade of spinal plain ropivacaine in adults, evaluating the effect of age on this parameter. METHODS: 133 patients in need of combined spinal and epidural anesthesia were enrolled in this sequential allocation study. They were divided into 6 groups according to age: 20-30, 31-40, 41-50, 51-60, 61-70, and 71-80 years. Using the up-and-down method of Dixon, the ropivacaine dose was varied according to the preceding patient's reaction. The modified Bromage and hip motor function score was used to evaluate the degree of motor block after administration of ropivacaine. ED50 values were estimated by the method of Dixon and Massey. Other indexes, including the onset of motor block, the analgesia level, and the duration of motor block were also assessed. RESULTS: the ED50 for motor block of intrathecal ropivacaine was 20.96 mg (95 % CI: 19.83-22.16 mg) in 20-30, 19.05 mg (95 % CI: 18.43-19.70 mg) in 31-40, 17.91 mg (95 % CI: 17.10-18.76 mg) in 41-50, 17.91 mg (95 % CI: 16.49-19.44 mg) in 51-60, 16.11 mg (95 % CI: 14.50-17.90 mg) in 61-70, and 15.75 mg (95 % CI: 13.98-17.73 mg) in 71-80 year-old patients. Maximum cephalic analgesic effects were obtained at the L4-T6 and L4-T2 levels, at 5 and 10 min, respectively, after intrathecal administration of ropivacaine in all groups. CONCLUSION: the ED50 for motor block in spinal plain ropivacaine decreases with advancing age, indicating that age has an influence on the potency of spinal ropivacaine.

Effect of Concentration on Median Effective Dose (ED50) for Motor Block of Intrathecal Plain Bupivacaine in Elderly Patients.

BACKGROUND: The aim of the study was to determine the median effective dose (ED50) for motor block of various concentrations of intrathecally administered plain bupivacaine. MATERIAL AND METHODS: Between 2011 and 2013, 64 patients aged ≥70 years, undergoing transurethral, or lower limb surgery with combined spinal and epidural anesthesia in a single hospital were enrolled. The patients were randomized into 3 groups to receive intrathecal 0.75% bupivacaine (Group 1), 0.375% bupivacaine (Group 2) or 0.25% bupivacaine (Group 3). Spinal anesthesia was achieved using injections of up-and-down doses of 0.75%, 0.375%, or 0.25% plain bupivacaine. The first patient in each group received 7.5 mg bupivacaine, and the testing interval was set at 0.75 mg. The efficacy of motor block in both legs was determined using a modified Bromage and a hip motor function scale. The ED50 for motor block was estimated according to the Dixon's up-and-down method. RESULTS: The ED50 for motor block of bupivacaine was 6.10 (95% CI 5.58-6.66) mg in Group 1, 6.04 (95% CI 5.82-6.28) mg in Group 2, and 5.43 (95% CI 5.19-5.67) mg in Group 3. There were significant differences in the ED50 for motor block among the groups (P=0.008). CONCLUSIONS: The ED50 doses for motor block with 3 bupivacaine concentrations were significantly different in elderly patients; the ED50 dose of 0.75% bupivacaine being significantly higher than that of 0.25% bupivacaine.

Hepatic sarcoidosis mimicking liver cancer.

We present a case of a 50-year-old woman with multiple occupations in the liver. Liver cancer was strongly suspected initially according to the results of imaging examination. However, sarcoidosis was confirmed subsequently by liver biopsy, so methylprednisolone was then prescribed and the patient showed favorable therapeutic response. This case report suggests that hepatic mass in Chinese patients without any history of hepatitis virus infection should be carefully investigated before giving a diagnosis of liver cancer. The report also reminds us that the clinical presentation of sarcoidosis is complex and involvement of a single extra-pulmonary organ should not be ignored.

Serum microRNA-29 levels correlate with disease progression in patients with chronic hepatitis B virus infection.

OBJECTIVE: To investigate the role of serum microRNA-29 (miR-29) as a biomarker for the prediction of disease progression in patients with chronic hepatitis B virus (HBV) infection. METHODS: Using real-time quantitative polymerase chain reaction assay, serum miR-29a, miR-29b and miR-29c levels were measured in patients with chronic HBV infection, and the correlation between serum miR-29 levels and the participants' liver biochemistry, fibrotic stage and necroinflammation grade were also evaluated. RESULTS: Altogether 91 patients with chronic HBV infection were divided by fibrotic stage into S0/1 (no or mild fibrosis), S2/3 (progressive fibrosis) and S4 (cirrhosis) subgroups, and 12 healthy individuals were also included in the study. Serum miR-29a and miR-29c in S0-3 were significantly higher than those in S4 patients (P < 0.001); however, the difference between S0/1 and S2/3 patients was not significant. miR-29b levels were higher in S0/1 patients than in other patient groups, but did not differ between S2/3 and S4 patients. At fibrotic stages of S0/1 and S2/3, patients with no or mild liver inflammation (G0/1) tended to express higher miR-29 levels than those with advanced inflammation (G2-4) (P > 0.05). miR-29a-c showed significant correlation with alanine transaminase levels (P < 0.05 for miR-29a, miR-29b and miR-29c) in S0-3 patients. The expression of miR-29 was highest in immune-tolerant patients (P < 0.001). CONCLUSIONS: Serum miR-29 levels are negatively correlated with liver fibrotic stages and necroinflammation grades in patients with chronic HBV infection. miR-29 appears to be a novel biomarkers for predicting disease progression in these patients.

Determination of the median effective dose for motor block of intrathecally administered different concentrations of bupivacaine in younger patients. The influence of solution concentration.

OBJECTIVE: To determine the median effective dose (ED50) for motor block of intrathecally administered different concentrations of bupivacaine in younger patients. METHODS: This study was conducted at the Department of Anesthesiology, The First College of Clinical Medical Science, China Three Gorges University, China, on 40 American Society of Anaesthesiologists (ASA) I-II younger patients (18-40 years) undergoing lower limb surgery. These patients were classified into 2 groups according to a computer-generated random number table (Group A - 0.375% bupivacaine, and Group B - 0.75% bupivacaine). Spinal anesthesia was established by bolus administration of various up-and-down doses of 0.375%, or 0.75% plain bupivacaine. These up-and-down doses were determined by Dixon's up-and-down method. The first patient of each group was given 7.5 mg bupivacaine, and the testing interval was set at 0.75 mg. The dose was up, or down, 0.75 mg increments according to the failure or success of the preceding patient's motor block. The degree of motor block after intrathecal administration of bupivacaine was evaluated by the modified Bromage and Hip motor function score. The ED50 were estimated from the up-down sequences using the method of Dixon and Massey. RESULTS: Our results showed that ED50 for motor block of intrathecal bupivacaine was 8.890 mg in Group A, and 9.998 mg in Group B, and the relative motor blocking potency ratio was 1.12. CONCLUSION: The ED50 for motor block of intrathecally administered bupivacaine with higher concentration was slightly higher than that of a lower concentration.

Fever of unknown origin: report of 107 cases in a university hospital.

Fever of unknown origin (FUO) is a challenging problem in clinical practice. Evaluation of patient's characteristics may illustrate the etiologies of FUO. In present study, 107 patients with FUO hospitalized in our inpatient department between 2010 and 2011 were investigated. The median age of the patients was 48 years (15-94). The median fever duration was 8.5 weeks (3-104). The median hospital stay was 8.5 days (1-51). Etiologies of FUO were identified as follows: infectious diseases 32 (29.9%), malignancies 19 (17.8%), inflammatory rheumatic diseases 18 (16.8%), and miscellaneous diseases 15 (14.0%). In 23 (21.5%) patients, the diagnosis remained unclear. Infection group had relative shorter average fever duration and hospital stay than other groups. Shortened mean fever duration was observed in geriatric age group. In conclusion, as the most common cause of FUO in the present study, infectious cases had relative shorter average fever duration and hospital stay, and geriatric patients had shortened average fever duration as well.

[Study on type I interferon and phospho-IRF3 in murine liver dendritic cells after intervened by HBV].

OBJECTIVE: To detect the secretions of type I interferon and the expressions of phospho-IRF3 in murine liver dendritic cells intervened by HBV. METHODS: 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. RESULTS: 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. CONCLUSIONS: The type I interferon secretion and the phospho-IRF3 expression were decreased in murine liver dendritic cells when intervened by HBV.

Correlation of the expression of toll-like receptors in monocyte-derived dendritic cells with prognosis of chronic severe hepatitis B.

OBJECTIVE: This study aims to measure the expression of toll-like receptors (TLR) in monocyte-derived dendritic cells (MoDC) from chronic severe hepatitis B (CSHB), to assess the contribution of TLRs in CSHB. METHODS: Peripheral blood was collected from 40 CSHB patients, 30 chronic hepatitis B (CHB) patients, and 30 healthy individuals who served as healthy controls (HCs). Purified monocytes were isolated by a combination of Histopaque-1.077 and CD14 Microbeads. MoDCs were induced with granulocyte macrophage colony-stimulating factor and interleukin-4 for 6 days from CD14(+) monocytes. The expression of TLRs in MoDC was measured using real-time PCR and flow cytometry. RESULTS: The expressions of TLR-1, -2, -7 were significantly higher in MoDC of CSHB than that of HCs, of which the level of TLR-3 was decreased. Particularly in CSHB patients, the TLR-3 expression was further decreased compared to CHB patients. In non-survival CSHB patients, TLR-3 level was significantly decreased, while TLR-2 expression was dramatically increased. Linear correlation analysis demonstrated significant correlations between TLR-3 level and disease severity markers (total bilirubin, prothrombin activity, creatinine, white blood cell count, and maximum volume of ascitic fluid) in individual CSHB patients. CONCLUSIONS: TLR-2 and TLR-3 may be involved in the pathogenesis of CSHB, and TLR-3 may influence the prognosis of CSHB.

[Expression of nuclear factor-κB and its regulated cytokines in dendritic cells in patients with chronic severe hepatitis B and its clinical significance].

OBJECTIVE: 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). METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.

[Effect of fresh gas flow on isoflurane pharmacokinetics during anesthesia induction].

OBJECTIVE: To investigate the effect of different fresh gas flow (FGF) rates on isoflurane pharmacokinetics during anesthesia induction. METHODS: 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. RESULTS: 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%. CONCLUSIONS: 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.