Multiple Mild Stimulations Reduce Membrane Distribution of CX3CR1 Promoted by Annexin a1 in Microglia to Attenuate Excessive Dendritic Spine Pruning and Cognitive Deficits Caused by a Transient Ischemic Attack in Mice / 神经科学通报·英文版

A transient ischemic attack (TIA) can cause reversible and delayed impairment of cognition, but the specific mechanisms are still unclear. Annexin a1 (ANXA1) is a phospholipid-binding protein. Here, we confirmed that cognition and hippocampal synapses were impaired in TIA-treated mice, and this could be rescued by multiple mild stimulations (MMS). TIA promoted the interaction of ANXA1 and CX3CR1, increased the membrane distribution of CX3CR1 in microglia, and thus enhanced the CX3CR1 and CX3CL1 interaction. These phenomena induced by TIA could be reversed by MMS. Meanwhile, the CX3CR1 membrane distribution and CX3CR1-CX3CL1 interaction were upregulated in primary cultured microglia overexpressing ANXA1, and the spine density was significantly reduced in co-cultured microglia overexpressing ANXA1 and neurons. Moreover, ANXA1 overexpression in microglia abolished the protection of MMS after TIA. Collectively, our study provides a potential strategy for treating the delayed synaptic injury caused by TIA.

Establishment of a candidate reference method for serum glycated albumin based on liquid chromatography tandem mass spectrometry / 中华检验医学杂志

Objective:To establish an accurate quantitative method for the determination of serum glycated albumin (GA) based on isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS).Methods:This study optimized the ID-LC/MS method recommended by the Japan Society of Clinical Chemistry (JSCC). Albumin (Alb) was purified by anion exchange chromatography. The standard solution of purified sample, lysine (Lys) and deoxyfructosyl lysine (DOF-Lys) and their mixed internal standard were weighed accurately by weight method. The Alb was hydrolyzed to amino acid, and Lys and DOF-Lys were detected to determine GA. The method was evaluated and compared with the routine method.Results:The purity of Alb was 100%. The R 2 ranges of Lys and DOF-Lys linear standard curves were 0.997 8-0.999 9 and 0.999 4-1.000 0. The results of three-day precision evaluation of low, medium and high concentrations of serum were 0.69%-1.97%, 1.13%-2.33% and 1.37%-2.54% in intra, inter and total Coefficient of variation ( CV). In the accuracy evaluation, the deviations between the results of three concentration standards and the certified value were -2.56%, -1.87% and-2.94%. The matrix effects of Lys and DOF-Lys were 89.92% and 109.98%, and the carryover rates were -1.13% and -3.27% respectively. The detection limit and quantitative limit of Lys were 0.075 nmol/g and 0.248 nmol/g, and the DOF-Lys were 0.007 nmol/g and 0.024 nmol/g. The relative expanded uncertainty was 1.60%-2.03%. The fitting regression curve R 2 with the routine method was 0.970 7. Conclusions:A method was established for the detection of serum GA. The method evaluation was accurate and reliable. It was expected to be a candidate reference method for the detection of serum GA.

Daclatasvir-sofosbuvir combination therapy for chronic hepatitis C virus infection: progress in clinical studies / 中华肝脏病杂志

All-oral, pan-genotypic combination of direct-acting antiviral agents is currently clinically prescribed trends in the management of chronic hepatitis C virus infection. The combination of daclatasvir and sofosbuvir has proven strong antiviral activity across all common genotypes in clinical trials and real world studies. Furthermore, it can be safely used in patients with advanced liver disease, HCV/HIV co-infection, or HCV recurrence after liver transplantation, more patients will gain clinical benefits. We aim to summarize the clinical progress about this combination.

The application of IFCC reference methods in methodological evaluation of glycated hemoglobin measurement systems / 中华检验医学杂志

Objective To perform a methodological evaluation study of 4 HPLC based systems and a capillary electrophoresis based system , with the International Federation of Clinical Chemistry and Laboratory Medicine ( IFCC) glycated hemoglobin reference method as a comparative method .Methods 40 hemolysis samples of variety concentrations were prepared .The samples were measured by IFCC reference method and 5 glycated hemoglobin testing systems , respectively and trueness verification was performed according to the Clinical &Laboratory Standards Institute (CLSI) guideline EP9-A3.Whole blood samples were used to test the systems'precision, linearity and analytical interferences .Results The average CV of IFCC reference method results of 40 hemolysis samples was 1.4％( range from 0.2％to 2.5％).No outlier was found in the results of the 5 testing system.The slopes ranged from 0.9902 to 1.0267, and intercepts from -0.1526 mmol/mol to +0.1512 mmol/mol, squared correlation coefficient from 0.9962-0.9971, biases at two medical decision level were less than 0.3％HbA1c.Within-laboratory precisions were less than 2％ (NGSP unit).Bias between all test results and predicted results were less than 5％.High concentration of glucose showed certain interference to glycated Hemoglobin tests but had little influence in clinical practice.Conclusions The results of the 5 testing system are comparable to the IFCC reference method , the results of precision and linearity evaluation meet the requirements of related guidelines .

Strategy for virologic response-guided therapy in integrated diagnosis and treatment of hepatitis C in the era of precision medicine in China / 临床肝胆病杂志

At present, long-acting interferon combined with ribavirin is still a major therapeutic method for hepatitis C in China, and precision medicine can help cure more patients with hepatitis C. This article briefly introduces the diagnosis and treatment of hepatitis in China and the importance of the strategy of virologic response-guided therapy (RGT), and analyzes the influencing factors for the achievement of sustained virologic response. HCV RNA quantitation is the foundation of RGT strategy, and HCV RNA quantitation with a high specificity performed at crucial time points during treatment helps to evaluate response accurately, realize the individualized treatment of hepatitis C, and improve the cure rate of hepatitis C.

The expression of circulating microRNA-595 in patients with hepatitis B virus-related liver failure and its relationship with disease prognosis / 中华传染病杂志

Objective To measure the expression of circulating microRNA (miRNA)in patients with hepatitis B virus (HBV)-related liver failure and its relationship with disease prognosis.Methods The miRNA expressions in serum of 5 patients with HBV-related liver failure and 5 healthy control subjects were compared using Exiqon miRCURY LNATM miRNA microarray.The sera from 20 patients with chronic hepatitis B (CHB),20 patients hepatitis B related cirrhosis,50 patients with HBV-related liver failure and 40 healthy persons in Ruijin Hospital were collected.The relative expression of miRNA-595 was measured using quantitative real-time polymerase chain reaction (PCR).The relative expressions of miRNAs among groups were analyzed using student t test,the correlations were analyzed by Pearson and Spearman correlation.Results Microarray informed that 92 miRNAs changed significantly in patients with HBV-related liver failure,and miRNA-595 increased most significantly.The results of real-time PCR showed that the relative expressions of miRNA-595 ,miRNA-300 and miRNA-122 were 6.03 (t=3.134, P =0.003),3.12 (t=7.221 ,P <0.01)and 2.77 (t=2.671 ,P =0.021),which were higher compared to those in healthy control group.In the analysis of the relationship between miRNA-595 expression and disease prognosis in patients with HBV-related liver failure,the relative expressions of miRNA-595 in patients with CHB,hepatitis B related cirrhosis and HBV-related liver failure were 2.26 (t =3.780,P =0.001),3.32 (t = 6.111 ,P < 0.01)and 6.03 (t = 3.134,P = 0.003),respectively,which were all increased compared to that of the healthy control.The relative expression of miRNA-595 of patients with HBV-related liver failure was 2.66 times (t=2.450,P =0.043)higher than that of patients with CHB. When dividing patients according to prothrombin activity,miRNA-595 increased significantly in patients with early stage liver failure.When dividing patients according to model of end-stage liver disease (MELD) score,MELD score was positive correlated with the expression of miRNA-595 when MELD score was under 30 (r=0.673,P =0.004).The expression of serum miRNA-595 in survival group (11 .08,n=23) was higher than that in non-survival group (3.67,n = 27,t =4.309,P =0.041).Conclusions The expressions of miRNA595 ,miRNA-300 and miRNA-122 are all increased in patients with HBV-related liver failure,especially the expression of circulating miRNA-595 at early stage of the disease.The miRNA-595 may be used as a new serum biomarker for monitoring the severity of disease.

Analysis on the allergens in allergic rhinitis patients in Changzhou district of Jiangsu / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the distribution of positive allergens in patients with allergic rhinitis in Changzhou.@*METHOD@#A total of 4242 cases with allergic symptoms and signs underwent skin prick test with 16 kinds of standardized allergens. The common allergens were found out. The samples were stratified on seven age groups to analysis the distributive characteristics of allergens responsible for each group. The samples were also stratified on children group and adults group to analysis their response degree to dust mite.@*RESULT@#2638 cases (62.19%) had the positive reaction, including male 1380 (52.3%) cases, female 1258 (47.7%) cases. Dermatophagoides farinae and dermatophagoides pteronyssinus had the highest positive ratio (85.86%, 83.55%) in inhalation group, followed cockroach (11.49%) and Humulus L. (9.82%), fungus (2.92%) in the lower positive ratio. Shrimp (5.76%) was the most common allergen in food group. The distribution of common allergens in different age groups was variable. There was significantly higher sensitivity to dust mite between children group compared to adults group (P < 0.01).@*CONCLUSION@#The study shows that Dust mite was the mostly responsible common allergen in Changzhou district. The distribution of common allergens in children group and adults group was variable.

Liver histological changes and influencing factors in patients with chronic hepatitis C / 中华传染病杂志

ObjectiveTo investigate the histological features as well as the factors influencing liver disease progression in Chinese patients with chronic hepatitis C (CHC). MethodsA total of 102 CHC patients who underwent percutaneous liver biopsy between August 2007 and May 2010 were recruited. Age, gender, body mass index (BMI) and transmission route of recruited patients were recorded. Serum levels of alanine transaminase (ALT) and aspartate transaminase (AST), HCV genotypes, HCV viral load and liver histological changes were detected. Statistical analysis was done by t test and Logistic regression. ResultsThe serum levels of ALT and AST in CHC patients with histological activity index (HAI) ≥4 were much higher, while platelet (PLT) counts were lower than those with HAI ＜4(t=2.209, 2. 298 and 2. 565, respectively; all P＜0.05). Likewise, in patients with F≥3, the serum levels of ALT and AST as well as the mean age and the duration of infection were significantly elevated compared with F ＜ 3 group ( t = 3.497, 2. 758, 2. 340 and 2. 570,respectively; all P＜0. 05), while PLT counts were much lower (t = 2. 761, P=0. 007). The unvariate predictors for HAI≥4 were female, ALT＞1 × upper limits of normal (ULN), AST level,F≥3, HCV RNA≥6 lgIU/mL and PLT counts. By mutivariate analysis, the Ishak stage score was the only independent predictor for HAI≥4 (OR 3.098, 95％CI 1.884-5. 092; P＜0.01). Finally,the univariate predictors for F≥3 were age, BMI≥24 kg/m2 , ALT＞1 × ULN, AST level, HAI≥4,PLT counts and duration of infection≥ 15 years. Multivariate analysis revealed that age (OR 1. 074,95％CI 1.006-1. 146; P=0.033), ALT level (OR 1. 035, 95％CI 1.015－1.055; P＜0.01), ASTlevel (OR 0. 969, 95％CI 0. 948－0. 990; P=0. 005), the duration of infection ≥15 years (OR 37. 215, 95％CI 5. 816－238. 127; P＜0.01) and HAI≥4 (OR 1. 939, 95％CI 1. 426－2. 636; P＜0.01) were independent predictors for F≥ 3. ConclusionAge, ALT level, AST level, duration of infection≥15 years, HAI≥4 are independent predictors for liver fibrosis.

Epidemiological and clinical characteristics of 323 chronic hepatitis C patients / 中华传染病杂志

Objective To analyze epidemiological and clinical characteristics of chronic hepatitis C (CHC) patients. Methods The clinical data of 323 CHC patients were collected. The transmission modes, clinical manifestations and virological features were recorded. The liver histological change was also analyzed in 39 cases whose liver biopsy samples were available. The comparison between two groups was performed by t test. Results Among the 323 CHC patients, 135 cases (41. 8%) had history of blood or blood products transfusion. Sixty-seven cases (20. 7%) had undergone surgery and trauma operation. Nineteen cases (5. 9%) had history of Chinese medicine acupuncture. Eighteen cases (5.6%) had undergone hemodialysis. Two patients (0.6%) were infected through vertical transmission. Twenty-one cases (6. 5%) had history of intravenous drug use and two cases (0. 6%) had history of unsafe sexual contact. The possible transmission routes for the other 78 cases (24.1%)were unknown. Fourteen patients (4. 3%) were co-infected with hepatitis B virus (HBV). The major prevalent genotypes were hepatitis C virus ( HCV) genotype lb and 2a, which were 145 cases (65. 3%) and 21 cases (9. 5%) respectively. HCV viral loads were as high as 1 × 105 IU/mL in 74 cases (26. 1%) and 1× 106 IU/mL in 103 cases (36. 4%). Twenty-three patients (7.1%) developed obvious clinical manifestations. Among 39 patients undergoing liver biopsy, 14 cases (35. 9%) had hepatic inflammation activity index (HAI)≥4, six cases (15. 4%) had fibrosis stage (F) ≥3, four cases (10. 3%) had HAI≥4 and F≥3. Conclusions The most common HCV transmission modes are blood transfusion and use of blood products. However, surgery and trauma operation should be paid more attention. Besides blood transfusion, the transmission modes of intravenous drug injection, hemodialysis and traditional Chinese medicine acupuncture are increasing. The major HCV genotypes are lb and 2a. The viral loads of most patients are relatively high. Most patients infected with HCV don't show any obvious hepatitis symptoms and physical signs. However, the liver biopsy results from 39 patients suggest that most patients develop liver histological changes.

The expression of microRNAs in the peripheral blood mononuclear cells of the patients with hepatitis B virus infection / 中华传染病杂志

Objective To investigate the expressions of microRNAs (miRNA) and cytokines in the peripheral blood mononuclear cells (PBMCs) of patients with hepatitis B virus (HBV) infection and analyze their relationship with inflammation. Methods PBMCs were collected from acute hapatitis B (AHB) patients of 3 groups, including acute episode, virus clearance, recover period, and chronic hepatitis B (CHB) of mild, medium, severe type, and HBV-related liver cirrhosis. Each group included 20 patients, and 17 healthy donors were as control. Reverse transcription-polymerase chain reaction was used to measure miRNA146, miRNA155, miRNA181, interferon (IFN)-α, IFN-β,interferon induced gene 54 (ISG54) and interferon regulate factor 5 (IRF5). Comparisons among groups were done by one factor analysis of variance. Results The expression of miRNA155 was high in acute episode of AHB (2. 386± 1. 835), and higher than healthy control (1. 498± 1. 276) (F=1. 137,P-=0. 045), while reduced in acute episode, virus clearance (1. 633±2. 291), and recover period (0. 642±0. 836) (F=2. 122,P=0. 022). The expressions of IFN-α and IFN-β in AHB reduced in acute episode, virus clearance and recover period ( F = 1. 880, P = 0. 038 ; F= 1. 835, P = 0. 048).The expression of miRNA155 in AHB is closely correlated with IFN-α and IFN-β (r = 0. 483, P=0. 004; r= 0. 660, P= 0. 0002, respectively). In addition, in HBV infectious patients, the expression of miRNA155 was correlated with alanine transarninase (ALT), serum bilirubin (TBil) (r=0. 342,P=0. 006; r=0. 322, P= 0. 011, respectively), but not with HBV DNA load. Compared with healthy control (1. 307+ 0. 935), miRNA181 was higher in patients with HBV infection (F= 2. 072, P=0. 045) except AHB in recover period (1. 873±0. 998). There was no statistical difference in the miRNA146 expression between various groups. Conclusions The exprossion of miRNAs might be involved in the host anti-HBV immune response during HBV infection, and closely correlated with expression of IFN-related immune factors.

The expression and function of retinoic acid-inducible gene Ⅰ in monocyte-derived dendritic cells in patients with hepatitis B virus infection / 中华传染病杂志

Objective To investigate the expression and function of retinoic acid-inducible gene Ⅰ(RIG-Ⅰ) in monocyte-derived dendritic cells (MoDC) at different stages of hepatitis B virus(HBV)infection and to explore the role of RIG-Ⅰ in the disease progression after HBV infection. Methods Peripheral blood samples were collected from 28 hepatitis B virus-infected persons, including 21 cases of chronic hepatitis B (CHB) and 7 of acute hepatitis B (AHB). Eighteen healthy subjects were recruited as controls. Purified CD14~+ monocytes were isolated by CD14 microbeads. MoDCs were induced from CD14~+ monocytes with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4 for 7 days, and then were infected with vesicular stomatitis virus (VSV) to stimulate RIG-Ⅰ expression. The mRNA expression levels of RIG-Ⅰ, interferon (IFN )-promoter stimulating factor-1 (IPS-1) and IFN-β at 16 hours and 24 hours after infection with VSV were measured by real-time quantitative polymerase chain reaction (PCR). Data with normal distribution were tested by analysis of variance. Continuous variables between groups were compared using Mann-Whitney U test. Comparison among multiple groups was done by Kruskal-Wallis test. Results The expression levels of RIG-Ⅰ in MoDCs from CHB patients were significantly lower than those in AHB patients and healthy controls at 16 hours (2.44±2.03, 19. 54±3. 15, 21. 48±8. 39, respectively; F=7.451,P=0.002) and 24 hours (2. 68±2. 93, 10. 31 ±3. 88, 14. 01 ±5. 04, respectively, F = 7. 908, P = 0. 001)following VSV stimulation. The IPS-1 levels in both CHB patients and AHB patients were higher than those in healthy controls at 16 hours (2. 05±l. 08, 1. 99±1. 56, 0. 60±0. 31, respectively) F=7.246,P =0.003) and 24 hours (2. 27±2. 16, 3.24 ± 1.21, 1. 08±0. 73, respectively; F= 13. 598, P = 0. 001).Furthermore, the IFN-β expression levels were significantly lower in CHB patients compared to AHB patients and healthy controls at 16 hours and 24 hours after VSV stimulation. Conclusions The expressions of RIG-Ⅰ and IPS-1 in MoDC are abnormal in HBV infected persons, which indicates that RIG-Ⅰ signaling pathway might be blocked by HBV. The impaired function of MoDC may play a role in HBV infection and chronicity.

The role of liver-targeted CD4+ CD25+ Foxp3+ regulatory T cells in the patients with chronic hepatitis B / 中华传染病杂志

Objective To study the correlation between the percentage of intrahepatic regulatory T cell (Treg) and liver inflammatory activity in chronic hepatitis B (CHB) patients.Methods Twenty-six cases of CHB patients admitted to First Department of Internal Medicine,Nagasaki University School of Medicine,Sakamoto,Nagasaki,Japan were enrolled and performed liver biopsy in this study.CD3+cell and Foxp3+ cell in liver were detected by immunohistochemistry.the percentage of Foxp3+/CD3+cell was determined.Clinical data including alanine aminotransferase (ALT).aspartate aminotransferase(AST), hepatitis B virus (HBV) DNA level and the inflammatory activity of histological activity index score (HAD of liver pathology using Knodell evaluating system were collected.The data was analyzed by SPSS 13.0 software.Results Foxp3+Tregs in serial sections of CHB accumulated mainly in the portal area.There Was a significance correlation between the percentage of Foxp3+/CD3+ cell and liver parenchyma inflammation (P=0.007 6).Moreover,Foxp3+Treg in CHB patients with high serum ALT or AsT level presented in a higher frequency than in patients with low AIT or AST level.The difference between these tWO groups was statistically significant (rALT=O.438,PALT=0.025; fAST=O.436,Past=O.026).There Was a tendency between the percentage of Foxp3+/CD3+ cell in liver and HBV DNA level.however the correlation was not statistically significant.Conclusion It is suggested that CD4+ CD25+ Foxp3+Treg may play a major role in the pathogenesis of liver injury in the CHB patients.

Histological findings and its influencing factors of HBeAg-negative chronic hepatitis B virus-infected patients with persistently normal alanine aminotransferase levels / 中华传染病杂志

Objective To identify the histological features as well as factors influencing the course of HBeAg-negative chronic hepatitis B virus ( HBV)-infected patients with persistently normal alanine amino-transferase (ALT) levels (PNAL). Methods Ninety-eight HBeAg-negative chronic HBV-infected patients with PNAL who underwent percutaneous liver biopsy were recruited from October 2003 to March 2008. The ALT level, HBV markers, HBV DNA level and liver histological changes were detected. Comparison of means was done by t test and single factor analysis of variance. Nonparametric statistics was done by Marm-Whitey U test and Kruskal-Wallis test. Analysis of independent risk factor was done using Logistic model. The dianostic value of ALT level to significant liver histological changes was evaluated by receiver performance curve. Results Twenty-two point four percent and 17.3% of subjects had the histological activity index (HAI)≥4and fibrosis (F) score≥3 respectively. Subgroup analysis showed that subjects with ALT＞0.50 × upper limit of normal (ULN) had a significantly higher rate of HAI≥4 and F score≥3 than those with ALT≤0.50×ULN (HAI≥4:36.4% vs 11.1%, χ2 =8.881, P=0.003;F score≥3:27.3% vs 9.3%, χ2 =5.487, P= 0.019, respectively), and older subjects (more than 45 years old) had a higher proportion of HAI ≥4 than the younger (33.3% vs 13.4%, χ2 =4.923, P=0.027). Multivariate Logistic regression analysis revealed that a decade increase in age was the independent predictor of HAI≥4 (OR=2.410, P=0.023).Receive operating characteristic (ROC) curve showed that 87.0% and 90.7% of subjects with ALT＜0.50× ULN had histological changes of HAI＜4 and F score＜3 respectively. The proportions of HAI≥4 and F score≥3 in subjects with HBV DNA＜1×104 copy/mL were 14.9% and 12.8%, respectively. Conclusions Significant histological changes may be present in part of the subjects with persistently normal ALT and different HBV DNA levels, so that liver biopsy is very important, especially in those with age ＞45 years.Half time the ULN may serve as an appropriate cutoff value of normal ALT level for managing Chinese HBeAg-negative chronic HBV-int'ected patients.

Determination of Chlorogenic Acid and Baicalin in Xiaoer Qingrening Granula by HPLC / 中国药房

OBJECTIVE:To establish a HPLC method for the determination of chlorogenic acid and baicalin in xiaoer qingrening granula.METHODS:The determination was performed on Symmetry Shield RP C 18 ,the mobile phase of chlorogenic acid consisted of acetonitrile-0.4%phosphoric acid soluton(13∶87)and that of baicalin consisted of methanol-water-glacial acetic acid(50∶50∶1),the detection wavelengths of chlorogenic acid and baicalin were327nm and274nm,respectively,the flow rate was1.0ml/min,sample size was20?l and column temperature was25℃.RESULTS:The respective linear sample size range for chlorogenic acid and baicalin was0.1040?g～1.040?g(r=0.9998)and0.6240?g～6.240?g(r=0.9997),with average recoveries respectively at96.4%(RSD=1.4%)and98.0%(RSD=1.1%).CONCLUSIONS:The method is simple,accurate,reproducible and specific,and it can be used for the quality control of xiaoer qingrening granula.

Effect of homocysteine-mediated endoplasmic reticulum stress on lipid metabolism in hepatocytes / 中国病理生理杂志

AIM:To explore the effect of homocysteine (Hcy)-mediated endoplasmic reticulum stress on lipid metabolism in hepatocytes.METHODS: HepG2 cells used in the study were treated with 5 mmol/L Hcy. The concentrations of Hcy, triglycerides (TGE) and cholesterol (CHO) in the cells were measured. In high methionine diet-induced hyperhomocysteinemia C57BL/6 mice, the concentrations of TGE and CHO in hepatocytes were analyzed. The mRNAs and proteins expressions of glucose-regulated protein 78 (GRP-78) and sterol regulatory element binding protein (SREBP-1) were also assessed.RESULTS: The concentrations of Hcy and lipids (TGE, CHO) in HepG2 cells at different time point were elevated after treated with 5 mmol/L Hcy (P0.05,vs 0 week). The mRNAs and proteins expressions of GRP-78 and SREBP-1 in mice at different time point after high methionine diet were higher than that at 0 week (P