BackgroundGlaucoma is an optic neuropathy caused by structural damage of the optic nerve,and its early diagnosis is critical for arresting the irreversible damage of visual function. Optical coherence tomography (OCT) allows an early diagnosis of glaucoma by the measurements of the optic disc and retinal nerve fiber parameters. Objective This study was carried out to evaluate the effects of optic disc tomography and the measurement of the retinal nerve fiber layer (RNFL)thickness by spectral-domain OCT on the diagnosis of glaucomatous eye. MethodsIt was a noninterventional, cross-sectionalstudy. The optic disctopographic parameters and total and regional RNFL thickness were measured by RTVue OCT in 62 normal eyes and 67 glaucomatous eyes. The area under the receiver operating characteristic curve( ROC ) was used to assess the ability to differentiate glaucoma eyes from normal eyes of each testing parameter. This trial complied with the Helsinki Declaration and was approved by the Clinical Trial Ethic Committee of Beijing TongrenHospital. All of the participants signed the written informed consent before any medical examination. Results In the comparison of demography ,the ages of patients, the mean deficiency( MD ) and pattern standard difference( PSD ) of perimetry were obviously larger in the glaucoma group, primary open angle glaucoma ( POAG ) group and primary closure-angle glaucoma(PACG) group than those of normal controls( P＜0. 01 ). No significant differences were found in the disc area between a total glaucoma group, POAG group or PACG group and normal group ( P =0. 101,0. 741 and 0. 652, respectively) ;however, the average RNFL thickness between normal eyes and glaucomatous eyes were significantly different( 109. 758 μm versus 79. 539 μm, P＜0. 01 ). Among the eight regions around the optic disc, the thickest RNFL located at the inferotemporal( 150. 109 μm) and superotemporal( 146. 105 μm) regions in normal eyes,and at the superotemporal( 104. 354 μm) and inferotemporal( 102. 436 μm) regions in glaucomatous eyes. Both in normal and glaucomatous eyes,the thinnest RNFL located at the nasal(NU+NL) and temporal(TU + TL) regions. For optic disc topographic parameters,the highest ROC were observed in rim volume( ROC--0. 850,0. 841 and 0. 862 in total glaucoma,POAG and PACG, respectively) and vertical cup/disc ratio( ROC =0. 840,0. 849 and 0. 830 in total glaucoma,POAG and PACG,respectively), and the sensitivities for specificity cutoff set at 80％ were 73.1％ and 76. 1％ in total glaucoma,73.0％ and 81.1％ in POAG and 73.3％ and 70.0％ in PACG, respectively. For RNFL thickness ,the highest ROC was observed in average RNFL( ROC =0. 925,0. 910 and 0. 942 in total glaucoma, POAG and PACG,respectively) ,and the sensitivities for specificity cutoff set at 80％ were 89. 6％ ,89.2％ and 90. 0％ in total glaucoma,POAG and PACG, respectively. Among the eight regions around the optic disc, RNFL thickness of region IT achieved the highest ROC, RNFL thickness of region TU and TL had the lowest ROC. Conclusions RTVue OCT appears to be of fair discriminating ability in distinguishing normal from glaucomatous eyes. RTVue OCT shows promise for the diagnosis of glaucoma.

OBJECTIVETo study the effects of hepatitis B virus (HBV) infection on the expression of Furin, an important proprotein convertase, in liver cells to provide insights towards its potential as a therapeutic target for improved antiviral efficacy.

METHODSFurin expression was measured in human liver specimens (infected tissues from patients with chronic HBV hepatitis vs. normal tissues from healthy donors) and in hepatoma cell lines (HBV-infected HepG2.2.15 cells vs. uninfected parental cell lines HepG2) using quantitative real-time RT-PCR (for mRNA), western blotting and immunohistochemistry (for protein).

RESULTSCompared to the uninfected tissues and cells, the HBV-infected tissue and cells showed down-regulated expression of furin at both the mRNA and protein levels. In particular, the HepG2.2.15 cells showed -50% less furin mRNA expression than the HepG2 cells and the difference was statistically significant (P less than 0.05).

CONCLUSIONHBV may suppress the host cell's expression of furin, possibly to benefit its survival and replication in the host cell.

Cell Line ; Furin ; metabolism ; Gene Expression Regulation ; Hep G2 Cells ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; metabolism ; Host-Pathogen Interactions ; Humans ; Liver ; metabolism ; virology ; Proprotein Convertases ; metabolism ; Virus Replication

OBJECTIVETo establish a sequential antiviral regime and evaluate its efficacy in patients with chronic hepatitis B using a controlled trial.

METHODSSeventy-four patients with chronic hepatitis B were divided into 3 groups: 30 cases were enrolled in the sequential antiviral group in which patients received eight-week treatment with thymosin alpha1 (1.6 mg/time, subcutaneous injection, 2 times/week), six-month treatment with interferon (500 MU/ times, muscle inject, every other day) begun in the fifth week of the therapeutic course, and lamivudine treatment (100 mg/days) begun 2 months later after HBeAg seroconversion or just after the withdrawal of interferon to more than eighteen months. Fourteen cases were enrolled in combination group in which patients received six-month treatment with interferon and thymosin alpha1 simultaneously in the same manner as in sequential antiviral group. Thirty cases were enrolled in lamivudine group in which patients received more than eighteen-month treatment with lamivudine.

RESULTSThe temporary rates of HBeAg seroconversion and normalization of alanine aminotransferase (effective rate) in sequential antiviral group, combination group and lamivudine group were 76.7%, 78.6% and 13.3%, respectively. The effective rates of sequential group and combination group were very similar, and significantly higher than that of lamivudine group (P less than 0.01). Long-term efficacy rates were 76.7%, 57.1% and 16.7% among the three groups, respectively. The long-term effective rate of sequential group was relatively higher. The rate of liver damage sensitive period in sequential antiviral group and combination group was 47.7%. The time of onset was from 2 to 8 weeks after the treatment begun, earlier than that from 6 to 8 weeks after the beginning of interferon alone in the literature.

CONCLUSIONSequential antiviral therapy had much higher rates of long-term HBeAg seroconversion, undetectable HBV DNA and normalization of alanine aminotransferase with good cost-effectiveness. Its mechanism to promote the antiviral effect might be dependent on the immunoregulatory action of thymosin alpha1 in the earlier period and the specific inhibition of HBV DNA replication by lamivudine in the later period of the therapeutic course.

Adjuvants, Immunologic ; administration & dosage ; Antiviral Agents ; administration & dosage ; China ; Drug Therapy, Combination ; Hepatitis B, Chronic ; drug therapy ; Humans ; Interferon-alpha ; administration & dosage ; Lamivudine ; administration & dosage ; Thymosin ; administration & dosage ; analogs & derivatives ; Treatment Outcome

OBJECTIVETo optimize cultivation methods of bone marrow mesenchymal stem cells (MSCs) from hepatitis B patients and to investigate their biological characteristics.

METHODSGrowth curves of hepatitis B patients MSCs cultivated with five culture media and two inoculation methods were compared; the shapes, appearances, surface markers and bionomics of the cultivated MSCs were studied.

RESULTSInoculating the cells obtained directly from bone marrow aspirations was not as successful as using the marrow cells after their density gradient centrifugations (76% vs 88%), but the differences in the results were not statistically significant (P more than 0.05). The successful cultivation rates using five culture media were different and the differences were statistically significant (P less than 0.01). The autoserum medium was most successful, fatal bovine serum (FBS) medium was next successful and the non-patient serum medium was the least successful. The growth curves of the cultivations using the different media were parallel to this. Changing the whole culture media every 2 or 3 days was better than changing half of the media. The shapes, appearances, surface markers and the growth characteristics of MSCs from the hepatitis B patients were almost the same as MSCs from the normal adult.

CONCLUSIONThe best cultivation method of MSCs from hepatitis B patients is: separating marrow cells using density gradient centrifugal separation, cultivating them using an autoserum culture medium, and completely changing the medium every 2-3 days. The biological characteristics of MSCs from the hepatitis B patients using the above methods are almost the same as those from normal adults.

Adult ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cells, Cultured ; Culture Media ; Hepatitis B ; Humans ; Mesenchymal Stromal Cells ; cytology ; Middle Aged

OBJECTIVETo study the relationship between a G/T substitution at position -88 of myxovirus resistance-1 gene (MxA) and the self-limiting or chronic infection of HBV.

METHODSBlood samples from 100 patients with self-limiting HBV infection (positive anti-HBs and anti-HBc) and from 340 patients with chronic HBV infection were collected. MxA-88 G/T polymorphism was typed using a protocol based on competitively differentiated-polymerase chain reaction. For statistical analysis, odds ratio and chi-square test were used.

RESULTSThe detective rate of G/G genotype (low expression genotype) of MxA-88 G/T was 50.2% (221/440), those of T/T genotype (high expression genotype) and G/T heterozygous genotype were 5.5% (24/440) and 44.3% (195/440). Compared to patients with chronic infection, patients with self-limiting infection had lower frequency of G/G genotype (41.0% vs 52.9%, P < 0.05) or G allele (62.5% vs 75.9%, P < 0.01) and had higher frequency of T/T genotype (16.0% vs 2.4%, P < 0.01) or T allele (37.5% vs 24.1%, P < 0.01), but there was no significant difference in the G/T heterozygous genotype.

CONCLUSIONSMxA gene -88 G/T polymorphism influences the natural outcomes of HBV infection to some extent. This SNP of MxA gene may be used as a clinical prognostic marker of HBV infection.

Adult ; Biomarkers ; Female ; GTP-Binding Proteins ; biosynthesis ; genetics ; Genotype ; Hepatitis B, Chronic ; genetics ; Humans ; Male ; Myxovirus Resistance Proteins ; Polymorphism, Single Nucleotide ; genetics ; Prognosis

OBJECTIVESTo probe into the initiative factors of the damage sensitive stage of hepatocytes induced by interferon in patients with chronic hepatitis B (CHB).

METHODSForty-four CHB patients with positive HBeAg and HBV DNA were treated with interferon. Serum ALT and viral markers levels of HBsAg, HBeAg, anti-HBc and HBV DNA were examined regularly. Liver biopsy was carried out just before the treatment.

RESULTSThe rate of HBeAg seroconversion was 75% at the sixth month, and 68.2% after one year of follow up. The rate of damage sensitive stage of hepatocytes was 47.7%. The average onset time was (3.14+-1.49) weeks after the treatment, and lasted for (8.24+-3.52) weeks. The ALT level raised (1.73+-1.13) times. The occurrence of damage sensitive stage of hepatocytes was indicator for good curative effect (Fisher exact probability, P=0.028). Damage sensitive stage of hepatocytes was more often developed in patients with moderate inflammation, overexpression of HBcAg in liver and higher level of HBeAg in blood stream before treatment. HBeAg and anti-HBc levels in peripheral blood decreased in the onset period of damage sensitive stage of hepatocytes.

CONCLUSIONSThe initiative factors of the damage sensitive stage of hepatocytes may be: HBeAg decreasing in peripheral blood induced by interferon may dismiss immune lutation of HBeAg and anti-HBc to cytotoxic T lymphocyte (CTL), which recognize HBcAg as target, thus activates the cytotoxicity of HBV-infected hepatocytes mediated by CTL.

Adolescent ; Adult ; Alanine Transaminase ; blood ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; metabolism ; pathology ; Humans ; Interferon-alpha ; adverse effects ; therapeutic use ; Liver ; pathology ; Male ; Middle Aged ; T-Lymphocytes, Cytotoxic ; drug effects

OBJECTIVETo investigate the cleavage of HBV core protein in vivo by proprotein convertase furin or its family members and observe the intracellular localization of the putative cleaved product.

METHODSRecombinant HBV core protein was incubated with furin under different conditions in vitro, and the reaction was checked with Western blotting. The recombinant vectors expressed the putative cleaved fragment and intact core protein (serves as control) were constructed. The stable expression cell lines were established by transfecting constructs into HepG2 cell line, for which indirect immunofluorescence staining was used by monoclonal anti-HBc against the region shared by core protein and its cleaved product .The confocal microscopy was carried out to observe the intracellular distribution.

RESULTSHBV core protein was cleaved by furin in vitro under different tested conditions. The molecular weight of the major cleaved product just about 15,000 was in concordance with the expectation. The expressed cleaved fragment could react to the monoclonal antibody against core protein, and mainly located in cytosol in particle style just like the intact core protein.

CONCLUSIONHBV core protein can be cleaved by furin in vitro. The major cleaved product has similar antigenicity and subcellular distribution to core protein. These data suggest that proprotein convertase furin or its family members play important roles in HBV replication regulation, and the cleaved product may be involved in antiviral immunity of HBV infection. Further investigations are imperative.

Furin ; metabolism ; Genetic Vectors ; Hep G2 Cells ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B virus ; metabolism ; physiology ; Humans ; Microdissection ; Microscopy, Confocal ; Proprotein Convertases ; metabolism ; Transfection ; Virus Replication

Objective: To investigate the possibility of adipose tissue mesenchymal stem cell-derived microvesicles (ADSC-MVs) to improve the retention volume of fat transplantation. Methods: Human adipose tissue was obtained from 5 healthy female patients aged from 20 to 30 years, who came to the hospital for abdominal liposuction. Adipose tissue mesenchymal stem cells (ADSCs) were acquired by collagenase enzymatic hydrolysis. ADSC-MVs were isolated from the supernatant of cultured ADSCs through ultra-centrifugation, and characterized by transmission electron microscope and PKH26 staining. Sixteen BALB/c-nu nude mice were randomly divided into 2 groups (n=8, mice/group) by random number table method. In EV group, the mice were subcutaneously injected 0.35 ml human fat, with 7 μg ADSC-EVs (final concentration is 20 μg/ml), while 0.35 ml human fat were in blank group. The mice were sacrificed and the grafts were harvested at 1 month and 3 months after transplantation. Weight and volume of transplanted fat were tested. Morphology of adipocytes, fibrosis and necrosis of fat tissue were confirmed by HE staining. Blood vessel density were accessed by CD31 staining.SPSS 13.0 was used for statistical analysis and Student-t test was applied to compare the data from two groups. Results: One month after fat transplantation, the volume of fat grafts in blank group was lower than that in MV group. Besides, lipoliquefaction can be observed in fat grafts of blank group. Fat grafts in MV group had a higher volume and littler necrosis and lipolique faction. Three months after grafting, the volume of fat grafts in blank group [(0.057±0.009) ml] was significantly lower than that in MV group [(0.103±0.015) ml], t=8.117, P<0.001. MV group had an increased number of vessels (29.6±4.0/field) compared with grafts from the Blank group (23.0±2.4/field), t=2.825, P=0.022. Histological analysis revealed that the fat grafts in MV group consisted less fat necrosis and fibrosis than control group. Conclusions ADSC-MVs can improve the retention volume of fat transplantation.

Objective To explore the effect of edaravone (ED) on the neurological functionaldeficits, apoptosis and expression of caspase-3 protein following focal cerebral ischemia/reperfusion(I/R)injury in rats. Methods A total of 24 male Sprague-Dawley (SD) rats were randomly allocated intothe sham-operation group, cerebral I/R group, normal saline treatment group and ED treatment group, 6rats in each group. Rat models with focal cerebral I/R injury induced by middle cerebral artery occlusion(MCAO) were established using a modified suture method. ED (3mg/kg) or equal volume of normalsaline was injected intraperitoneally immediately after cerebral ischemia and 12 h after reperfusion in thetreatment groups;the rats in sham-operation group underwent the same modeling procedure withoutischemia by nylon suture. The neurological behavioral deficits were evaluated 24 h after I/R injury;,immunohistochemical staining and Western blot assay were applied to detect the change in the expressionof caspase-3 protein; in situ TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay was used tostudy the change in neuronal apoptosis. Results The scores of neurological behavioral deficit scale,the positive cells and expression of caspase-3 protein, and the apoptotic cells in the ED treatment groupwere significantly decreased, compared with that of the I/R group and normal saline treatment group(P<0.05 for each comparison). Conclusion ED may effectively reduce neuronal apuptosis andneurological functional deficits after cerebral I/R injury, which might be related with the inhibition of thecaspase-3 protein expression.

OBJECTIVETo detect HBV antigen specific cytotoxic T lymphocyte (CTL) changes in patients during acute flare-ups and to study their association with flare-ups and aggravations into grave hepatitis by quantitative analysis of HLA-A2* restricted HBcAg-specific CTL cells.

METHODSThe frequency of HBcAg-specific CTL cells in the peripheral blood mononuclear cells (PBMC) from 29 patients with persistent infection with HBV were quantified by flow cytometry using one HLA-A2*HBV peptide pentamers complex (Pro5TM MHC Pentamers).

RESULTSThere was a statistical difference of HBcAg specific CTLs between the patients with acute exacerbations (1.4%+/-0.8%) and the patients with immune tolerance (0.6%+/-0.4%) (t = 2.180, P = 0.01-0.05); There was no significant difference between the grave hepatitis group (1.3%+/-1.0%) and the chronic hepatitis group (1.4%+/-0.8%) regarding frequencies of antigen specific CTL (t = 0.215, P = 0.833-0.05). The level of antigen specific CTLs in PBMC in the 6 cases of chronic hepatitis B with acute exacerbations maintained a relatively high level (more than 0.7%) within the 12 week follow-up period.

CONCLUSIONHBcAg-specific CTLs may play an important role in hepatic flare-ups in patients with chronic HBV infection, but there was no direct relationship between antigen- specific CTLs and grave hepatitis.

Adult ; Female ; HLA-A2 Antigen ; immunology ; Hepatitis B Core Antigens ; immunology ; Hepatitis B virus ; immunology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Male ; Middle Aged ; T-Lymphocytes, Cytotoxic ; immunology ; Viral Load ; Young Adult