Objective To analyze the feature of B cell epitope of soluble liver antigen(SLA) in autoimmune hepatitis. Methods Twenty-one autoimmune hepatitis(AIH) were enrolled the study. The control groups included 15 primary biliary cirrhosis(PBC) patients, 20 chronic virus hepatitis patients and 20 heaths. The serum incubated with coated one dimension of SLA peptide pools by ELISA for mapping the B cell epitopes. ELISPOT was performed to screen the SLA specific T cell response by observing the secretion of IFN-γ after PBMC simulated. Results Ten serum samples from AIH patients recognized the SLA peptide pools, the frequency was 47.62%, which was higher than PBC group, virus hepatitis group and normal control group, P value was 0.001, <0.001 and <0.001, respectively. The 10 positive serum dominant response to SLA 7 th pool(aa385-441), which laid in the unique no conservative region of the SLA molecular. T cell recognizing regions laid dominantly in first and second pool, the frequency was 30% and 40%, respectively, however the recognized frequency of 7th pool was 10%. Conclusion B cell linear epitopes lie in the sequence aa385-441, which is the unique unconservative region in the high conservative SLA molecular and overlaps with part sequence of T cell epitopes. The results show it is complex and interaction that SLA induced humoral-mediated immunity and cellular immunity to involve in AIH pathogenesis.

OBJECTIVEDetection HBV DNA among HBsAg negative children who have been vaccined at birth, in order to improve the evaluation of the indicator for HBV DNA infection.

METHODSSelection HBsAg negative children who have been vaccined at birth and then detection HBV DNA from sera using QIAamp Viral DNA Mini Kit, HBV DNA s region was obtained by nested PCR and sequencing.

RESULTS12 of the 140 children were HBV DNA detected were positive and the infectious rate was 8.6% . No mutant of the 12 HBV DNA in "a" determinant.

CONCLUSIONTo evaluate the effection of the prevention of HBV mother-to-child transmission, the standard method should be established. The detection of HBV DNA should be included in the future.

Child ; Child, Preschool ; DNA, Viral ; blood ; genetics ; Female ; Hepatitis B ; blood ; prevention & control ; transmission ; Hepatitis B Surface Antigens ; blood ; genetics ; Hepatitis B virus ; genetics ; Humans ; Infectious Disease Transmission, Vertical ; prevention & control ; Male ; Mutation

Fosmidomycin (100 micromol x L(-1)) which is the effective inhibitor of DXR, key enzyme in terpenoid MEP pathway, was used to treat with hairy roots of Salvia miltiorrhiza. The treated roots were harvested at 2, 4, 6, 8, 10, 16 and 21 d, mRNA level of SmDXR and tanshinone content in treated and negative control groups were detected. Results found that, after treated with fosmidomycin, color of S. miltiorrhiza hairy roots grew pale gradually comparing with controls; mRNA level of SmDXR in hairy roots varied as a shape of parabolic and the highest value achieved at the sixth day after treatment, then it decreased gradually; Content of four kinds of tanshinones were detected. Among of the four kinds of tanshinones, Tanshinone I content changed relatively little, while content of dihydrotanshinone I, cryptotanshinone and tanshinone II (A) decreased gradually in 21 days. The content of total tanshinones in NC groups was 5, 63 times more than FOS-treated roots in the 21th day. The previous results showed that SmDXR played an important role in the accumulation of tanshinone content in MEP pathway. Once the mRNA level of SmDXR was suppressed, the accumulation of secondary metabolites will be significantly affected.
Aldose-Ketose Isomerases ; genetics ; Diterpenes, Abietane ; metabolism ; Fosfomycin ; analogs & derivatives ; pharmacology ; Gene Expression Regulation, Plant ; drug effects ; Plant Roots ; drug effects ; growth & development ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; growth & development ; metabolism ; Time Factors

OBJECTIVETo explore the effect of Modified Hangqi Chifeng Decoction (MHCD) on levels of collagen type IV (Col IV), matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2) in extracellular matrix (ECM) of glomerular mesangial cells (GMCs) in LPS induced mice.

METHODSNormal serum and telmisartan, high, medium, low dose MHCD containing serums were prepared by using serum pharmacology method. GMCs were cultured in vitro. The proliferation of mesangial cells were induced using LPS as stimulating factor. GMCs were divided into six groups, i.e., the normal group, the model group, the telmisartan group, high, medium and low dose MHCD groups. Col IV content in the supernatant of mesangial cells was detected using ELISA. Protein expressions of MMP-2 and TIMP-2 were detected using Western blot.

RESULTSCompared with the normal group, Col IV content obviously increased in the model group after 72-h LPS stimulation; protein expressions of MMP-2 and TIMP-2 were obviously up-regulated, and MMP-2/TIMP-2 ratio was down-regulated in the model group (P < 0.01). Compared with the model group, Col IV content obviously decreased in high and medium dose MHCD groups and the telmisartan group (P < 0.01); protein expressions of MMP-2 were obviously down-regulated in medium and low dose MHCD groups (P < 0.01, P < 0.05); the protein expression of TIMP-2 was obviously down-regulated in high, medium, low dose MHCD groups and the telmisartan group (P < 0.01). The pro- tein expression of TIMP-2 was obviously lower in the high dose MHCD group than in the low dose MHCD group (P < 0.01). MMP-2/TIMP-2 ratio was obviously up-regulated in the telmisartan group, high and medium dose MHCD groups (P < 0.01).

CONCLUSIONMHCD could regulate disordered MMP-2/TIMP-2 ratio in LPS induced ECM, inhibit excessive production of Col IV in ECM, promote the degradation of ECM, reduce the accumulation of ECM, thereby, delaying the process of glomerular sclerosis.

Animals ; Cells, Cultured ; Collagen Type IV ; metabolism ; Extracellular Matrix ; metabolism ; Kidney Glomerulus ; cytology ; Matrix Metalloproteinase 2 ; metabolism ; Mesangial Cells ; drug effects ; Mice ; RNA, Messenger ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

The chemical constituents from Euphorbia lunulata was investigated in this paper. Fourteen compounds were isolated and purified by column chromatographies on silica gel and preparative HPLC. Their structures were identified by physiochemical properties and NMR data analysis as lupeol (1), euphol (2), cassipourol(3) , 24-methylenecycloartan-3beta-ol (4), 24-hydroperoxycycloart-25-en-3beta-ol (5), 25-hydroperoxycycloart-23-en-3beta-ol (6), betulin (7), uvaol (8), (23E) -25-methoxycycloart-23-en-3beta-ol (9), (23E) -cycloart-23,25-dien-3beta-ol (10), 24-methylenecycloartan-3beta, 28-diol (11), salicinolide (12), 2alpha, 3beta, 5alpha, 9alpha, 15beta-pentaacetoxy-11,12-epoxy-7beta, 8alpha-diisobutyryloxyjatropha-6 (17) -en-14-one (13) and 3beta, 5alpha, 15beta-triacetoxy-7beta-isobutyryloxy-9alpha-nicotinoyloxyjatropha-6 (17), 11(E)-dien-14-one (14). Among them, compounds 1-11 were isolated from E. lunulata for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Euphorbia ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Stereoisomerism

Objective To detect the volume changes of cerebral gray and white matter in patients with maintaining hemodialysis using voxel-based morphometry(VBM) and to correlate these changes with cognitive function. Methods Forty-two patients with maintaining hemodialysis and 41 age and sex matched normal subjects were recruited in this study. MMSE was obtained to evaluate their neuropsychiatric conditions. Whole brain high-resolution T1WI was performed on 3.0 T MR scanner in both patients and normal controls. The data were analyzed by VBM based on SPM8, using analysis of functional neuroimages (AFNI) software package with the Monte Carlo simulation method(AlphaSim method) for multiple cluster level comparisons correction. Independent sample t test analysis was used to compare the volume of gray and white matter between the patients and normal controls. In addition, Stepwise multiple linear regression analysis was performed to explore the correlation between the voxel value of cerebral volume changes area and dialysis duration and clinical laboratory examination, and Spearman correlation analysis was used for the correlation between the left insula voxel values and neuropsychological test scores. Results Compared with normal controls, patients showed significantly decreased volume in the grey matter of the right putamen, the left putamen, the left insula (numbers of voxel in clusters were 455, 561, 162, t=-9.5681,-5.9516,-5.7185,P<0.001, AlphaSim-corrected). There was negative correlation between decreased grey matter volume of the right putamen[(0.53 ± 0.12)mm3]and the left putamen[(0.48 ± 0.12)mm3] and dialysis duration [19.0(1.5-114.0)months] (r=-0.330,-0.307,P<0.05). MMSE score of patients[29(21-30)score] was significantly lower than normal controls[30(28-30)score] (Z=-30.58,P<0.01). Decreased grey matter volume of the left insula [(0.39 ± 0.12) mm3] was positively correlated with MMSE(r=0.320, P<0.05). Conclusions The patients with maintaining hemodialysis show grey matter atrophy which is associated with neurocognitive dysfunction. Dialysis duration may be an important risk factor for decreased gray matter in patients with maintaining hemodialysis.

Objective To evaluate the efficacy,safety and impact of recombinant human cytotoxic T lymphocyte-associated antigen (CTLA)-4 fusion proteins (rhCTLA-4Ig) on serum human tumor necrosis factor (TNF)-α and CX3CL1 in active rheumatoid arthritis (RA) patients.Methods Forty-four RA patients were treated with rhCTLA-4Ig and placebo.Clinical response was assessed by American College of Rheumatology (ACR) criteria and disease activity score in 28 joints (DAS28).The levels of serum TNF-α and CX3CL1 were determined in 44 RA patients and 20 healthy controls by enzyme-linked immunosorbent assay (ELISA).Comparisons between groups were performed by t-test or x2 test.Results At week 12,ACR20,ACR50and ACR70 responses in RA patients with rhCTLA-4Ig were achieved by 95％(20/21 ),76％( 16/21 )and 19％(4/21) respectively,but no patient with placebo achieved ACR20,ACRS0 and ACR70 responses.There were significantly statistical differences in ACR20 and ACR50 responses (x2=39.17,26.69,P＜0.01 ).At week 12,the mean DAS28 in the rhCTLA4Ig group was 3.1±1.3 versus 6.2±1.1 at baseline (P＜0.01).Similarly,health assessment questionnaire (HAQ) improved significantly,declining from 1.4±0.5 at baseline to 0.4±0.5 at week 12 (P＜0.01).However,the mean DAS28 in the placebo group was 5.8±1.2 versus 6.0±0.7 at baseline (P＞0.05),HAQ declined from 1.6±0.4 to 1.6±0.6 (P＞0.05).In addition,there were higher levels of TNF-α and CX3CL1 in the active RA patients than those of the healthy controls (P＜0.01).After 12 weeks therapy,Serum TNF-α and CX3CL1 levels in the rhCTLA-4Ig group decreased significantly (P＜0.01).There weren't decline in the placebo group (P＞0.05).Conclusion This study has shown that rhCTLA-4Ig is very effective in reducing disease activity,improving function during the 12 weeks treatment.rhCTLA-4Ig therapy for 12 weeks can lead to significant decrease of serum TNF-α and CX3CL1.

Objective To evaluate the distribution and age-related changes of brain iron content in healthy people using MR quantitative susceptibility mapping (QSM).Methods Sixty three healthy right-handed volunteers underwent the routine MRI and SWI scan to get SWI-unfiltered phase images and magnitude images.QSM were reconstructed from the SWI-unfiltered phase images and magnitude images using SMART software.The regions of interest at the bilateral frontal white matter and nuclei (caudate nuclews,globus pallidus,putamen,substantia nigra,dorsal thalamus,red nucleus and dental nucleus) were drawn manually,and the susceptibility was measured.The linear correlation between the susceptibility and iron concentration cited from Hallgren and Sourander's post-mortem brain study was calculated.Wilcoxon test were applied to calculate the difference between the bilateral frontal white matter and bilateral nuclei.The correlation of age with susceptibility of bilateral frontal white matter and bilateral nuclei were analyzed by Spearman correlation analysis.Results The median susceptibility (extent) of frontal white matter,caudate nucleus,globus pallidus,putamen,dorsal thalamus,substantia nigra,red nucleus and dentate nucleus of 63 healthy people were-12.81 (-31.56,8.72),39.27 (22.35,75.13),93.99 (19.19,158.75),29.16 (4.11,81.53),2.91 (-27.80,27.95),83.14 (38.57,185.79),63.49 (13.83,142.09),63.30 (36.78,128.53) ppb (× 10-9),respectively.There was high consistency with Hallgren and Sourander's study (r=0.91,P＜0.05).The susceptibility of globus pallidus was the highest,followed by substantia nigra.The least susceptibility was seen in the frontal white matter.The susceptibility of right caudate nucleus,substantia nigra,red nucleus and dental nucleus was higher than that of left side (Z value was-3.18,-4.44,-3.70 and-2.64,respectively,P＜0.05).The susceptibility of bilateral globus pallidus of the male was significantly different from that of the female (Z value was-2.27 and-2.42,respectively,P＜0.05).There was positive correlation between age and the susceptibility of bilateral caudate nucleus,putamen,red nucleus and dental nucleus (r value was 0.30 to 0.49,P＜0.05).Conclusions QSM based on the SWI-unfiltered phase and magnitude images can clearly display the cerebral nuclei and evaluate the brain iron content accurately,which is consistent with the histopathological results.Iron content of bilateral caudate nucleus,putamen,red nucleus and dental nucleus increase with aging.

To investigate the clinical efficacy and safety of phototherapeutic keratectomy ( PTK ) in the treatment of bullous keratopathy ( BK) .METHODS: A retrospective analysis of 60 cases ( 60 eyes) of BK patients from Department of Ophthalmology in our hospital October 2011 to July 2014 was undergone. Clinical data of all patients with treatment of PTK were analyzed. Best corrected visual acuity ( BCVA ) , corneal curvature, corneal astigmatism, corneal thickness and corneal endothelial cell density ( ECD ) , postoperative complications before and after surgery were compared. RESULTS: BCVA, corneal curvature, corneal astigmatism of patients before surgery were 0. 05 ± 0. 01 and 37. 02±5. 38, 1. 08±0. 67D, which were significantly less than those of postoperative ( respectively 0. 45 ± 0. 13 and 46. 27 ± 7. 02, 1. 92 ± 0. 73D ), the differences were statistically significant (all P<0. 05). Corneal thickness of patients was 492. 33 ± 18. 27μm before surgery, which was higher than that after surgery 377.27±22.49μm (P<0.05). The difference of visual acuity before and after surgery was statistically significant in this group (P<0. 05). During the follow-up period of 6mo, no recurrence of the original corneal lesions, only 2 cases of postoperative slight haze, it was completely dissipated after given the hormone eye drops.CONCLUSlON: Excimer laser technology has high safety in the treatment of bullous keratopathy, it should be promoted in clinical practice.

Objective To explore the responses of antigen-specific T cells stimulated by hepatitis B virus(HBV)-specific proteins in chronic hepatitis B patients accepting antiviral therapy. Methods Seventeen patients with chronic hepatitis B (CHB) accepting antiviral therapy were included in this study. The peripheral blood monocular cell ( PBMC) were separated from the whole blood collected at the three different time of before and one and three months after accepting antiviral therapy. ELISPOT assay was used to detect the frequency and strength of secreting IFN-γ cells of PBMC stimulated by HBsAg, HBcAg and HBeAg. HBV virus loading, HBsAg, HBeAg, ALT and AST in serum were detected at the same time. Results After three months therapy, ALT, TBiL were improved in all patients, and HBV DNA level were dropped and undetectable in 11 cases. The rates of T cell response in patients to HBV specific proteins were 64. 7% , 76. 5% and 82. 4% at the time of before and one and three months after accepting antiviral therapy, respectively. The frequency of responses of antigen-specific T cells stimulated by HBcAg was higher than that stimulated by HBsAg or HBeAg, and the frequency was enhanced after antiviral therapy. The average response magnitude was expressed as spot forming cells (SFC) per million input cells. SFC of T cell responses to HBcAg was also higher than to HBsAg or HBeAg. There was no significant difference in SFC of T cell responses to HBsAg or HBeAg at the time of before and after antiviral therapy, but there were significant difference in SFC of T cell responses to HBcAg at the time of before and after antiviral therapy. SFC of T cell responses to HBcAg was negatively associated with HBV DNA, and no associated with level of ALT in serum. Conclusion The responses of antigen-specific T cells were improved in CHB patients accepting antiviral therapy which associated with the decrease of HBV DNA. It suggested to investigate HBV specific T cell responses was important.