Objective To investigate the association of cytotoxic T lymphocyte associated antigen 4 (CTLA 4) gene polymorphism with type 1diabetesmellitusandautoimmunethyroiddiseases. Methods The A/G phenotype at position 49 of the CTLA 4 gene exon 1 was determined by PCR RFLP method in 33 classic type 1 diabetes patients, 57 latent autoimmune diabetes in adults (LADA) patients, 122 autoimmune thyroid disease patients and 84 healthy control subjects of Chinese Han. Results The frequency of the CTLA 4 G phenotype was significantly higherintype1diabetespatientsthanthatincontrol subjects (55.6% vs 36.9%, P=0.0005). Neither the presence nor the absence of G allele influenced the occurrence of islet autoantibody (ICA) and glutamate decarboxylase antibody (GADA). The strong association of the CTLA 4 G allele with AITDs was showed in our study (66.4% vs 36.9%, P

Cardiac pacing site has an important impact on the activation sequence and systolic synchrony of heart , which is an important factor determining clinical effect of cardiac pacing .Along with expanding and use of active fixed electrode in clinic ,especially adjustable bending delivery sheath ,it makes right atrial pacing of non -tradition‐al site possible .But whether changing atrial pacing site can reduce atrial fibrillation load after pacemaker implanta‐tion is still controversial .Now the present article made a simple review about research progress of interatrial septum pacing .

Objective　To investigate the association of gene polymorphism of cytotoxic T lymphocyte-associated antigen 4（CTLA-4） with autoimmune thyroid diseases. Methods　The A／G transition polymorphism at position 49（exon 1，codon 17） of the CTLA-4 gene was determined by polymerase chain reaction restriction fragment length polymorphism （PCR-RFLP）method in 122 autoimmune thyroid diseases patients which included 87 Graves’ disease （GD） patients and 35 Hashimoto's thyroiditis（HT） patients， as well as 84 control subjects. We detected their thyroid function by ELISA technique， and the thyroid autoimmune antibodies （TGAb，TPOAb） by indirect immunofluorescent technique. Results　The strong association of the CTLA-4／G49 allele with AITDs was seen in our study（66.4％ vs 36.9％ P＜0.0001）. The G allele in GD patients was significantly increased compared with control subjects（69.5％ vs 36.9％， P＜0.0001）. In HT patients， the frequency of G allele was also higher than control subjects（58.6％ vs 36.9％，P＜0.01）， and there was no significant difference between HT and GD groups. When GD and HT subjects were stratified with respect to sex， neither female nor male patients demonstrated evident association of G49 allele with gender.Conclusions　The polymorphism of CTLA-4 gene （exon 1 condon 17 position 49）confers susceptibility to AITDs. This association is independent of sex.

Objective To increase the killing effect of aminolevulinic acid (ALA)-based photodynamic therapy (PDT) on a human skin squamous cell carcinoma cell line A431 by poly lactic-co-glycolic acid nanoparticles (PLGA NPs).Methods ALA-loaded PLGA NPs (ALA PLGA NPs) were prepared using a modified double emulsion solvent evaporation method,and characterized in terms of size,encapsulation efficiency,loading capacity and morphology.Transmission electron microscopy was carried out to observe the morphology of A431 cells after uptake of ALA PLGA NPs.To optimize incubation time,multi-mode microplate reader was used to describe the fluorescence kinetics of protoporphyrin Ⅸ generated by A431 cells during 24 hours of incubation with 0.1 mmol/L ALA,1 mmol/L ALA,2.7 g/L ALA PLGA NPs containing about 0.1 mmol/L ALA,and PLGA NPs without ALA separately.Some A431 cells were divided into 10 groups:control group receiving neither treatment nor irradiation,0.1 and 1 mmol/L ALA dark/PDT group incubated 0.1 and 1 mmol/L ALA respectively,ALA PLGA NP dark/PDT group incubated with ALA PLGA NPs of 2.7 g/L,PLGA NP dark/PDT group incubated with PLGA NPs of 2.7 g/L,simple irradiation group irradiated with a He-Ne laser (wavelength:635 nm,power density:8.6 mW/cm2,energy density:8 J/cm2) only.The dark groups were kept in darkness strictly by wrapping in aluminum foil,and PDT groups were irradiated using a He-Ne laser.After another 24 hours of culture following irradiation,methyl thiazolyl tetrazolium (MTF) assay was conducted to estimate the survival rate of cells.To study the effect of ALA PLGA NPs on cell apoptosis,some A431 cells were divided into three groups:control group receiving neither treatment nor irradiation,ALA-PDT group and ALA PLGA NP PDT group receiving PDT after incubation with ALA and ALA PLGA NPs respectively.Flow cytometry was performed to detect the apoptosis of A431 cells at 12 and 24 hours,separately,after the photodynamic therapy.Data were statistically analyzed using SPSS 13.0 software package by means of a t test.Results The prepared ALA PLGA NPs were spherical with a mean particle size of (65.6 ± 26) nm,encapsulation efficiency of (65.8 ± 7.2) ％,and drug loading capacity of (0.62 ± 0.27)％.ALA PLGA NPs could be uptaken by A431 cells and gathered in the cytoplasm.The PpIX fluorescence kinetic study showed that the fluorescence intensity increased with time within 24 hours in A431 cells incubated with ALA or ALA PLGA NPs.After 6 and 24 hours of incubation,the A431 cells incubated with 2.7 g/L ALA PLGA NPs showed a significant increase in the protoporphyrin Ⅸ fluorescence intensity compared with those incubated with 0.1 mmol/L ALA (both P ＜ 0.01).Further more,the survival rate of A431 cells was statistically lower in the ALA PLGA NP PDT group than in the 0.1 mmol/L ALA PDT group at 6 and 24 hours (t =35.685,5.262,respectively,both P ＜ 0.01).Elevated apoptosis rate was observed in the ALA PLGA NP PDT group compared with the ALA PDT group at 12 ((13.10 ± 0.50)％ vs.(4.90 ± 0.13)％,t =9.074,P＜ 0.01) and 24 ((30.17 ± 1.02)％ vs.(11.6 ± 0.59)％,t =9.095,P ＜ 0.01) hours.Conclusions ALA PLGA NPs can promote the formation of protoporphyrin Ⅸ,strengthen the killing effect of ALA-PDT on A431 cells in vitro,and enhance the apoptosis induced by ALA-PDT in tumor cells.

Objective To explore the expression of Tau protein in brains of the intractable epilepsy patients and discuss its function in the pathogenesis. Methods Immunohistochemistry was adapted to test the expression of Tau protein and the mossy fiber sprouting in the hippocampus and cortex of temporal lobe from 48 intractable epilepsy patients and age-mached 48 cases of controls. Results The expression of phosphorylated Tau protein in the CA_3 areas (0.0450?0.0115) and the molecular layers of dentate gyrus (0.0463?0.0120) in intractable epilepsy patients was increased, accompanied by the hippocampus mossy fiber sprouting (3.18?0.35). No difference on the expression of total Tau protein was observed between the intractable epilepsy patients and the controls. Conclusion The present findings demonstrate that the strengthening of phosphorylated Tau protein might play an important role for the synaptic reorganization of the hippocampus. Thus, the changes of phosphorylated Tau protein could improve clinical prognosis in the intractable epilepsy patients.

A beta-D-xylosidase from Leifsonia shinshuensis DICP 16 was purified to apparent homogeneity using a combination of ammonium sulfate precipitation, DE 52 anion-exchange, Q-Sepharose Fast Flow anion-exchange, Toyopearl Butyl 650C hydrophobic-interaction and Sephacryl S-300 HR gel-permeation chromatography. The purified xylosidase consisted of two same subunits and had the relative molecular weight of 180 kD as determined by SDS-PAGE and gel-permeation chromatography. The maximal beta-D-xylosidase activity occurred at 55 degrees C and pH 7.0. It was stable at 45 degrees C and retained its original activity for 60 min. The stability declined rapidly when the temperature rose above 55 degrees C. The xylosidase was stable in the pH range from 6.0 to 11.0 for 20 h. At pH 7.0 and 45 degrees C the Km for p-nitrophenyl-beta-D-xylopyranoside (pNPX) was 1.04 mmol/L and the Vmx was 0.095 mmol nitrophenol/min/mg xylosidase. The enzyme was inhibited strongly by Fe2+ and Cu2+. It exhibited low levels of activity against other artificial substrates, compared to its activity against pNPX. When different natural xylosides were used as the substrates, the xylosidase showed distinct hydrolysis ability. It could hydrolyze 20-C, beta-(1-->6)-xyloside of ginsenoside Rb3 (G-Rb3) into ginsenoside Rd, but did not hydrolyze the other beta-D-glucosidic bonds of G-Rb3. Additionally, the xylosidase could not hydrolyze C-7 xylosyl-bearing taxanes.
Actinomycetales ; classification ; enzymology ; Amino Acid Sequence ; Culture Media ; Hydrogen-Ion Concentration ; Molecular Sequence Data ; Sequence Analysis, Protein ; Temperature ; Xylosidases ; chemistry ; genetics ; isolation & purification

Objective:To study the expression of ULBP2 protein in the brain tissues of patients with drug-refractory epilepsy and its clinical significance.Methods:Gene-chip,immunofluorescence and Western blot were used to test expression of ULBP2 in the surgically removed brain tissue of patients with drug-refractory epilepsy from the brain bank of our department(n=42),and the results were compared with that of normal controls (n=12).Results:The relative increasing expression of ULBP2-gene in the brain of patients with drug-refractory epilepsy,and ULBP2 protein expression was significantly increased in temporal lobe cortex of patients with drug-refractory epilepsy as compared with the same regions of the controls specimens.Conclusion:The results indicate that the overexpression of ULBP2 may be involved in the pathophysiology of drug-refractory epilepsy.

Background Equivalent continuous A-weighted sound pressure level is not appropriate for evaluating the risk of non-steady noise exposure, and need to be corrected by noise time-domain structure, but the correction method and its applicability need to be discussed. Objective To validate the application of the kurtosis-adjusted normalization of equivalent continuous A-weighted sound pressure level to a normal 8 h working day ( L_{Aeq,8 h}) in assessing noise-induced hearing loss (NIHL), and to improve the methods for assessing occupational hearing loss associated with different types of noise. Methods Audiometric and shift-long noise exposure data were acquired from a population(n=2 466) of screened workers exposed to noise between 70 dB(A) and 95 dB(A) from 6 industries in China. The cohort data were collapsed into 1 dB(A) bins, and the average kurtosis and noise-induced permanent threshold shifts at 3 kHz, 4 kHz, and 6 kHz (NIPTS₃₄₆) within 1 dB(A) were calculated respectively. According to the existing correction method, the adjustment coefficient λ was calculated by multiple regression, and L_{Aeq,8 h} was corrected by λ (L'_{Aeq,8 h}). The entire cohort was divided into K₁ (≤10; steady noise), K₂ (10~50; non-steady noise), and K₃ (＞50; non-steady noise) groups based on mean kurtosis levels. Predicted NIPTS₃₄₆ was calculated using the ISO 1999 model for each participant and the actual measured NIPTS₃₄₆ was corrected for age and gender. The underestimated NIPTS₃₄₆ was the difference between the values of estimated NIPTS₃₄₆ and the corresponding actual NIPTS₃₄₆. To validate the applicability of L′_{Aeq,8 h} in evaluating NIHL, the correlation between L′_{Aeq,8 h} and HFNIHL, and the mean difference between real NIPTS₃₄₆ and estimated NIPTS₃₄₆ were analyzed. Results The adjustment coefficient λ was determined at 5.43. The results of multiple logistic regression analysis showed that the relationship between L'_{Aeq,8 h} and HFNIHL increased from 6.6% to 9.6% after the kurtosis adjustment. The DRR of L_{Aeq,8 h} and HFNIHL showed that the percentage of HFNIHL decreased after the adjustment of kurtosis in the non-steady noise groups, and the regression lines of the non-steady noise groups approached that of the steady noise group. The R² of the K₂ group increased from 0.935 3 to 0.986 3, and the R ² of the K₃ group increased from 0.905 6 to 0.951 6. Under the un-adjusted condition, the NIPTS₃₄₆ underestimation for the K₃ group was significantly higher than that for the steady noise group (t=−3.23, P=0.001). After the L_{Aeq,8 h} was adjusted by kurtosis, the NIPTS₃₄₆ underestimation decreased significantly in the three kurtosis groups (K₁: t=6.78, P＜0.001; K₂: t=14.31, P＜0.001; K₃: t=11.06, P＜0.001). There was no significant difference in the degree of underestimation between the three kurtosis groups (K₁ vs K₂: t=−0.22, P=0.830; K₁ vs K₃: t=−1.40, P=0.205) as the curves of the three kurtosis groups were nearly overlapped. Conclusion The kurtosis-adjusted L_{Aeq,8 h} can effectively estimate the hearing loss associated with non-steady state noise.

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths, characterized by highly hypoxic tumor microenvironment. Hypoxia-inducible factor-1α (HIF-1α) is a major regulator involved in cellular response to changes of oxygen levels, supporting the adaptation of tumor cells to hypoxia. Bruceine D (BD) is an isolated natural quassinoid with multiple anti-cancer effects. Here, we identified BD could significantly inhibit the HIF-1α expression and its subsequently mediated HCC cell metabolism. Using biophysical proteomics approaches, we identified inhibitor of β-catenin and T-cell factor (ICAT) as the functional target of BD. By targeting ICAT, BD disrupted the interaction of β-catenin and ICAT, and promoted β-catenin degradation, which in turn induced the decrease of HIF-1α expression. Furthermore, BD could inhibit HCC cells proliferation and tumor growth in vivo, and knockdown of ICAT substantially increased resistance to BD treatment in vitro. Our data highlight the potential of BD as a modulator of β-catenin/HIF-1α axis mediated HCC metabolism.