Objective To investigate the changes of regulatory T cells (Tregs) in hepatocellular carcinoma(HCC) patients with thermal ablation therapy(TAT),and to explore the mechanisms of immunity enhancement induced by TAT.Methods A total of 36 patients with HCC undergoing TAT (observation group) were enrolled in this study from December 2009 to December 2010.Another 20 healthy person were selected as control group.The percent of Tregs was detected by flow cytometry analysis.The level of serum IL-10 was measured with ELISA.Results Before treatment,the percent of Tregs in TAT group was significantly higher than that in control group [(11.44 ± 2.74)％ vs.(2.45 ± 0.67)％] (P ＜ 0.01).After treatment,the percent of Tregs in TAT group decreased significantly[(11.44 ± 2.74)％ vs.(5.07 ± 0.82)％] (P ＜ 0.01).The level of serum IL-10 in TAT group [(41.66 ± 9.24) mg/L] was significantly higher than that in control group [(14.32 ± 3.64) mg/L] (P ＜ 0.01).After treatment,the level of serum IL-10 in TAT group decreased significantly[(41.66 ± 9.24) mg/L vs.(25.47 ± 5.43) mg/L] (P ＜ 0.01).Conclusion TAT can enhance immunity in HCC patients,and which may accomplish by down regulation of Treg and IL-10.

A reusable chronocoulometric adenosine triphosphate (ATP)-aptamer sensor was developed in this work.A short chain of DNA marked as cDNA containing complementary sequence was immobilized on gold electrode based on Au-S self-assembly.The ATP aptamer was hybridized with cDNA.The surface-confined DNA could bind with [Ru(NH3)63+ (RuHex) in the electrolyte via electrostatic interaction.Upon target ATP binding, the aptamer confined onto electrode surface was disassociated from the cDNA oligonucleotides into the solution.Such surface density change of DNA lead to the decrease of chronocoulometric signal for the RuHex which confined on the electrode surface.The chronocoulometric signals showed a linear relationship with logrithm of ATP concentration in the range of 1 nmol/L to 100 μmol/L, and the detection limit of this aptamer sensor could reach 0.5 nmol/L (S/N=3).This aptamer sensor could be regenerated 5 times by simple steps.With this aptamer sensor, the basal level of ATP in the brain cortex micorodialysate was determined to be 19.2±3.7 nmol/L (n=3).

Objective To detect the prevalence of myositis-specific autoantibodies (MSAs) and myositis-associated autoantibodies (MAAs) in patients with polymyositis (PM) and dermatomyositis (DM), and analyse the correlation between MSAs and the clinical features and prognosis of PM/DM. Methods Serum samples of 31 PM and DM patients were screened for MSAs (including anti Jo-1, anti Mi-2, anti PL-7, anti PL-12 antibodies) and MAAs (including anti Ku, anti PM-Scl antibodies) by immunoblotting test. Results Serum MSAs/MAAs were detected in 18 out of 31 PM/DM patients (58%). MSAs were present in 12 patients (39%). The most frequently encountered MSAs was anti-Jo-1 autoantibody (29%), followed by anti-Mi-2 (7%), anti-PL-7 (3%), and anti-PL-12 (3%). MAAs were present in 10 patients (32%), including anti-Ku-72 (16%), anti-Ku-86 (23%) and anti-PM/Scl (7%). Notably, anti-Jo-1 antibody was closely associated with interstitial lung disease (ILD) and arthritis/arthralgia compared with the anti-Jo-1 antibody negative patients (P

Objective Duchenne muscular dystrophy(DMD)is one of the most common X-linked recessive neuromuscular degeneration diseases.It is caused by genetic defects of dystropin gene with deletion,duplication,or point mutation that results in clinical muscle fatigue and dystrophy.Usually,gene deletion of one or a few exons of dystrophin accounts for about 55%～65% patients,duplication for about 5%～10% patients and point mutation for 25%.Most of hot-spot deletion mutation of DMD can be detected by multiplex PCR and the point mutation can be detected by PCR/sequencing analysis,however,it remains a challenge to detect duplication.The recently developed MLPA(multiplex ligation-dependent probe amplification)is an efficient procedure that can accurately analyze the copy number and deletion mutation of whole dystropin gene.Methods A validation for simultaneous detection of entire dystropin gene was performed with two reactions.Both of which detect 39 and half exons of dystrophin gene.Results Nine out of 15 patients with DMD were found to have deletion mutation in different exons of dystrophin gene.Among these 9 patients,7 were found having deletion previously with multiplex PCR for mutation of hot-spot by Peking Union Medical University.Two patients who had not been found deletion by multiplex PCR were shown to have rare deletion at exon 18 or 43 in this study.Conclusions MLPA provides a simple,rapid and accurate method of simultaneously detecting homozygous,heterozygous deletions and duplication mutation in two single reactions for all exons of dystrophin gene,which may be applied into clinical molecular analysis for DMD.

Objective To determine the serum level of KL-6 in patients with polymyositis(PM)and dermatomyositis(DM),and to investigate the possible diagnostic value for interstitial lung diseases(ILD)in pa- tients with PM/DM.Methods Serum KL-6 concentrations were measured by ELISA in 53 adult PM/DM pa- tients,the control groups of 22 patients with infectious lung disease,and 50 healthy subjects.The association with clinical features and serum KL-6 levels was analyzed.Results The serum levels of KL-6 were(1543?761)、(429?106)、(336?196)and(289?105)U/ml in PM/DM patients with ILD and without ILD,patients with infectious lung disease and healthy controls,respectively.Serum KL-6 levels in the PM/DM patients with ILD were significantly higher compared with PM/DM patients without ILD,patients with infectious lung disease and healthy controls(all P＜0.01 ).However,no significant differences of serum KL-6 levels was found among PM/ DM patients without ILD,patients with infectious lung disease and healthy controls(P＞0.05).Significant cor- relations were found between the elevated levels of serum KL-6 and the presence of ILD in patients with PM/ DM(P＜0.01).The sensitivity and specificity of serum abnormal KL-6 levels for ILD in patients with PM/DM were 90.9% and 80.6%.Additionally,follow-up study showed the mean serum levels of KL-6 in six patients who died were significantly higher than that in other PM/DM patients with ILD(P＜0.01).Conclusion Serum KL-6 level is a reliable serum marker for ILD in idiopathic inflammatory myopathies,which may contribute to early differentiate ILD from lung infectious disease.Increased serum level of KL-6 may predict a poor out- come.

Objective To study the expression of major histocompatibility complex(MHC)on muscle biopsy specimens of idiopathic inflammatory myopathies(IIM),and assess diagnostic value of MHC in IIM. Methods Forty-five patients with IIM(19 polymyositis and 26 dermatomyositis)were selected for this study.Thirty healthy subjects were included as controls.Immunohistochemical staining was applied to identify the expression of HLA-A/B/C and HLA-DR on muscle biopsy-specimens in polymyositis (PM)/deFmatomyosiris (DM) patients and healthy controls.Results HIA-A/B/C antigens were expressed in muscle fibers in 18 out of 19 PM patients(95%),24 out of 26 DM patients(92%) and 3 out 0f 30 healthy controls(10%) respectively.The positive expression rate of HLA-DR in PM,DM patients and healthy controls were 84%, 81%,13%,respectively.The expressions of HLA-A/B/C and HLA-DR were significantly increased in PM and DM patients than those in healthy controls(both P<0.05),but no significant differences were found between PM and DM groups(both P>0.05).No significant correlations were demonstrated between the over-expression of HLA-A/B/C or HLA-DR and the extent of inflammatory infiltrations,muscle damage or clinical features in PM/DM groups (all P values>0.05).Conclusion MHC-Ⅰ and MHC-Ⅱover-expression in muscle fibers are the early events in PM and DM,and may occur in the absence of lymphocyte infihration and muscle damage.Immunostaining for MHC-Ⅰ and MHC-Ⅱcan be used as a routine test in the diagnosis of PM and DM.

Objective To study the shift handover experience of ICU clinical nurses. Method About 19 ICU nurses were enrolled in the investigation using semi-structured interviews and the results were analyzed with phenomenological analysis. Result Their shift handover experience were summarized into 6 themes, that is high recognition on the importance of shift handover, lack of standardized processes, incomplete contents of shift handover, frequent interruption, forgotten information and repeated information. Conclusions Shift handover is an important part of ICU nursing function but there are many problems in the practical operation. The nurse managers need to standard the handover process, stipulate handover contents and enhance the training on shift handover with specialist examination and positive results for the purpose of improving the quality of the shift handover.

Objective We reported previously that single nucleotide polymorphisms SNP) + 11G ＞ A in intron 3 of the human urate transporter 1 (hURAT1) gene are associated with hyperuncaemia in Han Chinese.The aim of the present study was to evaluate the effect of the variants on hURAT1 function.Methods The wild-type,mutant-type hURAT1 and exon 5-null hURAT1 were constructed,and respectively microinjected into the zebrafish embryo yolks.The subcellular localization of different genotypes of hURAT1 was detected by confocal laser scanning microscope.Results Compared with wild type,the mutant recombinant plasmid transcribed two types of mRNA spliceosome,the wild type and the exon 5-null type.The hURAT1 wild type protein was prominent localized on cell membrane,while the mutant type and exon 5-null hURAT1 proteins were distributed uniform in the cytoplasm but not on the cell membrane.Conclusion The hURAT1 variant + 11 G ＞ A resulted in an alternative splicing of hURAT1 mRNA-exon 5-null type.Its protein product exhibited a different subcellular localization compared with that of wild type.

Adult ; Ambroxol ; administration & dosage ; therapeutic use ; Expectorants ; administration & dosage ; therapeutic use ; Female ; Glycoproteins ; therapeutic use ; Humans ; Lung ; pathology ; Lung Injury ; drug therapy ; pathology ; Male ; Paraquat ; poisoning ; Respiratory Distress Syndrome, Adult ; drug therapy ; etiology ; Retrospective Studies ; Young Adult

AIM: To obtain differentially expressed cDNA fragments in the cerebellum of rats and screen unknown expressed sequence tag (EST). METHODS: Suppression subtractive hybridization (SSH) was carried out, in which the cDNA fragments of cerebellum were taken as "tester" and correspondingly that of cerebrum and brain stem as "driver". The homogeneous sequences between the tester and driver were excluded and the rare sequences in cerebellum were enriched by SSH. The differentially expressed cDNA fragments were further cloned for the construction of subtracted cDNA libraries and sequencing. RESULTS: 32 clones were selected and 34 cDNA fragments were sequenced. 8 of 32 were proved to be true positive with reverse Northern assay, 13 of 34 fragments were identified to be new cDNA fragment and given the gene sequence numbers by GenBank (AW288461-AW288474). CONCLUSION: SSH is very useful method for screening differentially expressed genes. Our data may be helpful to understand the molecular mechanism of brain function.