Objective To investigate the role of peripheral blood lymphocyte apoptosis through Fas/ Fas ligand pathway in the development of viral myocarditis. Methods Twenty-five viral myocarditis (VMC) and 25 healthy children were chosen for study. The apoptosis percentage in peripheral blood iymphocytes was quantitatively examined, flow cytometry with Annexin V/PI double parameter was used. And the expressions of Fas and Fasl protein in peripheral blood lymphocytes were analyzed with flow cytometry. Results The percentage of apoptosis in peripheral blood lymphocytes in children with VMC [(0. 6976 ± 0. 1109)%] was significantly higher than that of healthy children [(0. 1288 ± 0. 1069) %] (t = 18.459, P ＜ 0.001). Moreover, the apoptosis cell percentage showed a significant positive correlation with the serum levels of myocardium enzyme (creatine kinase-MB, lactate dehydrogenase, creatine kinase, hydroxybutyrate dehydrogenase)(r =0. 817 ,P ＜0. 01 ;r =0. 785 ,P ＜0.01 ;r =0. 726 ,P ＜0. 01 ;r =0. 712 ,P ＜0. 01). The expression of Fas and Fasl protein in VMC patients(2. 8804 ± 0. 3020,4. 4496 ± 0. 4836) were higher than those of health children(2. 1300 ±0. 6200,0. 1238 ±0. 0384) ,which had significant difference(t = 17. 321 ,P ＜0. 001 ;t' =44. 577, P ＜ 0. 001). The apoptosis percentage of peripheral blood lymphocytes in children with VMC showed a significant positive correlation with the Fas and Fasl protein expression in peripheral blood lymphocytes(r =0. 972, P ＜ 0. 01; r = 0. 958, P ＜ 0. 01). Conclusion Apoptosis percentage of peripheral blood lymphocytes increases among children with VMC, probably because immune response decreased or it is related with the degree of myocardium damaged. Fas and Fasl play an important role in apoptosis of peripheral blood lymphocytes in children with VMC.

Adult ; Female ; Humans ; Nasal Cavity ; Nasal Lavage ; Nasal Obstruction ; prevention & control ; Pregnancy ; Pregnancy Complications ; prevention & control ; Rhinitis ; prevention & control ; Saline Solution, Hypertonic ; administration & dosage ; therapeutic use ; Young Adult

Child, Preschool ; Coronary Aneurysm ; etiology ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; complications ; pathology

Objective To investigate the relationship between pure -tone audiometry (PTA ) ,speech dis‐crimination abilities in quiet or in noise and the international outcome inventory for hearing aids (IOI-HA) in pres‐bycusis patients .Methods Twenty presbycusis subjects were tested in this study .Pure-tone audiometry (PTA) and speech discrimination threshold were obtained before being fitted with hearing aids .They weared hearig aids more than six months ,and pure-tone audiometry ,speech discrimination scores in quite(the level = 65 dB SPL) and in noise(signal to noise ratio = 10 dB) were carried out in sound field .A stepwise forward multiple-regression analysis was performed to investigate the impact of PTA and speech discrimination scores to IOI-HA .Results The PTAs before or after hearing aid fittings showed a negative association with IOI-HA ,while speech discrimination scores in quiet or in noise before or after hearing aid fittings showed a positive association with IOI-HA .Speech discrimination threshold in noise was identified as a single predictor of IOI-HA(P<0 .001) .Conclusion The relation between speech discrimination scores in noise and IOI-HA suggests that a poor score might limit the hearing aids outcome .The speech discrimination scores in noise help the clinicians predict the outcomes of hearing aid in real‐ities .

Objective To construct eukaryotic expression vector of wild type E 4F1 and the mutant deleting amino acid region 32-81, and to detect the interaction between wild type or mutant E 4F1 and p53 and to study the effect of E4F1 on the expression level of p21.Methods Wild type and mutant sequences of E 4F1 were amplified from the mammary library using standard PCR and recombinant PCR .The sequences were cloned into pXJ 40-MYC vector to generate the MYC-E4F1 and MYC-E4F1(Δ32-81) recombinant plasmids that were transfected into 293T cells and identified by Western blotting . FLAG-p53 and MYC-E4F1 or MYC-E4F1(Δ32-81) were co-transfected into 293T cells and immunoprecipitation assay was performed to detect the interaction of wild type or mutant E 4F1 with p53.Wild type and mutant E4F1 expressing vec-tors were co-transfected into osteosarcoma U2OS cells and the expression of p21was detected.Results Recombinant plas-mids of MYC-E4F1 and MYC-E4F1(Δ32-81) were successfully constructed.Both wild type and mutant E4F1 interacted with p53.Deletion of amino acid region 32-81 of E4F1 increased the interaction .The expression level of p21 was in-creased by wild-type E4F1, but not by mutant E4F1.Conclusion The eukaryotic expression vector of wild type E4F1 and its deletion mutant is successfully constructed .Both of them interact with p53.Deletion of amino acid region 32-81 of E4F1 increases the interaction .This study contributes to further studies on the regulation and mechanism of E 4F1 on p53.

AIMTo develop a near-infrared diffuse reflectance analysis (NIRDRA) method for rapid noninvasive quantitative determination of ambroxol hydrochloride in half-finished product particles and non-blister-packed, blistered tablets.

METHODSAll spectra were measured with a Fourier transform spectrometer equipped with a PbS and a InGaAs detector, an external integrating sphere, a rotating sample cup, and a fibre-optic probe for reflectance measurements. All samples were scanned from 12,000 cm-1 to 4,000 cm-1, and each sample spectrum was obtained as an automatic mean of 64 scans. No spectrum pre-processing method was used, and spectral regions, 4,602-4,247, 12,000-7,498 and 6,102-5,446, 12,000-5,446 cm-1 were selected to develope mathematical models by partial least square method for half-finished product particles and non-blister-packed, blistered tablets samples, respectively.

RESULTSThe optimal rank and mean square error determined for half-finished product particles and non-blister-packed, blistered tablets samples by cross validation method all was 6 and 0.306, 0.972 and 1.492, respectively, the average recovery was 100%, 100% and 102% respectively; and the RSD was 1.17%, 1.70% and 1.78% respectively.

CONCLUSIONResults showed that the NIRDRA method was rapid, simple, noninvasive and sensitive, and it can be applied to assay the content of ambroxol hydrochloride in half-finished product particles non-blister-packed and blistered tablets.

Ambroxol ; administration & dosage ; analysis ; Expectorants ; administration & dosage ; analysis ; Quality Control ; Spectroscopy, Near-Infrared ; methods ; Tablets

Objective To evaluate the speech recognition ability in noise enviroment in older adults with age-related hearing loss and to discuss the influences of age ,hearing loss and cognitive function on the ability of speech recognition in noise enviroment .Methods A total of 90 older adults aged over 60 years at the hearing clinic of Xinhua hospital were divided into three groups according to the age ,which were 60~69 y group (88 ears) ,70~79 y group (64 ears) ,> 79 y group (28 ears) .All the participants completed pure tone audiometry ,the Mandarin hearing in noise test (MHINT) and the mini-mental state examination (MMSE) .The pure tone average hearing threshold (PTA) ,signal-to -noise ratio (SNR) of MHINT and MMSE scores were obtained .We used the statis_tical software SPSS 17 .0 to analyze the correlations among age ,PTA ,adaptive SNR and MMSE scores .ResuIts ①PTA and SNR increased with the age .The difference of PTA and SNR for different age groups was statistically sig_nificant (P<0 .01) .②The difference SNR among different PTA groups were significant (P<0 .01) ,and when the influence of MMSE scores was controlled the difference was still significant .There was a significant linear correlation between PTA and SNR (r=0 .326 ,P<0 .01) .MMSE scores decreased with the increasing of PTA ,and there was also a linear correlation between PTA and MMSE scores (-0 .187 ,P<0 .05) .③Adaptive SNR and MMSE scores were correlated (r= -0 .201 ,P<0 .01) ,but there was no significant linear relationship .Besides ,SNR of the cog_nitive function impairment group (<27 points) was significantly greater than that of the normal cognitive function group (27~30 points) (P<0 .01) .However ,among the three PTA groups ,the differences were not statistically significant .ConcIusion The hearing in noise test reflects the ability of speech recognition in noise enviroment in ol_der adults ,and can be used to assess the hearing functions in elder .Speech recognition in noise ,peripheral hearing loss and cognitive functions are relevant .Cognitive functions have an influence on the evaluation of the hearing func_tion for older adults .

Objective To investigate the reliability and practicality of Chinese version the hearing handicap in-ventory for the elderly -screening version(HHIE -S) scale for elderly hearing screening in China .Methods We compared the HHIE -S scores with results of pure -tone audiometry test .A total of 840 Chinese individuals older than 60 years filled in the scale all by themselves first ,then refilled it according to explanations of each question translated by researchers .All participants finished pure -tone audiometry test with frequencies of 0 .25 ,0 .5 ,1 .0 , 2 .0 ,and 4 .0 kHz .Sensitivity ,specificity ,positive predictive value (PPV ) ,false-negative rate and false -positive rate were compared with HHIE -S cut -off points of >8 or >10 and pure-tone averages (PTAs) greater than 25 ,40 and 60 dB HL .Analysis the area under the curve (AUC) values of receiver operating characteristic (ROC) curves for each three PTA levels .Results Cronbach's α of the scale was 0 .85 .HHIE -S scale pre -explanatory score was 7 .6 ± 10 .6 ,and post-explanatory score was 10 .8 ± 16 .7(P<0 .01) .HHIE -S> 8 points higher than H HIE-S> 10 points in the sensitivity ,positive predictive value increase ,specificity ,false-positive rate ,false negative rate decreased ,paired samples t - test sensitivity and false -negative rate (P< 0 .05) .After the guide meanwhile HHIE-S cut-off points of > 8 when grouping with PTA> 25 dB HL ,PTA> 40 dB HL and PTA>60 dB HL ,conducted ROC curve analysis ,the AUC value was 0 .70 ± 0 .03 ,0 .84 ± 0 .01 and 0 .88 ± 0 .02 .Conclusion The score of HHIE-S scale>8 points can be used when PTA> 40 dB HL as goal for hearing loss of old people , it can be more accurate after explanations ,so HHIE-S scale can be used as a preliminary hearing screening method for Chinese elderly people .

OBJECTIVETo explore the possibility of cell-medicated immune response induced with heat shock protein 70 (HSP70)-HBsAg protein complex in vitro.

METHODSHSP70-HBsAg complex was reconstituted in vitro which was injected into mice in order to observe that whether HSP70-HBsAg would stimulate humoral and cellular immune responses. HSP70, HSP70-HBsAg complex and HBsAg were used to activate the dentritic cell (DC) individually, which would initiate homogeneic T lymphocyte to transform to cytotoxic T lymphocyte (CTL). The cytotoxicity of CTL was detected with MTT assay.

RESULTSHSP70-HBsAg complex elicited both humoral and cellular immune responses against HBsAg in mice. Specific CD8+ CTL response was readily induced by HSP70-HBsAg complex and HBsAg, especially the former.

CONCLUSIONSHSP70-HBsAg complex is immunogenic and HSP70 can lead to great efficient CTL response. And HSP70-HBsAg complex may be used as a protein vaccine for immunotherapy for chronic hepatitis B.

Animals ; Cells, Cultured ; Dendritic Cells ; immunology ; secretion ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; chemistry ; Peptides ; chemistry ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo study the expression of the shortened hepatitis B surface antigen in prokaryocyte and detect the antigenic characters.

METHODSFirstly, the gene fragments coding the 152 and 124 amino acids of the carboxyl terminus of hepatitis B surface antigen (HBsAg) were amplified by polymerase chain reaction (PCR). Secondly, they were cloned to plasmid pBKS+, and the accuracy of those constructions were confirmed by restriction enzyme digestion and DNA sequencing. Then, they were cloned to prokaryocytic expression vector-plasmid pET32a(+). The recombinant plasmids were transfected into E.coli BL21 and induced to express with IPTG.

RESULTSThe recombinant plasmids were successfully constructed. In E.coli BL21, the protein was expressed in a fusion fashion and could be recognized by monoclonal antibody against HBsAg with ELISA and Western blot.

CONCLUSIONThe shortened HBsAg can be expressed in prokaryocyte.

Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B virus ; genetics ; Humans ; Plasmids ; Polymerase Chain Reaction ; Prokaryotic Cells ; metabolism ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology