Objective Through exploring the concordance of objective multi-parameters analysis and perceptual evaluation,to establish an objective multi-parameters evaluation protocol of voice disorder and to make the evaluation of voice objectification and quantification. Methods Voice samples from 271 patients ( 124 female and 147 male)with dysphonia and 69 control subjects with normal voice (37 female and 32 male)were recorded and assessed by a jury composed of 5 experts in phoniatrics from different hospitals.The jury was instructed to classify voice samples according to the G (grade) component of the GRBAS scale on a visual analogue scale secondarily transformed in a 4-point scale ranging from 0 for normal to 3 for severe dysphonia. The voice samples were unified sentences and ordered randomly 3 times,the mean of 3 evaluation scores were the final results.The objective parameters,including fundamental frequency ( F0),jitter,shimmer,fundamental frequency standard deviation (FOSD),normalized noise energy (NNE),harmonic-to-noise ratio (HNR) and maximal phonatory time ( MPT),were measured on a 2-second sustained vowel/a/ including its initial segment,using the software Dr.Speech for Windows.The data were analyzed using SPSS11.0.Results All objective parameters except for F0 had high correlation with G and the variance tendency of these parameters values was coherent with the extent of voice disorder.And there were statistical differences between adjacent voice disorder groups. Male and female objective multi-parameters protocols were established respectively consisting of jitter,shimmer,FOSD, NNE,HNR and MPT using discriminant analysis ( P ＜ 0.05 ). The concordance between perceptual evaluation and objective multiparameters evaluation was 81.6％ in male and 83.2％ in female.The concordance of evaluation of normal voice and severe voice disorder groups were better than that of mild and moderate voice disorder groups.All mis-grading voices were judged in the adjacent voice group.Conclusions The objective parameters of voice are able to reflect the characteristic of its perceptual evaluation and the concordance between perceptual evaluation and objective multi-parameters evaluation is good. The objective multi-parameters evaluation protocol we established could provide an objective and quantitative evaluation method for voice disorders.

ObjectiveTo explore the expression of CD3+ CD8+ human leukocyte antigen (HLA)-A2+T lymphocytes with specificity to the different hepatitis B virus (HBV) peptides in the peripheral blood mononuclear cells (PBMC)from the patients with hepatitis B associated hepatocellular carcinoma (HCC).MethodsThe HLA-A2+ PBMC from four patients with hepatitis B associated HCC were incubated with five HBV/HLA-A2 pentamers respectively,which were HBV sAg (FLLTRILTI),HBV sAg (GLSPTVWLSV),HBV sAg (WLSLLVPFV),HBV core (FLPSDFFPSV),and HBV pol (FLLSLGIHL),as well as anti-CD3-pacific blue and anti-CD8-fluorescein isothiocyanate (FITC).Then,HBV/HLA-A2-CD3-CD8 positive cells were detected by flow cytometry. The monoclonal HBV/HLA-A2-CD3-CD8+ cells were acquired by fluorescenceactivated cell sorter,and cultured and identified by flow cytometry.The anti-HBV specific T lymphocytes were then cultured with HepG2 (HLA-A2+ ) cells and the release of interferon γ (IFN-γ)were determined by enzyme-linked immunosorbent assay (ELISA),Res(a)ltsThe percentage of antiHBV T lymphoeytes with specificity to GLSPTVWLSV in total CD8+ T lymphoeytes from four patients with hepatitis B associated HCC was 1.44％±0.04％,which was higher than those to other four HBV antigen peptides (0.68％±0.08％ of FLLTRILTI,1.06％±0.09％ of FLPSDFFPSV,0.56％ ±0.04％ of FLLSLGIHL,and 0.46％ ±0.08％ of WLSLLVPFV) (t=0.001,P＜0.05).The two lines of monoclonal cell with specificity to GLSPTVWLSV both exhibited high level of IFN-γ expression after incubated with hepatic carcinoma cell line HepG2 (HLA-A2+)with HBV GLSPTVWLSV peptide.ConclusionsCD3+ CD8+ HLA-A2+ cells with specificity to the different HBV peptides exist in PBMC of patients with hepatitis B associated HCC.The expression level depends on HBV antigen peptide sequences and genomic sites.

Objective To learn the stroke occurrence status and the influencing factors of the patients with type 2 diabetes,and to provide the basis for the prevention of stroke.Methods Patients with type 2 diabetes which were reported in 2011 year from the system of chronic disease monitoring information management of Zhejiang Province were selected and followed -up until October in 2014.Stroke occurrence were recorded and the influencing factors were analyzed.Results The incidence rate was 5.22％ (59/1 129),and the annual incidence was 1.56％.Fifty six cases (94.92％) were ischemic stroke.Logistic regression analysis showed that older than 65 and hypertension were risk factors for type 2 diabetes complicated with stroke.The group of 65-and 75-were associated with higher risk of stroke than 45-(OR =3.38,95％ CI:1.39-8.24;OR =7.77,2.14-28.24).Hypertensionin were the influencing factors of stroke among patients with type 2 diabetes (OR =10.92,95％ CI:5.94-20.09).Conclusion Older age and hypertension were risk factors for type 2 diabetes complicated with stroke.Therefore effective control blood pressure among older patients with type 2 diabetes should be strengthed to prevent stroke.

Adolescent ; Adult ; Aged ; Child ; Colles' Fracture ; physiopathology ; therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Supination ; Treatment Outcome ; Young Adult

OBJECTIVETo study the effects of benzo(a)pyrene (BaP) on the cell cycle distribution and activities of mitogen-activated protein kinase (MAPK) signal molecules (ERK1/2, JNK1/2 and p38) in human embryo lung cells (HELF), and to investigate the relationship between alterations of MAPK protein phosphorylation and the cell cycle distributions.

METHODSThe phosphorylation of MAPK were induced by exposing HELF cells to BaP at 0.1, 0.5, 2.5 and 12.5 micromol/L. The phosphorylation and protein expression levels of ERK1/2, JNK1/2 and p38 were determined through western-blotting assay. And the flow cytometry assay was used to measure the cell cycle effects in HELF cells after treatment with 2.5 micromol/L BaP for 24 h.

RESULTSThe phosphorylation levels of ERK1/2, JNK1/2 and p38 were significantly increased through BaP exposure. In addition, the phosphorylation of these three MAPKs has similar alteration pattern. We found that exposure of cells to 2.5 microM of BaP for 24 h resulted in a decrease of G(0) and G(1) population by 11.9% (F = 41.38, P < 0.01) and an increase of S population by 17.2% (F = 68.13, P < 0.01). Three chemical inhibitors of MAPK (AG126, SP600125 and SB203580) could significantly inhibit the cell cycle alteration because of BaP treatment.

CONCLUSIONERK1/2, JNK1/2 and p38 could positively regulate the BaP independently induced cell cycle alterations.

Benzo(a)pyrene ; toxicity ; Cell Cycle ; drug effects ; Cells, Cultured ; Fibroblasts ; drug effects ; metabolism ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Lung ; cytology ; embryology ; MAP Kinase Kinase 4 ; metabolism ; MAP Kinase Signaling System ; drug effects ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinase 8 ; metabolism ; Mitogen-Activated Protein Kinase 9 ; metabolism ; Signal Transduction ; drug effects ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo evaluate the efficacy of mitoxantrone combined high dose of cytarabine and recombinant human granulocyte colony-stimulating factor (MAG) regimen for mobilizing autologous peripheral blood stem cells (APBSC) in patients with hematopoietic malignancies.

METHODSFrom December 1995 to April 2003, 14 lymphoma and 29 acute leukemia patients were treated with high-dose cytarabine (2 g/m2 every 12 h, days 1 and 2) and mitoxantrone (10 mg/m2, days 2 and 3), followed by 300 microgram recombinant human granulocyte colony-stimulating factor per day (rhG-CSF 300 microg/d) i.e, the MAG regimen as mobilization regimen of peripheral blood stem cells. rhG-CSF was given subcutaneously when the white blood cell (WBC) count below 1.0 x 10(9)/L following the MA chemotherapy, APBSC were harvested when WBC count increased using Baxter CS3000plus or Cobe Spectra.

RESULTSMobilization was successful in 13 of 14 lymphoma patients with MNC (3.91 +/- 2.70) x 10(8)/kg, CD34+ cells (17.79 +/- 12.90) x 10(6)/kg. Meanwhile, mobilization was successful in 24 of 29 acute leukemia patients with average of 2.13 times for apheresis. The median MNC and CD34+ cells yielded were 3.62 x 10(8)/kg and 7.37 x 10(6)/kg respectively, rhG-CSF was used for a median time of 7 days. Excepting for grade I-II gastrointestinal toxicity in 8 and infection in 14 cases, no major side effects were observed. There was no mobilization-related mortality. Minimal residual diseases became undetectable after mobilization in some patients.

CONCLUSIONMAG is a safe and highly effective mobilization regimen in patients with lymphoma and acute leukemia.

Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cytarabine ; administration & dosage ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; drug effects ; Humans ; Lymphoma ; therapy ; Male ; Middle Aged ; Mitoxantrone ; administration & dosage ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; therapy ; Recombinant Proteins

OBJECTIVETo establish multiple short tandem repeat (STR) amplification by fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis for quantitative determination of chimerism, and to evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThirty-one patients received bone marrow transplantation (BMT) or nonmyeloablative allogeneic stem cell transplantation (NST) were evaluated. Peripheral blood and bone marrow were co-llected before and after transplantation in different period. Nine different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus PCR amplification kit. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype software were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient.

RESULTS48.4% of the patients received sex-matched transplantation and the quantification of donor chimerism could only be performed by STR-PCR method. Comparison of values obtained by FISH analysis with that by STR-PCR in patients transplanted from sex-mismatched donors showed an excellent correlation. The median number of informative alleles was 6.7 (range 2 - 10). The donor's alleles appeared in all the patients on day 7 post-transplant. The median values of donor chimerism in BMT group were inferior to that in NST group on day 7, day 14 and 1 month post-transplant. However the difference disappeared in the midterm or later period of transplant. On day 21, all of the 31 patients had stable engraftment and the percentage of donor chimerism was more than 92%. Median follow-up was 17 (3.5 - 29.0) months after transplantation. Twenty-six of 31 patients had durable engraftment and donor chimerism ratio was more than 90%. So for all of them survived leukemia-freely. Four of the 31 patients had unstable mixed chimerism and relapsed within 6 months post allo-HSCT. Another patient with unstable mixed chimerism appeared graft rejection. Decreasing values of donor chimerism were detected prior to the occurrence of graft rejection and disease relapse. The incidence of GVHD was much higher in the group of full donor chimerism.

CONCLUSIONSequential and quantitative monitoring of STR is a valuable tool for studying engraftment dynamics, graft rejection, and relapse and for predicting GVHD. Furthermore it can provide a basis for early intervention of clinical treatment.

Adolescent ; Adult ; Child ; Electrophoresis, Capillary ; Female ; Graft Rejection ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Polymerase Chain Reaction ; Recurrence ; Tandem Repeat Sequences ; Transplantation Chimera ; Transplantation, Homologous

OBJECTIVETo explore the impact of IL-2- and IL-15-activated donor natural killer (NK) cell infusion on graft-versus-host-disease (GVHD) and graft-versus-leukemia (GVL) effect post allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThe C57BL/6 mice splenic NK cells were selected by microbeads, and then expanded in the media containing IL-2 and IL-15. The killing activity of NK cells was detected. In the leukemia mouse model, recipients (BALB/c) were intravenously inoculated with EL9611 leukemia cells 8 days before transplantation. Lethally irradiated BALB/c recipient mice were transplanted with 5 x 10(6) bone marrow cells (BMCs), or 5 x 10(6) BMCs plus 1 x 10(7) splenocytes with or without 1 x 10(7) activated NK cells. Additionally, NK cell infusion group mice were intraperitoneally injected with a mixture of IL-2 and IL-15 post transplant. Survival time, GVHD occurrence, lineage chimerism, TRBV spectra-typing were observed post transplant.

RESULTSThe purity of isolated splenic NK cells was 95.7% - 97.1%. The killing activity of NK cells after activation was increased by 3 times. GVHD did not occurred in allogeneic BMCs infusion group, whereas did from 1 week after transplant in allogeneic BMCs + splenocytes infusion group. The severity of GVHD in total body irradiation (TBI) experimental group was significantly lower than in splenocytes infusion group (P < 0.05). The survival time was 9.5 - 14.0 d in TBI alone conditioning group. In leukemia mouse model, 100 day survival rate was 10% the rest of them were died of leukemia while in experimental group, the more than 100 days survival rate was 80% (P < 0.01). PB NK cells at 2 week post-transplant were 4.8% in experimental group and 2.8% in control group. NK cells recovery in experimental group was earlier than that in control group (P < 0.05). TRBV reconstitution was faster in experimental group than in control group, moreover, the number of TRBV family expression was more in experimental group than in control group which mainly expressed monoclone or oligo-clone.

CONCLUSIONSDonor alloreactive NK cells can be efficiently expanded and activated with IL-2 and IL-15. Donor activated NK cell infusion and IL-2, IL-15 treatment can promote immune reconstitution, mitigate GVHD and reduce leukemia relapse.

Animals ; Cells, Cultured ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; Hematopoietic Stem Cell Transplantation ; Interleukin-15 ; immunology ; pharmacology ; Interleukin-2 ; immunology ; pharmacology ; Killer Cells, Natural ; cytology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL

To explore the absorption mechanism of paeonol-beta-CD from various intestinal segments and offer biopharmaceutics data for paeonol new dosage form. The absorption kinetics and permeability rate consatants were investigated by the in situ perfusing method in rats. The absorption of the drug conforms to the firt-order kinetics and passive transport mechanism . The results indicate that paeonol-beta-CD absorption mechanism wasn't change.
Acetophenones ; pharmacokinetics ; Animals ; Intestinal Absorption ; physiology ; Intestines ; metabolism ; Kinetics ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVEIt was noticed that coxsackievirus A16 (CA16) and enterovirus 71 (EV71) were two major etiological agents of hand, foot and mouth disease (HFMD) in children. Recently there were several large outbreaks of HFMD in the Asia-Pacific region, and there was a propensity to cause severe complications or death in children under 5 years of age. The severe forms were associated with EV71 infection. Although epidemics of HFMD have been reported in the mainland of China, few reports about EV71 as the pathogen of HFMD epidemics are available. The present study was conducted to investigate the causal agent of an HFMD epidemic in children in Shanghai from April to June of 2002.

METHODSTotally 102 specimens (including vesicle fluid, stool and throat swabs) were collected from 72 patients with HFMD. The specimens were inoculated into Vero and/or RD cells. At first all the isolates were respectively neutralized by the RIVM pools of enterovirus antiserum, the type-specific antisera to EV71 or to CA16. Secondly all untyped isolates were tested by RT-PCR assay with two specific primer pairs for VP1 genes of EV71 and CA16 respectively. The EV71 and CA16 were identified depending on the size of PCR products. Sequence analyses of VP1 genes of 9 virus strains were performed by the laboratory of China CDC.

RESULTSViruses were isolated from 91 specimens from 67 patients. Serotyping by neutralization failed for all the isolates. But the RT-PCR results indicated that the viruses isolated from 78 specimens from 58 patients were identified as positive for CA16 and the isolates from 13 specimens from 9 patients were identified as positive for EV71, the ratio between CA16 and EV71 was 6.4:1. The results of sequence analyses were consistent with those of PCR assay. Two EV71 strains isolated in this study belonged to a new lineage (C4) within genogroup C. One patient with EV71-associated HFMD had a complication of encephalitis with convulsion, shock, coma and dyspnea.

CONCLUSIONCA16 and EV71 were the primary causes of HFMD during the epidemic. It was the first report of EV71-associated severe encephalitis occurred in patients with HFMD in Shanghai.

Animals ; Cercopithecus aethiops ; Child ; Child, Preschool ; China ; epidemiology ; Coxsackievirus Infections ; diagnosis ; epidemiology ; Disease Outbreaks ; Enterovirus ; isolation & purification ; Enterovirus A, Human ; isolation & purification ; Female ; Genes, Viral ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Male ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Vero Cells