Objective To evaluate if the assay of platelet-associated immunoglobulin G (PAIgG) can improve the sensitivity of monoclonal antibody immobilization of platelet antigens assay (MAIPA) in immune thrombocytopenic purpura (ITP).Methods Anti-GP Ⅱb/Ⅲa and anti-GP Ⅰb/Ⅸ autoantibodies were detected by a modified MAIPA;PAIgG was detected by competitive ELISA.MAIPA and PAIgG were done simultaneously in 190 patients with thrombocytopenia,in which 132 were diagnosed as ITP according to the diagnostic criteria Kappa test was used to analyze the concordance between MAIPA and PAIgG.Results The sensitivities of PAIgG alone or in parallel with MAIPA to ITP were 32.6% and 37.2% higher than MAIPA alone respectively,while their specifieities were 51.7% and 56.9% lower respectively.The sensitivity,specificity,positive predictive value,negative predictive value of PAIgG in series with MAIPA were all not superior to MAIPA alone.The kappa values between MAIPA and PAIgG in ITP and non-immune thrombocytopenia were 0.129 and 0.012 respectively,which meant the concordance between them was poor.Conclusion The detection of PAIgG should not be used as screening before MAIPA in distinguishing immune from non-immune thrombocytopenia.

Objective To study the clinical characteristics and outcome of cytomegalovirus(CMV) infection in infants.Methods All data including time of infection,clinical characteristics,laboratory tests and outcome of CMV infections in hospitalized infants were collec-(ted) and analyzed from January,1994 to July,2004.Results In 87 infected infants,congenitally infected newborns,perinatal infection in infants and postnatal infection in infants accounted for 27.6%,62.0%,16.6%,respectively.CMV hepatitis was the most frequent type of disease with the incidence of 41.3%,in which the incidence of splenomegaly was 10.3%.Most of CMV hepatitis infants had a good prognosis with the improved rate 80.5%.Central nervous system abnormality(including abnormal intension of muscle,convulsion,ocular and hearing abnormalities) occurred only in congenital and perinatal infection with the incidence of 20.4%.Generalized infection,the incidence of congenital infection and perinatal infection was 16.7%,1.8%,respectively.It did not occur in postnatal infection.The mortality rate of congenital infection and perinatal infection were 12.5% and 1.85%,respectively.Conclusions CMV infection is the main cause of infant hepatitis and it can also cause neurologic sequelae.The outcome of generalized infection in congenital infection is bad and the mortality rate is high.

OBJECTIVETo investigate the malondialdehyde (MDA) level and paraoxonase-1 (PON-1) activity in the serum and seminal plasma of infertile men with chronic viral hepatitis and their influence on the semen parameters of the patients.

METHODSWe collected serum and semen samples from 42 infertile men, 45 infertile males with chronic viral hepatitis, and 50 healthy fertile men as controls. We measured the MDA level in the serum and seminal plasma by spectrophotometry, detected the PON-1 activity by spectrophotometry, and determined the sperm DNA fragmentation index (DFI) by acridine orange fluorescence staining.

RESULTSThe MDA level was significantly higher but the PON-1 activity remarkably lower in the serum and seminal plasma of the infertile males with chronic viral hepatitis than in the healthy controls and infertile patients (P <0.01 or P <0.05). Total sperm motility and sperm survival rate were significantly lower while the sperm DFI markedly higher in the former than in the latter two groups (P <0.01 or P <0.05). No statistically significant difference was found among the three groups in sperm concentration (P >0.05). The WBC counts in the semen of the infertile and infertile with chronic viral hepatitis groups were significantly higher than that in the health controls (P <0.05). The MDA level and PON-1 activity in the seminal plasma were positively correlated with those in the serum in the infertile males with chronic viral hepatitis (r=0.57 or 0.48, P <0.01).

CONCLUSIONVirus-induced chronic active hepatitis enhances oxidative stress in the reproductive system, aggravates sperm damage, and affects sperm quality parameters.

Adult ; Aryldialkylphosphatase ; analysis ; Case-Control Studies ; DNA Fragmentation ; Fertility ; Hepatitis, Viral, Human ; complications ; Humans ; Infertility, Male ; blood ; Male ; Malondialdehyde ; analysis ; blood ; Oxidative Stress ; Semen ; Sperm Count ; Sperm Motility ; Spermatozoa

Objective: This study aims to investigate the positive practice environments evaluation of doctors in public hospital and provide suggestions for Chinese health care reform from the perspective of health service providers.Methods: Using self-designed questionnaire, a national cross-sectional survey was performed in 77 public hospitals across seven provinces/ metropolis, involving a total of 3564 doctors.Positive practice environment includes organizational management and doctor-patient two-dimension relationships.Results: The doctor''s mean score of positive practice environment evaluation was 18.02±4.86 (out of 40).Doctors had a higher evaluation (score) of organizational management than doctor-patient relationship.In the organizational management dimension, the personal advice got the lowest scores(2.24±1.04);and in doctor-patient relations, evasion of medical risk got the lowest scores(1.59±0.81).There was a difference in the positive practice scores of physicians in different regions, and the scores of doctors from the western China were the lowest (P<0.001.Doctors from western medicine hospitals had lower evaluation of positive practice environments than ones from Chinese Traditional Medicine hospitals(P=0.002.Conclusion: As per the analysis conducted in this investigation, the current medical practice environment of doctors in China is not a promising one.Doctor-patient relationship is worse than organizational management.Hospitals should adopt a participatory decision-making model in management and establish a broadband performance based compensation to motivate junior doctors.Besides, government should establish reasonable compensation incentive mechanism, with the aim of promoting doctor-patient trust.

The intracellular hepatitis B surface antigen (HBsAg) content per cell was increased by 7.2-fold in the culture with 1.5% dimethyl sulfoxide (DMSO) compared with that in the control without DMSO, while the extracellular HBsAg production and specific productivity were only improved by 70% and 3.2-fold, respectively. Electron microscope has been employed to reveal large dilated structures within recombinant CHO cells in the presence DMSO. The dilated structures have a distribution within whole cytoplasm, and some dilated areas were engulfed in the nucleus. These large, dilated structures were not observed in the control. Immunogold labeling was used to discover the accumulated HBsAg was localized within these dilated areas, and some HBsAg-specific labels were detected in the nucleus membrane, owing to the encroachment of the dilated areas upon nucleus. The result could help to reveal the mechanism of intracellular HBsAg accumulation in the presence of DMSO.

Objective: To screen candidate antigen for therapeutic HBV vaccine with a novel adjuvant DC-Chol. Methods: BALB/c mice were injected with DC-Chol liposome and HBsAg prepared from CHO and Yeast respectively. One week later, IL-4, IL-2, IFN-?were measured by ELISA or ELISPOT. Results: The levels of IL-2, IFN-?of HBsAg from Yeast with DC-Chol liposome were 20 and 119 times higher respectively than those of HBsAg from CHO with DC-Chol liposome. ELISPOT assay showed that the counts of spot-forming cells of IL-4 and IFN-?of HBsAg from Yeast with DC-Chol liposome were 2.8 and 46.3 times higher respectively than those of HBsAg from Yeast with Al(OH)3. Conclusion: HBsAg prepared from Yeast together with DC-Chol liposome may be an appropriate candidate for therapeutic HBV vaccine .

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Objective To explore the qualification and consuming of iodized salt at wholesale and household levels after Salt Iodization.Methods Iodized salt surveillance at wholesale and household levels every year by detecting iodine content.Direct titration method(GB/T 13025.7-1999)was used for salt iodjne detecting and arbitration method was used for Sichuan salt and special salt.Results Five thousand six hundred and seventy five samples of 227 batches from 3 wholesale industries were detected during 1996-2000,batch qualification rate was 60.79%(138/227)and iodized salt qualification rate was 61.83%(3509/5675).During 2001-2007,2556 samples of 252 batches from wholesale levels were detected.The batch qualification rate and iodized salt qualification rate were 1 00%(252/252)and 99.88%(2553/2556),respectively.At household level.1583 samples from 236 villages were detected during 1996-2000.Iodized salt qualification rate was 74.24%(1 170/1576)and consuming rate of qualified iodized salt was 73.91%(1 170/1583)and iodine median was 45.14 mg/kg.During 2001-2007,13 140 samples from 1656 villages were detected.Iodized salt qualification rate,consuming rate 0f qualified iodized salt and iodine median were 98.03%(12 830/13 088),97.64%(12 830/13 140)and 30.13 mg/kg,respectively. The most difference of iodine content was 3.46 mg/kg in 3 wholesale industries.At household level there was a 4.95%reduction in comparison with at wholesale level.Conclusions Salt iodization level and edible iodine salt reach the national requirements of iodine deficiency control from the starting stage.The quality 0f iodized saIt at household level related to the exclusive wholesale industry and loss phenomenon maybe existed when salt was sold from wholesale industries to residents.

OBJECTIVETo observe the localization of p53(301-393)(residues 301-393) in p53 positive/negative cells and its effect on cell mitosis.

METHODSThe protein expression of p53-GFP and p53(301-393)-GFP was checked by immunoblotting after transfection. Immunofluorescence staining was performed to detect the localization of wide type and mutant in Hela cells (p53 positive) and H1299 cells (p53 negative). The cell morphology of H1299 cells transfected of p53(301-393)-GFP and the cells in mitotic phase were observed. Cell cycle was analyzed by flow cytometry and p53 protein level in HeLa cells was evaluated by Western blot after transfection of p53-GFP and p53(301-393)-GFP.

RESULTSThe protein expression of p53-GFP and p53(301-393)-GFP was verified, p53-GFP was about 90 kMr and p53(301-393)-GFP about 40 kMr. Immunofluorescence microscopy demonstrated that both proteins were diffusely located in the nuclei of HeLa cells and H1299 cells. But different from the p53-GFP, the p53(301-393)-GFP was distributed in the nucleolus of HeLa cells. After transfection of the two plasmids, mitosis was inhibited in H1299 cells and some cells underwent apoptosis. G2/M progression of HeLa cells could be blocked by transfection of p53(301-393)-GFP, but endogenous p53 protein level was not changed.

CONCLUSIONp53(301-393)has a different localization in the p53 positive and p53 negative cells and could inhibit mitosis and cause the cell cycle arrest in G2/M.

Green Fluorescent Proteins ; genetics ; metabolism ; HeLa Cells ; Humans ; Immunoblotting ; Microscopy, Fluorescence ; Mitosis ; genetics ; physiology ; Mutant Proteins ; metabolism ; physiology ; Mutation ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; physiology

OBJECTIVETo investigate the role of Ser 219 phosphorylation of TRF1 (telomere repeat binding factor 1) in regulation of cell cycle.

METHODSThe mimicking phosphorylation mutant (TRF1S219D-GFP) and the non-phosphorylatable mutant (TRF1S219A-GFP) were constructed; the mutant genes and corresponding proteins were checked by sequencing and Western blot, respectively. Immunofluorescence staining was performed to detect the localization of mutants in HeLa cells. Cell cycle was analyzed by flow cytometry and ATM level was evaluated by immunoblotting.

RESULTSThe mutant genes were verified by direct sequencing and protein expression of GFP-tagged mutants was confirmed by immunoblotting.TRF1S219A-GFP and TRF1S219D-GFP were both localized in telomere of HeLa cells. Moreover, overexpression of TRF1-GFP or TRF1S219A-GFP resulted in an accumulation of HeLa cells in G2/M (P<0.05). The protein level of ATM was increased when overexpression the wide type or mutants.

CONCLUSIONThe Ser 219 phosphorylation of TRF1 by ATM could result in cell cycle arrest in G2/M, which is related to overexpression of TRF1.

Ataxia Telangiectasia Mutated Proteins ; Cell Cycle ; genetics ; physiology ; Cell Cycle Proteins ; metabolism ; DNA-Binding Proteins ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; HeLa Cells ; Humans ; Immunoblotting ; Microscopy, Fluorescence ; Mutation ; Phosphorylation ; Protein-Serine-Threonine Kinases ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Serine ; genetics ; metabolism ; Telomeric Repeat Binding Protein 1 ; genetics ; metabolism ; physiology ; Transfection ; Tumor Suppressor Proteins ; metabolism