Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

A large number of bacteria were carried by mites parasitizing on animals and human,which including symbiotic and pathogenic bacteria.Mites were an important transmission media and could spread pathogenic bacteria.A total of 184 literatures were collected from database to analye diversity of bacteria carried by mites.There were about 105 species bacteria were carried by 94 mites.These bacteria belong to 9 phylums,22 orders,40 families and 55 genuses(including 17 pathogen and 20 opportunistic pathogen).In this paper,we reviewed the diversity of mites-associated bacteria,which could offer some data for investigation on the relationship between mites and mites-associate bacteria.

OBJECTIVETo achieve pregnancy with unaffected embryo using in vitro fertilization and embryo transfer (IVF-ET) and preimplantation genetic diagnosis(PGD) for the couples at risk of having children with beta-thalassemia.

METHODSA couple carrying different thalassemia mutations of codon 41/42 and codon IVS2 position 654 received standard IVF treatment and intracytoplasmic sperm injection, embryo biopsy, single cell polymerase chain reaction and DNA analyses, and only the unaffected or carrier embryos were transferred to uterus. Pregnancy confirmation, and prenatal diagnosis were done at 20 week's gestation.

RESULTSA total of 13 embryos were analyzed in the IVF cycle. PGD indicated that 2 were normal 18.1 , 3 were affected 27.3 , and 6 were carriers 54.5 ; diagnosis was not possible in 2. Three embryos were transferred to uterus on the third day after oocyte retrieval. Ultrasonography showed twin pregnancy with one blighted ovum. The prenatal diagnoses revealed that both fetuses were unaffected, one normal baby and one carrier were born.

CONCLUSIONThese studies represent the successful application of PGD for beta-thalassemia in China.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Male ; Mutation ; Pregnancy ; Pregnancy Outcome ; Preimplantation Diagnosis ; methods ; Prenatal Diagnosis ; methods ; beta-Thalassemia ; diagnosis ; genetics ; prevention & control

Adolescent ; Adult ; Aged ; Body Temperature ; Craniocerebral Trauma ; physiopathology ; therapy ; Female ; Humans ; Hypothermia, Induced ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the variance and drug resistance of pathogenic bacteria isolated from children with infectious diseases seen between 2001 and 2006 in Chengdu area.

METHODSA total of 2888 pathogenic bacterial strains isolated from children in Chengdu Children's Hospital from 2001 to 2006 were analyzed. Tests were performed according to the guidelines of National Committee for Clinical Laboratory Standards (NCCLS) of the United States.

RESULT(1) Of the 2888 strains, 1845 (63.9%) were Gram negative bacteria. The main pathogenic bacteria included Escherichia coli (Ec, 718 strain, 24.9%), Hemophilus (H, 476 strain, 16.5%), Streptococcus pneumoniae (Sp, 412 strain, 14.3%), Klebsiella pneumoniae (Kp, 369 strain, 12.8%), Staphylococcus aureus (Sa, 353 strain, 12.2%), Staphylococcus epidermidis (Se, 278 strain, 9.6%), Pseudomonas aeruginosa (Pa, 146 strain, 5.1%) and other non-zymocyte (Onz, 136 strain, 4.7%). (2) The common pathogens found in blood specimen were 158 strain, which included Se (78 strain, 49.4%), Ec (23 strain, 14.6%), Kp (17 strain, 10.8%), Sa (14 strain, 8.9%), Onz (14 strain, 8.9%), Sp (7 strain, 4.4%) and Pa (5 strain, 3.2%). (3) The number of common pathogens isolated from patients with lower respiratory tract infection was 2018, including Ec (441 strains, 21.9%), H (430 strains, 21.3%), Sp (368 strains, 18.2%), Kp (253 strains, 12.5%), Sa (207 strains, 10.3%), Se (149 strains, 7.3%), Pa (97 strains, 4.8%) and Onz (73 strains, 3.6%). (4) There were 120 strains of common pathogens isolated from urine specimens, including Ec (78 strains, 65%), Kp (25 strains, 20.8%), Onz (7 strains, 5.8%), Pa (5 strains, 4.2%) and Se (5 strains, 4.2%). (5) There were 497 strains of common pathogens found in pus specimens, including Ec 167 strains, (33.6%), Sa (126 strains, 25.4%), Se (46 strains, 9.3%), H (44 strains, 8.9%), Onz (37 stains, 7.4%), Kp (31 strains, 6.2%), Sp (26 strains, 5.2%) and Pa (20 strain, 4.0%). The trend of drug resistance of pathogenic bacteria to antibiotics deteriorated. The proportion of methicillin-resistant Staphylococcus aureus (MRSA) was 6.7% and the methicillin-resistant coagulase-negative Staphylococci (MRCNS) rate was 20% in 2001 - 2003. The total proportion of extended-spectrum beta-lactamase stains (ESBL(S)) in Ec and Kp was 21.8%, and the rate of beta-lactamase production stains of Hemophilus influenzae (Hi) was 19.4% in 2001 - 2003.The proportion of MRSA was 17.2% and the MRCNS rate was 70.2%, the total proportion of ESBL(S) in Ec and Kp was 43.8%, and the rate of beta-lactamase producing stains of Hi was 39.7% in 2004 - 2006.

CONCLUSIONThe distribution of common pathogenic bacteria seen in Chengdu Children's Hospital has changed and the trend of drug resistance of pathogenic bacteria to antibiotics deteriorated in recent three years. Regionally monitoring the changes in pathogenic bacteria and the trend of drug resistance to antibiotics is paramount in guiding the pediatric clinical treatment.

Anti-Bacterial Agents ; pharmacology ; Bacterial Infections ; epidemiology ; microbiology ; Child ; China ; epidemiology ; Drug Resistance, Multiple, Bacterial ; drug effects ; Humans ; Microbial Sensitivity Tests

OBJECTIVETo study the clinical significance of tidal breathing lung function test in 1-4 years old children with wheezing diseases.

METHODSA total of 141 1-4 years old children with wheezing diseases were enrolled as the observed groups (41 cases of asthma, 54 cases of asthmatic bronchitis, and 46 cases of bronchopneumonia). Thirty children without respiratory diseases were enrolled as the control group. All the recruits underwent tidal breathing lung function test. The observed groups underwent bronchial dilation test, and tidal breathing flow volume (TBFV) parameters were evaluated before and after bronchial dilation test.

RESULTSThe observed groups showed obstructive ventilatory disorder (65%) according to the TBFV loop, and their ratio of time to peak tidal expiratory flow (TPTEF) to total expiratory time (TE) and ratio of volume to peak expiratory flow (VPEF) to total expiratory volume (VE) were significantly lower than in the control group (P<0.05). The asthma subgroup had significantly improved TPTEF/TE and VPEF/VE after bronchial dilation test (P<0.05). Taking an improvement rate of ≥ 15% either for TPTEF/TE or for VPEF/VE as an indicator of positive bronchial dilation test, the bronchial dilation test had a sensitivity of 47% and a specificity of 84% in diagnosing asthma in 1-4 years old children. The positive rate was 28% among the children in the asthma subgroup with an TPTEF/TE ratio of ≥ 23% before bronchial dilation test, versus 65% in those with an TPTEF/TE ratio of <23%.

CONCLUSIONSObstructive ventilatory disorder is the main impairment of tidal breathing lung function in 1-4 years old children with wheezing diseases. Tidal breathing bronchial dilation test can reflect a reversal of airway obstruction to a certain extent. The sensitivity of bronchial dilation test for the diagnosis of asthma is not satisfactory in 1-4 years old children with wheezing diseases, but this test has a relatively high diagnostic value in children with severe airway obstruction.

Asthma ; diagnosis ; physiopathology ; Bronchitis ; diagnosis ; physiopathology ; Bronchopneumonia ; diagnosis ; physiopathology ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Respiration ; Respiratory Function Tests ; methods ; Respiratory Sounds ; diagnosis ; drug effects ; physiopathology