Objective To explore the relationship of the ratio of plasma adrenomedullin(AM)and endothelin-1(ET-1)with serum concentration of neuron-specific enolase(NSE)in full-term neonates with hypoxic-ischemic encephalopathy(HIE).Methods Plasma concentrations of AM,ET-1 and serum NSE from 32 full-term neonates with HIE were detected by radioimmunoassay(RIA)on the 1,3 and 7 d after parturition,30 neonates in the corresponding periods in our hospital were employed as controls.The infants with HIE were divided into mild,moderate or severe group in terms of diagnostic standard of HIE.Results 1.Plasma concentrations of AM and ET-1 in newborns with mild,moderate or severe HIE were significantly higher than that of control group at 1 d after life with a decline from 3-7 d(Pa

The aim of this study was to clone human RHD gene and to investigate its expression in transduced K562 cells. Total RNA was extracted from reticulocyte of cord blood. RHD and RHCE genes were amplified using RT-PCR method. The amplified products were cloned into pGEM-T plasmid by TA ligation and several clones were screened by direct sequencing method in order to obtain the RHD gene. RHD gene was subcloned into pcDNA3.1(-) expression vector, then the recombined plasmids were transduced into K562 cells with superfect transfection reagent kit. Finally transcription and expression of RHD gene in K562 cells were detected. The result showed that RHD gene has been cloned sucessfully, the inserted sequence and direction of RHD cDNA in its recombined pcDNA3.1(-) vector were identified using enzyme cutting and sequencing method. After transduced with recombined pcDNA3.1(-) vector, K562 cells could transcribe RHD mRNA in its cytoplasm and express RhD antigen on its membrane surface. In conclusion, RhD antigen can expressed in K562 cells with RHD cDNA transduction, and the expression system in vitro may be helpful to further investigate the molecular basis of RhD variants.
Base Sequence ; Cell Membrane ; metabolism ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Gene Expression ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics ; Rh-Hr Blood-Group System ; biosynthesis ; genetics ; Sequence Analysis, DNA ; Transfection

Long-term excessive iodine intake resulted in an increased TT_4 level and a decreased TT_3 level in maternal serum,meanwhile,hepatic and renal type 1 deiodinase activity decreased dose-dependently.A significant reduction in type 2 deiodinase ( D2 ) activity of 12.5 d placenta was found in 3.0 mg/L or above groups.For 19.5 d uterus,D2 activity decreased and type 3 deiodinase activity increased.The results suggest that excessive iodine has an effect on the embryonic development by regulating maternal-fetal thyroid hormone metabolism.

OBJECTIVETo summarize and analyze the clinical features of bladder tumor patients who received transurethral partial cystectomy by 2 microm continuous wave laser, in 1 year post operation follow-up visits.

METHODSFrom December 2007 to May 2008, 47 bladder carcinoma patients were treated with 2 microm laser transurethrally under sacral block. Operation characteristics, operation time, intraoperative hemorrhages and postoperative complications, and pathology staging of the tumor were observed and postoperative follow-up visits were performed.

RESULTSAll of the operation procedures were successful. The surgery time was 5 to 15 minutes. Blood loss in the operation was minimal. There was no obturator nerve reflection, and no hemorrhaging was detected after the operation. The pathological stages can be judged correctly with the obtained specimens. There was one case with peritoneum perforation. The patients received 12 to 17 months of postoperative follow-up visits, and there was no recurrence at the resection site. The survival rate was 100%.

CONCLUSIONSTransurethral partial cystectomy in the treatment of bladder tumor by 2 microm continuous wave laser is a safe, efficient and effective method. The tumor and all the basal part of bladder wall could be excised completely and the pathological stages can be judged correctly using these specimens to fulfill partial cystectomy for the treatment of bladder carcinoma.

Adult ; Aged ; Aged, 80 and over ; Cystectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Laser Therapy ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Urinary Bladder Neoplasms ; surgery

To investigate the molecular genetics basis for one para-Bombay phenotype, the red blood cell phenotype of the proband was characterized by standard serological techniques. Exon 6 and 7 of ABO gene, the entire coding region of FUT1 gene and FUT2 gene were amplified by polymerase chain reaction from genomic DNA of the proband respectively. The PCR products were purified by agarose gels and directly sequenced. The PCR-SSP and genescan were performed to confirm the mutations detected by sequencing. The results showed that the proband ABO genotype was A(102)A(102). Two heterozygous mutations of FUT1 gene, an A to G transition at position 682 and AG deletion at position 547-552 were detected in the proband. A682G could cause transition of Met-->Val at amino acid position 228, AG deletion at position 547-552 caused a reading frame shift and a premature stop codon. The FUT2 genotype was heterozygous for a functional allele Se(357) and a weakly functional allele Se(357), 385 (T/T homozygous at position 357 and A/T heterozygous at 385 position). It is concluded that the compound heterozygous mutation--a novel A682G missense mutation and a 547-552 del AG is the molecular mechanism of this para-Bombay phenotype.
ABO Blood-Group System ; genetics ; China ; DNA Mutational Analysis ; Female ; Fucosyltransferases ; genetics ; Genotype ; Humans ; Male ; Mutation ; Mutation, Missense ; Pedigree ; Phenotype ; Sequence Deletion

OBJECTIVETo explore the changes of surface antigen and function of rituximab on dendritic cells derived from patients with Primary immune thrombocytopenia (ITP) to further understand the effective mechanism of immunotherapy.

METHODSThe peripheral blood mononuclear cells (PBMCs) were isolated from remission patients with ITP before and after low-dose rituximab infusion, and the PMNCs were stimulated for 5 days by rhGM-CSF and rhlL-4 in 5% CO2 air at 37°C incubator. Then all of DCs were cultured with TNF-α for 48 hours. The morphology of DCs was monitored under inverted microscope daily, and the surface antigens of the DCs were analysed by flow cytometry, meanwhile the levels of IL-12p70 and TGF-β1 in supernatants were detected by ELISA, mix lymphocyte reaction was performed by MTT assay.

RESULTS(1) Rituximab-treated-DCs showed no obvious tree-like protruding compared with untreated-DCs. The former cells were small and most of nucleus were centric. (2) The expressions of HLA-DR, CD80, CD83 and CD86 on rituximab-treated-DCs \[56.37 ± 3.95)%, (36.41 ± 2.82)%, (30.45 ± 4.61)% and (41.98 ± 4.17)%, respectively\] were significantly lower than those untreated-DCs \[(73.71 ± 7.61)%, (55.14 ± 7.30)%, (80.91 ± 7.09)% and (59.03 ± 3.43)%, respectively\](all P < 0.05), the concentration of IL-12p70 was significantly lower, \[(66.87 ± 4.29)% vs (50.17 ± 14.52)%\], while that of TGF-β1 \[(9.70 ± 0.31)%\] higher than the untreated-DCs \[(2.70 ± 0.36)%\] (P < 0.05). (3) The abilities to activate T cells proliferation of rituximab-treated-DCs reduced compared with untreated-DCs.

CONCLUSIONThe surface antigen of ITP-DCs and the concentration of IL-12p70 reduced after the low-dose rituximab infusion. The abilities to activate T cells proliferation reduced while the concentration of TGF-β1 increased. Rituximab may achieve its therapeutic effect on ITP by downregulating the immunoreactivity of DCs.

Antibodies, Monoclonal, Murine-Derived ; administration & dosage ; therapeutic use ; Cell Proliferation ; Cells, Cultured ; Dendritic Cells ; cytology ; metabolism ; secretion ; Female ; Humans ; Interleukin-12 ; metabolism ; Lymphocyte Activation ; Male ; Rituximab ; T-Lymphocytes ; immunology ; Thrombocytopenia ; drug therapy ; immunology ; metabolism ; Transforming Growth Factor beta1 ; metabolism

RHD gene has different alternative transcripts. This study was aimed to construct expression vector of normal mRNA, DEL9 and DEL89 transcripts from RHD gene. Total RNA was extracted from Rh(D) positive umbilical blood cells of newborn. Intact RhD cDNA, DEL9 and DEL89 transcripts were obtained by one-step and two-step RT-PCR, respectively. The obtained products were cloned into pCR4 TOPO sequencing vector for choosing the right transcript. RHD gene was amplified again from the sequencing plasmid DNA, and then subcloned into pcDNA3.1/V5-His TOPO expression vector; DEL9 and DEL89 were cloned into the expression vector directly. Gene sequence and direction were identified by sequencing. The results showed that the sequence and direction of target genes were right, thus these 3 different expression vectors were correctly constructed. It is concluded that expression vector is constructed from different transcripts of RHD gene, which lays a foundation for further exploring the membranous protein expression of Rh(D) antigen.
Cloning, Molecular ; Gene Expression ; Genetic Vectors ; Humans ; Molecular Sequence Data ; RNA, Messenger ; genetics ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

OBJECTIVETo investigate the resistance genes and antibiotic resistance patterns against beta-lactams in Pseudomonas aeruginosa prevalent in burn ward.

METHODSK-B method was performed to test bacterial resistance patterns against 9 species of beta-lactams in Pseudomonas aeruginosa isolated from wounds and dressings of the patient in burn wards. Seven species of resistance genes against beta-lactams were detected with PCR. Tazobactam-inhibited piperacillin resistance test was performed to study whether the above strains produce extended spectrum beta-lactams.

RESULTSAll 12 strains of bacteria with resistance genes detected were resistant to penicillin and cephalosporins (100%), among them 11 were resistant to all antibiotics. Tazobactam-inhibited piperacillin resistance test demonstrated that all strains with resistance genes were ESBLs.

CONCLUSIONHigh incidence of beta-lactams resistance genes is found in Pseudomonas aeruginosa isolated from burn ward, and they have close relationship with the occurrence of multiple drug-resistance.

Burn Units ; Burns ; microbiology ; Genes, Bacterial ; Humans ; Pseudomonas aeruginosa ; drug effects ; genetics ; isolation & purification ; beta-Lactam Resistance ; genetics

OBJECTIVETo study risk factors for periventricular-intraventricular hemorrhage (PVH-IVH) in premature infants treated with mechanical ventilation.

METHODSA total of 205 premature infants who were admitted to the neonatal intensive care unit (NICU) and treated with mechanical ventilation between January 2009 and December 2011 were enrolled. They were classified into PVH-IVH and non-PVH-IVH groups according to the results of head ultrasonography performed at 3 to 7 days after birth. Single factor and multivariate logistic regression analysis were used to identify risk factors for PVH-IVH.

RESULTSSingle factor analysis indicated 9 factors associated with the development of PVH-IVH, including a gestational age of <32 weeks, a birth weight of <1500 g, intrauterine distress, severe asphyxia, vaginal delivery, maternal perinatal infection, premature rupture of membranes (PROM) at ≥8 hours, mechanical ventilation duration of ≥7 days and ventilator-associated pneumonia (VAP) (P<0.05). Multivariate logistic regression analysis showed that a birth weight of <1500 g (OR=2.665), intrauterine distress (OR=2.177), severe asphyxia (OR=5.653), maternal perinatal infection (OR=4.365) and VAP (OR=2.299) were independent risk factors for the development of PVH-IVH (P<0.05).

CONCLUSIONSVery low birth weight, intrauterine distress, severe asphyxia, maternal perinatal infection and VAP are closely associated with an increased risk of PVH-IVH in premature infants treated with mechanical ventilation. These clinical risk factors should be given more attention in the prevention of PVH-IVH.

Cerebral Hemorrhage ; etiology ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases ; etiology ; Intensive Care Units, Neonatal ; Logistic Models ; Male ; Pneumonia, Ventilator-Associated ; complications ; Prognosis ; Respiration, Artificial ; adverse effects ; Risk Factors