Objective To identify the genotypes of ESBLs-producing Klebsiella pneumoniae isolates from the First Affiliated Hospital,Shantou University Medical College.Methods The MICs of 10 antibiotics were determined by agar-dilution against the clinical isolates of ESBLs-producing K.pneumoniae.PCR were performed with specific primers for blaTEM,blaSHV, blaCTX-M and blaOXA respectively.PCR products were cloned and sequenced.Results The results of PCR showed that a- mong the 83 strains of ESBLs-producing K.pneumoniae,75 were positive for blaTEM,41 positive for blaSHV,25 poitive for blaCTX-M,9 positive for hlaOXA.Three genotypes were found in 13 strains(15.7%),2 genotypes in 59 strains (71.1%) and single genotype in only 11 strains(13.2%).The genes of CTX-M-3,TEM-1 and SHV were found co-existent in 9 strains. The strains carrying 2 or 3 ESBL genes were more resistant to antibiotics than those carrying only 1 ESBL gene.Conclusions The genotypes of ESBLs-producing Klebsiella pneumoniae in this hospital are blaTEM,blaSHV,blaCTX-M and blaOXA. Most strains carry 2 or 3 ESBL genes.

Aged ; Female ; Hodgkin Disease ; diagnosis ; pathology ; Humans ; Reed-Sternberg Cells ; pathology

OBJECTIVETo improve inversion-polymerase chain reaction (I-PCR) in detection of factor VIII (FVIII) intron 22 inversion for gene diagnosis and prenatal diagnosis of hemophilia A (HA).

METHODSThe modified I-PCR was applied to detect FVIII intron 22 inversion in 8 families with HA. The prenatal diagnosis was performed for 2 pregnant women in the families. The fetal blood samples were obtained by cordocentesis at 22 and 26 weeks gestation respectively.

RESULTSFour of 8 HA families were diagnosed to be FVIII intron 22 inversion. Two fetuses were identified to be normal and one of them was born normal.

CONCLUSIONThe modified I-PCR enables the gene diagnosis and prenatal diagnosis of FVIII intron 22 inversion more accurately and rapidly.

Chromosome Inversion ; Factor VIII ; genetics ; Female ; Hemophilia A ; diagnosis ; genetics ; Humans ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis

AIM: To explore correlation,consistency and comfort between traditional tear film examination methods and Oculus Keratograph.METHODS: A retrospective study.Totally 101 cases (101 eyes) were diagnosed myopia and then accepted LASEK (laser epithelial keratomileusis).Non-invasive tear film break-up time(NIBUT),lower tear meniscus height(LTMH) were measured with Oculus Keratograph,fluorescein tear film break-up time(fl-BUT) and Schimer Ⅰ test (SⅠt) were performed on all cases.The correlations analysis between NIBUT and fl-BUT,LTMH and SⅠt were performed by Spearman rank correlation,consistency check between NIBUT and fl-BUT by Bland-Altman analysis.Visual analogue scale(VAS) was applied on evaluating the comfort of two kinds of examination methods.RESULTS: LTMH and SⅠt showed positive correlation (rs=0.346,P=0.001).NIBUT and fl-BUT showed positive correlation (rs=0.393,P=0.001),95% consistency limits range-9.62 to 14.18 in Bland-Altman Figure.There was significant difference between VAS of NIBUT and VAS of fl-BUT(z=-2.324,P=0.020).There was significant difference between VAS of LTMH and VAS of SⅠt (z=-8.845,P=0.001).CONCLUSION: Oculus Keratograph can objectively measure NIBUT and LTMH,and was more comfortable than traditional tear film examination methods.It can effectively assess tear film function before corneal refractive surgery.

Objective To investigate clinically and in laboratory a genealogical tree with hemoglobin H disease Combining Hemoglobin Q-Thailand and Hemoglobin E Disease.Methods Genealogical laboratory studies were carried out with the following methods:hemoglobin electrophoresis, various biochemical determinations,and DNA analysis.Results Father's genotype of ?-THAL:??/?~Q ?~(4.2); genotype of ?-THAL:?E/N;phenotype:minor ?-THAL carrier combining Hb Q and Hb E multiple heterozygote;mother' s genotype of ct-THAL:--~(SEA)/??;genotype of ?-THAL:?n/?n.According to comprehensive analysis,mother's phenotype:minor ?-THAL,complex minor ?-THAL carrier combining Hb F ? Initial sign of ?-THAL genotype:--~(SEA)/?~Q ?~(4.2);phenotype:deletion type Hb H genotype disease;?- THAL genotype:?E/?E;phenotype:? E homozygote.According to comprehensive analysis:deletion type Hb H combining HbE multiple heterozygote.Youger brother's ?-THAL genotype:--~(SEA)/?~Q ?~(4.2);?-THAL genotype:?n/?n;phenotype:deletion type Hb H disease.Both mother and her youngest son have G6PD deficiency.Conclusions Guangdong Province is an area with high morbidity of ?-THAL and ?-THAL,Hb E and Hb Q as well as G6PD deficiency.There may be some correlation between Hb E and Fib Q in term's of the high morbidity of regional Hb,but the two types of Hb combining Hb H disease are rare in China and the world in point of nonhomologous chromosome.Attention should be paid to the problems of double heterozygote of ?-THAL complex ?-THAL,and THAL complex G6PD deficiency.Data from the study have enriched the scientific information of molecular genetics of erythroeyte thalassemia and of molecular pathology with important significance in genetics guidance and clinical treatment for patients.

Adult ; Carcinoma ; virology ; Cervical Intraepithelial Neoplasia ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; In Situ Hybridization ; Middle Aged ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; virology

OBJECTIVETo obtain recombinant fusion protein HSA (human serum albumin)-PTH(1-34) in Pichia pastoris.

METHODSHSA and PTH(1-34) cDNA were obtained with PCR and the DNA segments were cloned into vector pPIC9 with linker. The linearized plasmids were transformed GS115 competent cells treated with LiCl, and mut+ transformants were screened on MD plate. With AOX promoter and alpha-MF signal sequences leading, fusion protein was expressed in GS115. PCR and SDS-PAGE were employed to confirm the integration and expression of HSA-PTH(1-34). The fusion protein was identified by Western blotting and classical adenylate cyclase assay.

RESULTThe PCR results showed that the gene of HSA-PTH(1-34) was integrated into GS115 genome. Western bolt approved the existence of two domains of HSA and PTH(1-34). The bioactivity assay in rabbit cortical membranes indicated that HSA-PTH (1-34) activated adenylate cyclase, but the activity was lower than that of the synthetic PTH(1-34).

CONCLUSIONActive fusion protein HSA-PTH (1-34) is successfully expressed in Pichia pastoris.

Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Peptide Fragments ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Serum Albumin ; biosynthesis ; genetics ; Teriparatide ; analogs & derivatives

OBJECTIVEThe aim of this study was to access the relationship of osteolytic bone metabolic markers such as serum type I collagen carboxy-terminal telopeptide (sICTP), N-terminal cross-linked telopeptides of type I collagen (uNTx), urinary pyridinoline (uPyd) with the therapeutic effect in breast cancer patients with bone metastases.

METHODS120 patients with breast cancer were included in this study. The levels of sICTP, uNTx and uPYD were measured by ELISA assay. The differences were compared between patients with and without bone metastasis. The patients with bone metastasis were treated and followed up as clinically indicated.

RESULTSThe levels of all above mentioned biomarkers in patients with bone metastasis were significantly higher than that in patients without bone metastasis (P < 0.01). A significant correlation was found between each two markers (r > 0.5, P < 0.01). The biomarkers were examined again in 45 patients with bone metastasis after treatment to evaluate the treatment response. The median follow-up was 10 months. Based on clinical evaluation criteria, 25 patients were responders and 20 were non-responders. For responders, after 3 months treatment, the levels of the three bone markers were significantly reduced (P = 0.025, P < 0.001, P < 0.001). But for non-responders, with progression of bone lesions, the levels of the three markers were significantly raised (P = 0.011, P = 0.002, P = 0.002). By means of multiple logistic regression with stepwise selection, the uPyd and uNTx activities were closely correlated with treatment response (OR = 17.0, P = 0.019; OR = 16.7, P = 0.015), however, the sICTP did not show any correlation with treatment response P = 0.841).

CONCLUSIONThe levels of sICTP, uNTx and uPyd may be used as indicators in assessment of the effect of antiresorptive treatment and evaluation of prognosis in breast cancer patient with bone metastases.

Adult ; Aged ; Amino Acids ; urine ; Biomarkers, Tumor ; metabolism ; Bone Neoplasms ; drug therapy ; metabolism ; secondary ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; Collagen ; urine ; Collagen Type I ; blood ; Disease Progression ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Peptides ; blood ; Remission Induction

Animals ; Fatty Liver ; drug therapy ; metabolism ; prevention & control ; Liver ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Taurine ; therapeutic use