The recombination clones contained CFP, LTB-ST foreign gene were screening by PCR using individual bacterial colonies as template, the aimed band was amplified from positive clones, the result was as well as plasmid PCR. The selecting of agrobacterium transformed with recombination plasmid could also use this method of PCR screening of individual bacterial colonies. The result of individual bacterial colonies PCR was as well as that of PCR using bacterial solution as template. It showed that the method individual bacterial colonies PCR was an efficient, easy one that characterized recombination clones.

Objective To research the method and effect about the bronchoscopic lung volume reduction by ?-cyanoac- rylate in the therapy of chronic obstructive pulmonary disease (COPD).Methods The 14 patients had been examined bosoms by CT before the operation and determined the type of emphysema and the distributing of pneumatocele,had blood gas analyzed and pulmonary function checked.The operation was carried through trachea cannula and intravenous anesthe- sia.When the bronchoscope came to the goal bronchia,we infused the meglumine diatrizoate through the biopsy orifice and approved the location of pneumatocele forward。Then,we infused erythromycin and ?-cyanoacrylate in turn through the biopsy orifice by silica del tube.Results The 3 pneumothorax patients had been removed the drainage tube in 3 days af- ter the operation.8 cases had been counterchecked sternite in one week and the pneumatocele was just like before,among which,1 case developed exudation.1 case had shown pleural thickening in the right-up lung counterchecked sternite 9 months later.1 case been checked the pulmonary function,the FEV_1 enhanced from 24.7% pred before operation to 32. 9% pred after operation one week.3 cases felt polypnea improved greatly and 7 cases felt polypnea improved a little.Con- clusion The bronchoscopic lung volume reduction by ?-cyanoacrylate is a safe,effective and economical method in the therapy of COPD.

Administration, Inhalation ; Adrenergic beta-Agonists ; adverse effects ; Child ; Extremities ; Humans ; Male ; Paralysis ; chemically induced ; Terbutaline ; administration & dosage ; adverse effects

Endoscopy ; Female ; Humans ; Middle Aged ; Nose ; surgery ; Oculomotor Muscles ; surgery

Adolescent ; Bile Reflux ; etiology ; Child ; Child, Preschool ; Female ; Gastritis ; etiology ; Gastroscopy ; Helicobacter Infections ; complications ; diagnosis ; Helicobacter pylori ; Humans ; Male

OBJECTIVETo study the association of FCGR2A gene single nucleotide polymorphism (SNP) rs1801274 with Kawasaki disease (KD) susceptibility and the efficacy of intravenous immunoglobulin (IVIG) therapy in Han Chinese children.

METHODSThirty-five KD children and 25 age-and gender-matched healthy children (control group) were enrolled in the study. Polymerase chain reaction (PCR) and gene sequence analysis were applied to detect SNP of FCGR2A gene rs1801274. These KD patients were classified into two subgroups based on the presence of coronary artery lesion (CAL) following IVIG therapy: CAL (n=13) and non-CAL (n=22).

RESULTSFCGR2A gene SNP rs1801274 was detected in all subjects, including three genotypes (AA, AG and GG). For FCGR2A gene SNP rs1801274, there were significant differences in the genotype and allele frequencies between the KD and control groups (P<0.05), and significant differences in the genotype and allele frequencies were also found between the CAL and non-CAL subgroups (P<0.05). A allele and AA genotype were linked to an increased risk of KD susceptibility (A allele: OR=3.39, 95%CI:1.53-7.50; AA genotype: (OR=4.93, 95%CI:1.61-15.1). Both AG (OR=5.43, 95%CI:1.06-27.8) and G allele (OR=4.88, 95%CI:1.44-16.5) were linked to an increased risk of CAL in KD children.

CONCLUSIONSPolymorphism of the FCGR2A gene SNP rs1801274 is one of the important factors probably influencing susceptibility to KD and efficacy of IVIG therapy on KD in Han Chinese children.

Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; genetics ; Polymorphism, Single Nucleotide ; Receptors, IgG ; genetics

OBJECTIVETo observe the effect of low molecular weight heparin calcium (LMWHC) combined Compound Danshen Injection (DI) on nephrotic syndrome patients with early onset severe preeclampsia.

METHODSTotally 80 nephrotic syndrome patients with early onset severe pre-eclampsia were randomly assigned to four groups voluntarily, i.e., Group A (22 cases, treated by magnesium sulfate), B (19 cases, treated by magnesium sulfate plus LMWHC), C (21 cases, magnesium sulfate plus DI), D (18 cases, magnesium sulfate plus LMWHC and DI). Umbilical arterial S/D ratios, amniotic fluid index (AFI), prolonged gestational age, placenta weight, neonatal weight, and Apgar score were compared among the four groups.

RESULTSCompared with before treatment in the same group, umbilical arterial S/D ratios decreased in the four groups (P <0. 05). AFI decreased in Group A, while it increased in Group B, C, and D (P<0. 05). Compared with Group A at the same time point, umbilical arterial S/D ratios decreased, and AFI increased in Group B, C, and D (P <0. 01 , P <0. 05). Prolonged gestational age and neonatal weight were increased in Group B, C, and D (P <0. 01, P <0. 05). Placenta weight were increased in Group B and D (P <0. 05). Apgar scores at 1 and 5 min were improved in Group D (P <0. 05). Compared with Group B and C at the same time point, umbilical arterial S/D ratios decreased, and AFI increased in Group D (P<0. 05). Compared with Group B, prolonged gestational age and placenta weight were decreased in Group C, but prolonged gestational age and placenta weight were increased in Group D (P <0.05). Compared with Group C, prolonged gestational age, placenta weight, and neonatal weight were increased in Group D (P <0. 05).

CONCLUSIONTreatment of nephrotic syndrome patients with early onset severe pre-eclampsia by LMWHC combined DI could prolong gestational ages, obviously improve prenatal outcomes, with better effect obtained than using any of them alone.

Calcium ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gestational Age ; Heparin, Low-Molecular-Weight ; therapeutic use ; Humans ; Magnesium Sulfate ; Nephrotic Syndrome ; drug therapy ; Phenanthrolines ; Pre-Eclampsia ; drug therapy ; Pregnancy ; Salvia miltiorrhiza

OBJECTIVETo investigate the precise locations of the blood vessels and nerves surrounding the seminal vesicles (SV) in men and provide some anatomical evidence for SV-related minimally invasive surgery.

METHODSWe observed the courses and distribution of the blood vessels and nerves surrounding SVs and obtained the data for positioning the SV neuroplexes in 20 male pelvises.

RESULTSOne branch of the neuroplexes was distributed to the SVs bilaterally with the neurovascular bundles, (2.85 ± 0.18) cm from the median sulcus of the prostate (MSP), while another branch ran through the Denonvillier fascia behind the SV, (0.81 ± 0.06) cm from the MSP. The arterial SVs (ASV) originated from the inferior vesical artery and fell into 4 types, 55% going directly to the SVs as one branch, 15% running between the SV and the ampulla of the deferent duct as another branch, 25% downward as 2 branches to the SV and between the SV and the ampulla of the deferent duct respectively, and 5% as the other ASVs. The shortest distance from the ASV through the prostatic neuroplexus to the posterior SV was (1.08 ± 0.09) cm.

CONCLUSIONIn SV resection, neuroplexus injury can be reduced with a bilateral distance of < 2.85 cm and a posterior distance of < 0.81 cm from the MSP, and so can bleeding by vascular ligation between the SV and the ampulla of the deferent duct.

Biopsy ; Humans ; Male ; Prostate ; blood supply ; innervation ; Seminal Vesicles ; blood supply ; innervation ; Vas Deferens ; blood supply ; innervation

Objective To explore the induced effects of colchicine in different concentration and times on Foeniculum vulgare polyploidy.Methods Seed germination rate and mutation rate,morphology of radicle induced mutants,chromosome number,tissue structure,and content and component of essential oil,which induced by colchicine in different concentiation and times were investigaled using the method of soaking seeds.Results The results showed that 0.13% of colchicine concentration and 24 h inducing time had the best induction on F.vulgare polyploidy.Compared to the control,the radicle of mutant was thicker,the chromosome number increased clearly,and the cell number in radicle manifolded distinctly.Among the four main components of essential oil,the contents of Dill apiol in anamorphosis radicle was a little lower than that of the control,but the contents of limonene,(E)-anethole,and camphene in anamorphosis radicle were all remarkably higher than that of the control after induced by colchicine.ConclusionThe content of main essential oil components in fennel mutant induced by colchicine is remarkably enhanced.The study provides the theoretical basis for the breeding of F.vulgare new varieties of high oil content.

This study aims to construct a eukaryotic expression system for envelope gene of Jaagsiekte sheep retrovirus, observes its localization in 293T cells, and investigates the potential in inducing malignant transformation of NIH3T3 cells. By RT-PCR, the full-length cDNA of envelope gene of Jaagsiekte sheep retrovirus (exJSRV-env) was amplified from the extract of naturally infected sheep lung. The clone of target gene was sub-cloned into eukaryotic expression system pEGFP-C1, and validated by PCR, restriction endonuclease, and sequencing. Bioinformatic analysis concerning biological function and cellular localiza tion of exJSRV-env was also performed. The recombinant clone of exJSRV-env was transfected into 293T cells and NIH3T3 cells by Lipofectamine LTX. The expression and celluar localization in 293T cells were validated by confocal microscopy. Soft agar colony formation assay was employed to test the anchorage-independent growth of NIH3T3. DNA sequencing and restriction enzyme digestion with Kpn I and Hind III indicated the correct construction of the recombinant plasmid, which was named pEGFP-C1/exJSRV-env. Amino acid sequence alignment of exJSRV-env with reference sequences found 85%-100% homogeneity. A YRNM motif was discovered at the cytoplasmic tail of envelope gene, which is exclusively found in exogenous viruses. Phylogenetic tree analysis showed that our clone of exJSRV-env clustered closely with pathogenic exogenous Jaagsiekte sheep retroviruses. Fluorescence microscopy indicated typical membrane localization of exJSRV-env protein. NIH3T3 cells transfected with exJSRV-env lost contact inhibition, and acquired colony forming ability in soft agar. This study indicated that envelope protein of Jaagsiekte sheep retrovirus can induce malignant transformation of mouse fibroblast cell NIH3T3. Discoveries of this study provide a basis for further structural and functional research on Jaagsiekte sheep retrovirus envelope protein.
Amino Acid Sequence ; Animals ; Betaretrovirus ; chemistry ; classification ; genetics ; physiology ; Cell Transformation, Viral ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; Molecular Sequence Data ; NIH 3T3 Cells ; Phylogeny ; Retroviridae Infections ; veterinary ; virology ; Sequence Alignment ; Sheep ; Sheep Diseases ; virology ; Transformation, Genetic ; Tumor Virus Infections ; veterinary ; virology ; Viral Envelope Proteins ; chemistry ; genetics ; metabolism