AIMTo determine whether adenoviral gene transfer of insulin like growth factor-1 (IGF-1) to the penis of streptozotocin (STZ)-induced diabetic rats could improve erectile capacity.

METHODSTHE STZ diabetic rats were transfected with AdCMV-betagal or AdCMV-IGF-1. These rats underwent cavernous nerve stimulation to assess erectile function and their responses were compared with those of age-matched control rats 1 to 2 days after transfection. In control and transfected STZ diabetic rats, IGF-1 expression were examined by reverse transcription polymerase chain reaction (RT-PCR), Western blot and histology. The penis beta-galactosidase activity and localization of the STZ diabetic rats were also determined.

RESULTSOne to two days after transfection, the beta-galactosidase was found in the smooth muscle cells of the diabetic rat penis transfected with AdCMV-betagal. One to 2 days after administration of AdCMV-IGF-1, the cavernosal pressure, as determined by the ratio of maximal intracavernous pressure-to-mean arterial pressure (ICP/MAP) and total intracavernous pressure (ICP), was increased in response to cavernous nerve stimulation. Transgene expression was confirmed by RT-PCR, Western blot and histology.

CONCLUSIONGene transfer of IGF-1 significantly increased erectile function in the STZ diabetic rats. These results suggest that in vivo gene transfer of IGF-1 might be a new therapeutic intervention for the treatment of erectile dysfunction (ED) in the STZ diabetic rats.

Animals ; Blood Glucose ; metabolism ; Body Weight ; Diabetes Mellitus, Experimental ; physiopathology ; Erectile Dysfunction ; prevention & control ; Genetic Therapy ; Insulin-Like Growth Factor I ; genetics ; Male ; Penile Erection ; physiology ; Penis ; enzymology ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Reference Values ; beta-Galactosidase ; metabolism

With rapid and efficient drug release, few side effects and excellent biodegradable properties, the reduction-sensitive carriers is not only the new hot point in the field of pharmaceutical research, but also the most promising intelligent drug carrier on clinical application. This paper reviews the latest research of reduction-sensitive drug and gene carriers, including the mechanisms of drug release and the synthesis of the reduction-sensitive conjugates, reduction-sensitive nano polymer micelles, nano vesicles, nano hollow microspheres, nano liposomes, as well as the characteristics and advantages of various kinds of carrier system. It will provide a theoretical basis for its further application.
Drug Carriers ; administration & dosage ; chemistry ; Micelles ; Microspheres ; Nanoparticles ; Oxidation-Reduction ; Pharmaceutical Preparations ; administration & dosage ; chemistry ; Polymers ; chemistry ; Sensitivity and Specificity

OBJECTIVESTo analyze the pathogenic bacterial's distribution and the drug resistance in the upper urinary tract stones, and to provide the information for choosing suitable antibiotics.

METHODSStone samples were taken for culture and for drug sensitivity test in 146 patients undergoing percutaneous nephrolithotomy between April 2007 and October 2008, and the results were analyzed.

RESULTSPathogens presented in 72 (49.3%) patients. There were 70 (86.4%) Gram-negative bacteria strains. Escherichia coli, Pseudomonas aeruginosa and Enterobacter cloacae were the predominant bacteria, accounted for 30.9%(25 strains), 23.5% (19 strains) and 12.3% (10 strains), respectively. There were 10 (12.3%) Gram-positive bacteria strains, the predominant bacteria was Staphylococcus epidermidis (6 strains), accounting for 7.4%. And there was 1 fungi strain (1.2%). Resistance to ampicillin/sulbactam (88.7%), ceftriaxone (81.3%) and ciprofloxacin (67.5%) was most commonly found in pathogen, and the rate of resistance to amikacin, imipenem and piperacillin/tazobactam were 8.6%, 10.0%, 10.0%, respectively. Erythromycylamine, teicoplanin, SMZ-TMP, nitrofurantoin were sensitive to Gram-positive bacteria.

CONCLUSIONSBacterial's distribution of upper urinary tract stones are multiple, and the majority pathogen is Gram-negative bacteria. A big variant resistance is found among different bacterium. The suitable antibiotics should be chosen according to the different bacterium in the patients who underwent percutaneous nephrolithotomy.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bacteria ; drug effects ; isolation & purification ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Kidney Calculi ; microbiology ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Retrospective Studies ; Ureteral Calculi ; microbiology ; Young Adult

OBJECTIVEReal-time RT-PCR, cell culture and embryonated eggs culture for influenza detection were compared by analyzing the data of influenza surveillance in Shenzhen in second half of 2009.

METHODS1092 clinical samples (throat swabs) collected during second half of 2009 were tested by real-time RT-PCR, cell culture and embryonated eggs culture, and the results were analyzed by statistical methods.

RESULTSThe positive rate were 54.21%, 27.11% and 16.21% using real-time RT-PCR, cell culture and embryonated eggs culture, and the sensitive were 100%, 50% and 29.9%. The lowest dilutions of virus detected by real-time RT-PCR were 10(-2) TCID50/ml.

CONCLUSIONThe sensitive of real-time RT-PCR was higher than culture and the specificity was also very high. It was more suitable for emergency detect. The sensitive of cell culture for H3N2 subtype was higher, and sensitive of embryonated eggs culture for type B was higher.

Animals ; Chick Embryo ; Humans ; Orthomyxoviridae ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Virus Cultivation ; methods

Objective To estimate the real number of novel influenza A(H1N1 ) infection in Beijing, 2009. Methods A multiplier model (Impact 2009 v 1.0 software) based on Monte Carlo approach was used to estimate the real number of novel influenza A (H1N1 ) based on the number of influenza-like illness (ILI) cases, novel influenza A(H1N1 ) positive rate among ILI cases and rate on clinical visit of ILIs in secondary and tertiary hospitals. Results There were 1.80 million (90%CI: 1.46-2.30) estimated novel influenza A (H1N1) cases in 2009 in Beijing with the rate of infection as 11.0%. One reported case would represent 167 real infections. The highest age groups of infection were 0-4 years and 5-14 years, being 32.5% and 33.3%, respectively. Conclusion Laboratory-confirmed infections with novel influenza A (H1N1 ) only represented a fraction of the total cases in a population, suggesting that it was imperative to estimate the real number of novel influenza A (H1N1) infection.

OBJECTIVETo observe the prevalence of hypertension and associated risk factors in the Guangxi Bai Ku Yao populations.

METHODSA total of 1170 subjects of Bai Ku Yao aged 15 and over were surveyed by a stratified randomized cluster sampling. Blood pressure, body height, weight, waist circumference, serum lipid and apolipoprotein levels were measured, and body mass index (BMI) were calculated, matched 1173 subjects of Han Chinese from the same region served as control.

RESULTSThe standardized prevalence of hypertension in Bai Ku Yao was significantly lower than that in Han (11.53% vs.16.79%, P < 0.01). The mean levels of systolic, diastolic blood pressure, and pulse pressure in Bai Ku Yao were also significantly lower than those in Han [(115.7 +/- 16.3) vs. (120.0 +/- 16.3) mm Hg (1 mm Hg = 0.133 kPa), P < 0.01; (74.1 +/- 9.4) vs. (75.9 +/- 10.4) mm Hg, P < 0.01; and (41.6 +/- 12.0) vs. (44.2 +/- 11.2) mm Hg, P < 0.01; respectively]. Hypertension was positively correlated with male, age, physical activity, BMI, waist circumference, and the intakes of total energy, total fat, and sodium, and negatively associated with education level in both ethnic groups (P < 0.05 - 0.01), but was positively associated with alcohol consumption only in Han. The rates of awareness, treatment and control of hypertension were significantly lower in Bai Ku Yao than those in Han population [(11.81% vs. 21.76%), P < 0.05; (5.51% vs. 12.95%), P < 0.05; and (2.36% vs. 8.29%), P < 0.05; respectively].

CONCLUSIONThe prevalence of hypertension was significantly lower in Bai Ku Yao population than in Han population and diet, low sodium intake, life style, and genetic factors might be responsible for the lower hypertension prevalence in Bai Ku Yao population.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Feeding Behavior ; Female ; Humans ; Hypertension ; epidemiology ; ethnology ; Life Style ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Young Adult

BACKGROUNDMonocytes and macrophages in atherosclerotic plaque lead to plaque instability. The aim of the study was to determine if plaque neovascularization led to inflammation.

METHODSPatients were consecutively enrolled if their carotid intimal media thickness was > 2 mm, as revealed by duplex ultrasound. The patients then underwent dynamic contrast enhanced magnetic resonance imaging (DCE MRI) and fluorine-18 fluorodeoxyglucose ((18)F-FDG) positron emission tomography combined with computed tomography (PET CT). A target to background ratio (TBR) of ≥ 1.25 or < 1.25 served as the cutoff point for the presence and absence of inflammation, respectively.

RESULTSTwenty-six patients underwent bilateral carotid DCE MRI and 24 patients also underwent PET CT. One hundred and fifty-five plaques were evaluated by both DCE MRI and PET CT. There was no significant difference in plaque morphology between the TBR ≥ 1.25 (n = 61) and TBR < 1.25 (n = 94) groups. No significant differences were found in plasma volume and transfer constant between the TBR ≥ 1.25 and TBR < 1.25 groups.

CONCLUSIONOur study did not find a significant correlation between plaque neovascularization and the aggregation of inflammatory cells.

Aged ; Aged, 80 and over ; Carotid Artery Diseases ; pathology ; Cell Aggregation ; Female ; Fluorodeoxyglucose F18 ; Humans ; Inflammation ; pathology ; Macrophages ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neovascularization, Pathologic ; Plaque, Atherosclerotic ; pathology ; Positron-Emission Tomography ; Tomography, X-Ray Computed

OBJECTIVETo filtrate breast cancer resistance protein (BCRP)-mediated resistant agents and to investigate clinical relationship between BCRP expression and drug resistance.

METHODSMTT assay was performed to filtrate BCRP-mediated resistant agents with BCRP expression cell model and to detect chemosensitivity of breast cancer tissue specimens to these agents. A high performance liquid chromatography (HPLC) assay was established, and was used to measure the relative dose of intracellular retention resistant agents. RT-PCR and immunohistochemistry (IHC) were employed to investigate the BCRP expression in breast cancer tissue specimens.

RESULTSMTT assay showed that the expression of BCRP increased with the increasing resistance of 5-fluorouracil (5-Fu) (P<0.05, n=3) in the cell model, while HPLC assay indicated that the intracellular retention dose of 5-Fu was significantly correlated with the expression of BCRP (r=-0.897, P<0.05, n=3). A total of 140 breast cancer tissue specimens were collected. BCRP-positive expression was detected in forty-seven specimens by both RT-PCR and IHC. As shown by MTT assay subsequently, the resistance index (RI) of 47 BCRP-positive breast cancer tissue specimens to 5-Fu was 7-12 times as high as that of adjacent normal tissue samples. BCRP expression was related to 5-Fu resistance (R2=0.8124, P<0.01).

CONCLUSIONResistance to 5-Fu can be mediated by BCRP. Clinical chemotherapy for breast cancer patients can be optimized based on BCRP-positive expression.

ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; metabolism ; Adult ; Antimetabolites, Antineoplastic ; pharmacology ; Chromatography, High Pressure Liquid ; Drug Resistance, Neoplasm ; Female ; Fluorouracil ; pharmacology ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasm Proteins ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo develop a specific SARS virus-targeted antibody preparation for emergent prophylaxis and treatment of SARS virus infection.

METHODSBy using phage display technology, we constructed a naive antibody library from convalescent SARS patient lymphocytes. To obtain the neutralizing antibody to SARS virus surface proteins, the library panning procedure was performed on purified SARS virions and the specific Fab antibody clones were enriched by four rounds of repeated panning procedure and screened by highthroughput selection. The selected Fab antibodies expressed in the periplasma of E. coli were soluble and further purified and tested for their binding properties and antiviral function to SARS virus. The functional Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell systems and their neutralizing activities were further determined.

RESULTSAfter four rounds of the panning, a number of SARS-CoV virus-targeted human recombinant Fab antibodies were isolated from the SARS patient antibody library. Most of these were identified to recognize both natural and recombinant SARS spike (S) proteins, two Fab antibodies were specific for the virus membrane (M) protein, only one bound to SARS-CoV nucleocapsid protein. The SARS-CoV S and M protein-targeted Fab or IgG antibodies showed significant neutralizing activities in cytopathic effect (CPE) inhibition neutralization test, these antibodies were able to completely neutralize the SARS virus and protect the Vero cells from CPE after virus infection. However, the N protein-targeted Fab or IgG antibodies failed to neutralize the virus. In addition, the SARS N protein-targeted human Fab antibody reacted with the denatured N proteins, whereas none of the S and M protein specific neutralizing antibodies did. These results suggested that the S and M protein-specific neutralizing antibodies could recognize conformational epitopes which might be involved in the binding of virions to cellular receptors and the fusion activity of the virus.

CONCLUSIONThe SARS-CoV spike protein and membrane proteins are able to elicite efficient neutralizing antibodies in SARS patients. The neutralizing antibodies we generated in this study may be more promising candidates for prophylaxis and treatment of SARS infection.

Amino Acid Sequence ; Animals ; Antibodies, Viral ; immunology ; Cercopithecus aethiops ; Humans ; Membrane Glycoproteins ; immunology ; Neutralization Tests ; Peptide Library ; Protein Binding ; Protein Engineering ; Recombinant Proteins ; immunology ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; immunology ; virology ; Spike Glycoprotein, Coronavirus ; Vero Cells ; Viral Envelope Proteins ; immunology ; Viral Matrix Proteins ; immunology

Objective To investigate the source of the first human case of avian influenza A (H5N1) infection in Beijing. Methods Interviewing the relatives of the case and other key persons, collecting and detecting samples of related biological, epidemiological and environmental data of the case were conducted. Later, the infection source was thoroughly investigated. Results The case ever contacted a slaughtered duck 5 days prior to the onset of illness, and the duck was bought from a stall of a wet market in Yanjiao area of Hebei province. Ten environmental samples were collected in this stall and the neighboring stall of the market. Another 6 samples were tested positive for H5N1 virus by PCR method, with 5 virus strains isolated. The whole-genome sequencing indicated that the amino acid homology between the H5N1 virus strains from the environment and the virus isolated from the case reached 99.8%-100%. Conclusion From both epidemiological and virological evidence, it was proved that the first human case of avian influenza A (H5N1) infection in Beijing was infected by a duck that carrying H5N1 virus the case contacted 5 days proceeding the onset of illness.