The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.
Animals ; Bungarus ; classification ; genetics ; Cytochromes b ; genetics ; DNA Primers ; genetics ; Drug Contamination ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Alignment ; Sequence Analysis, DNA

OBJECTIVEThis study was designed to investigate the effects of different oxygen inhalation modes on retinal vessels development in neonatal mice in order to provide experimental data for proper oxygen therapy for premature infants.

METHODSA total of 144 postnatal day (P) 7 C57BL/6J mice were randomly assigned into 6 groups according to different oxygen inhalation modes (n=24). Experimental group 1 was exposed to 30%, 40%, 50%, 60% and 75% oxygen in turn for one day respectively, followed by room air exposure for 5 days. Experimental group 2 was exposed to 75%, 60%, 50%, 40% and 30% oxygen in turn for one day respectively, followed by room air exposure for 5 days. Experimental group 3 was exposed to 75% oxygen for 5 days, followed by room air exposure for 5 days. Experimental group 4 was exposed to 75% oxygen for 5 days, 50% oxygen for 2 days and 30% oxygen for 2 days, then room air exposure for 6 days. The supplemental 75% oxygen and room air recovering was performed alternately for the mice in Experimental group 5 for 3 times and then room air exposure for 5 days. The Control group was exposed to room air for consecutive 10 days. The retinal vascular development and proliferation were evaluated by the retinal flat-mounts (ADPase stained retina) and cross-section.

RESULTSThe peripheral vascular pattern was clear, and a few avascular areas were seen in the Control group at P12. At P14 the avascular area disappeared. At P17, the entire vascular pattern became completely normal. In the Experimental groups 1, 3 and 5, the central vessels became tortuous and constricted and the central avascular area increased at P12. At P14, neovascularization was seen peaking at P17 in the Experimental groups 1, 3 and 5. In the Experimental group 4, the central avascular area increased and neovascularization was seen at P14, but the central avascular area was reduced and abnormal neovascularization disappeared, with slight constriction of the deep vessels, at P17. Five days later the vascular pattern became almost normal in the Experimental group 4. The retinal vascular form of the Experimental group 2 was similar to that of the Control group. The average number of neovascular nuclei extending into the vitreous per cross-section in the Experimental groups 1, 2, 3, 4, and 5 and the Control group was 49.50 +/- 1.36, 5.17 +/- 0.67, 47.68 +/- 4.70, 5.74 +/- 2.37, 29.15 +/- 2.48, and 1.22 +/- 0.20 respectively. There were significant differences between the Experimental groups 1, 3, 5 and the Control group (P < 0.05).

CONCLUSIONSThe effects of different oxygen inhalation modes on the retinal vessels development in neonatal mice were different. The obvious fluctuation of inhaled oxygen concentration and abrupt stop of supplemental oxygen after high levels of supplemental oxygen may severely affect the development of retina vascular, leading to the pathologic changes similar to retinopathy of prematurity.

Animals ; Female ; Humans ; Infant, Newborn ; Male ; Mice ; Mice, Inbred C57BL ; Oxygen Inhalation Therapy ; methods ; Retina ; growth & development ; Retinal Neovascularization ; Retinopathy of Prematurity ; etiology

BACKGROUNDRecent studies have shown that T helper type-2 (Th2) cells can induce the apoptosis of CD4+CD25+ Treg cells or resist the immunosuppressive effect of Treg cells. We hypothesize that an imbalance of Th2/Treg is present in patients with allergic asthma.

METHODSTwenty-two patients with mild asthma, 17 patients with moderate to severe asthma, and 20 healthy donors were enrolled. All patients were allergic to house dust mites. The proportion of peripheral blood CD4+CD25+ Treg cells and Th2 cells were determined by flow cytometry. The concentration of interleukin (IL)-10, transforming growth factor (TGF)-β and IL-4 in plasma was determined by enzyme linked immunosorbent assay. In these subjects, peripheral blood mononuclear cells from 17 mild asthmatic patients, 13 moderate to severe asthmatic patients and 14 healthy donors were acquired and expression of forkhead box P3 (Foxp3) and GATA-3 mRNA was detected by reverse-transcriptase polymerase chain reaction.

RESULTSCompared with healthy donors and patients with mild asthma, the percent of CD4+CD25+ Treg cells and plasma IL-10 levels were decreased in patients with moderate to severe asthma. There were no significant differences in Foxp3 mRNA expression among three groups, but a downward trend seen among patients with asthma. However, the percent of Th2 cells, IL-4 levels and expression of GATA-3 mRNA was markedly higher in patients with mild and moderate to severe asthma than in the control group. The ratio of Th2/Treg and their cytokines was increased in allergic asthma, especially for moderate to severe asthma. The ratio of GATA-3/Foxp3 mRNA was also increased in allergic asthma. In patients with moderate to severe asthma, the percentage of peripheral blood Treg cells was negatively correlated to the percentage of Th2 cells and IL-4 levels.

CONCLUSIONSThe decline of CD4+CD25+ Treg cells in patients with moderate to severe asthma may play an important role in progress of the disease. Furthermore, the deficiency of CD4+CD25+ Treg cells was associated with the over-expression of Th2 response.

Asthma ; etiology ; immunology ; Cytokines ; blood ; Forkhead Transcription Factors ; genetics ; GATA3 Transcription Factor ; genetics ; Humans ; RNA, Messenger ; analysis ; T-Lymphocytes, Regulatory ; immunology ; Th2 Cells ; immunology

OBJECTIVEThis study examined the status of health-related behaviors among rural residents and the factors influencing the practice of such behaviors.

METHODSOne thousand and ninety subjects aged 15 years or over in a rural community, Anhui Province, China were surveyed. A questionnaire was used to collect information on the health knowledge, attitude and behavior of the subjects. Information on health behavior included smoking, drinking, dietary habits, regular exercises, sleeping pattern and oral health behavior.

RESULTSThe prevalence of smoking and drinking in the male subjects was 46.5% and 46.9%, respectively. There was a positive significant association between smoking and drinking. Only 8.3% of all subjects ate three regular meals a day regularly. Among subjects who ate two meals a day, 89.7% did not have breakfast. Only 1.7% of subjects took part in regular exercise. About 85% of subjects slept 6 to 8 h per day. Only 38.4% of the respondents had the habit of hand washing before eating and after using the lavatory. 79.3% of the subjects brushed their teeth every day, and among them, only 10.6 percent brushed their teeth twice a day. Further analyses showed that 64.8% of subjects had 3-5 items of positive health behaviors out of 8 items and only 16.9% had six or more items. Logistical regression analyses suggested that better health behavior was affected by sex, age, years of education, income and health knowledge.

CONCLUSIONThe status of health behaviors among rural residents was generally poor. It is thus urgent to reinforce health education in rural communities in China.

Adolescent ; Adult ; Aged ; Alcohol Drinking ; epidemiology ; Attitude to Health ; China ; epidemiology ; Cross-Sectional Studies ; Diet ; Exercise ; Female ; Health Behavior ; Health Surveys ; Humans ; Male ; Middle Aged ; Oral Health ; Risk-Taking ; Rural Population ; Sleep ; Smoking ; epidemiology

0507JS60 virus was isolated from a pool of Culex sp. collected in Kashi, Xinjiang, which could be propagated stably on C6/36 cells and caused cytopathic effects continuously. Viral particles had no envelope and appeared round with diameter of about 55nm (n = 10). Capsomeres on the surface of capsid were clearly visible. Electrophoresis of viral genome showed a profile of 12 double stranded RNA (dsRNA) segments. Sequencing of the twelfth segment revealed the length of 760bp (GenBank ID: FJ157354). A single open reading frame (ORF) was found and encoded a protein of 174 amino acids with a molecular mass of 18.9kD. The nucleotide sequence had similarity over 89% with that of LNV, but the deduced amino acid sequence had similarity over 91% with that of LNV. A phylogenetic tree was constructed to compare the corresponding genetic sequences in Seadornavirus. The tree demonstrated that 0507JS60 virus lied in the same branch with LNV and more closely related to LNV-NE9712. 0507JS60 virus was identified as LNV, which was firstly isolated outside the Northeast of China.
Animals ; Cell Line ; China ; Culex ; virology ; Molecular Sequence Data ; Phylogeny ; Reoviridae ; classification ; genetics ; isolation & purification ; ultrastructure

0507BS3 virus was isolated from a mixed pool of Culex sp. and Anopheles sp. collected in Kashi, Xinjiang, China. 0507BS3 virus could cause cytopathic effects on C6/36 cells but not on Vero and BHK-21 cells. Viral particles had no envelope and appeared round with diameter of about 60 nm (n = 20). Viral capsid was composed of a single layer and a central core. Capsomeres on the surface of capsid were clearly visible. Electrophoresis of viral genome showed a profile of 10 double stranded RNA (dsRNA) segments. Sequencing of the tenth segment revealed the length of 964bp (GenBank ID: FJ150869). A single open reading frame (ORF) was found and encoded a protein of 275 amino acids with a molecular mass of 30.8kDa. The nucleotide sequence had no similarity with any other viral genomic sequences, but the deduced amino acid sequence significantly matched the polyhedrin genes of cytoplasmic polyhedrosis virus (CPV) in some sections. A phylogenetic tree was constructed to compare the polyhedrin gene sequences of all CPV types in GenBank. The tree demonstrated that 0507BS3 virus was only distantly related to the other CPV types and belonged to a new CPV type.
Animals ; Cell Line ; China ; Culicidae ; virology ; Phylogeny ; Reoviridae ; classification ; genetics ; isolation & purification ; ultrastructure

BACKGROUNDIncreasingly, evidence from population, clinic-based and laboratory studies supports an independent association between obstructive sleep apnea syndrome (OSAS) and an increased risk of type 2 diabetes; however, this observation has yet to be replicated in China and the potential mechanisms that link these two conditions are not clear.

METHODSA total of 179 Han Chinese subjects were enrolled in this study. All subjects underwent polysomnography, the oral glucose tolerance-insulin releasing test (OGTT-IRT) and serum HbA(1)c measurement. Indexes including homeostasis model assessment-IR (HOMA-IR), Matsuda index, HOMA-β, early phase insulinogenic index (ΔI(30)/ΔG(30)), AUC-I(180) and oral disposition index (DIo) were calculated for the assessment of insulin resistance and pancreatic β-cell function.

RESULTSBased on OGTT, 25.4%, 44.6% and 54.5% subjects were diagnosed having glucose metabolic disorders respectively in control, mild to moderate and severe OSAS groups (P < 0.05). Serum HbA(1)c levels were highest in subjects with severe OSAS (P < 0.05). In contrast, compared with normal subjects, HOMA-β, ΔI(30)/Δ(G30) and DIO were lower in severe OSAS group (P < 0.05). In stepwise multiple linear regressions, 0-min glucose and HbA(1)c were positively correlated with the percentage of total sleep time below an oxyhemoglobin saturation of 90% (T90) (Beta = 0.215 and 0.368, P < 0.05); 30-min and 60-min glucose was negatively correlated with the lowest SpOO(2) (LSpO(2)) (Beta = -0.214 and -0.241, P < 0.05). HOMA-β and DI(O) were negatively correlated with T90 (Beta = -0.153 and -0.169, P < 0.05) while body mass index (BMI) was the only determinant of HOMA-IR and Matsuda index.

CONCLUSIONSOSAS is associated with impairment in glucose tolerance and pancreatic β-cell function in Han Chinese subjects while insulin sensitivity is mainly determined by obesity.

Adolescent ; Adult ; Aged ; Female ; Glucose ; metabolism ; Glucose Tolerance Test ; Glycated Hemoglobin A ; analysis ; Humans ; Insulin Resistance ; Insulin-Secreting Cells ; physiology ; Male ; Middle Aged ; Polysomnography ; Sleep Apnea, Obstructive ; metabolism

BACKGROUNDIt has been shown that neurohumoral factors other than mechanical obstruction are involved in the pathophysiology of acute pulmonary thromboembolism (APTE). The aim of this study was to investigate the effects of thrombolytic drugs, a selective endothelin-1 receptor (ET-1R) antagonist alone or their combination on APTE in a canine model.

METHODSTwenty dogs were randomly assigned to five groups: sham, model, urokinase (UK), BQ123, and combination (UK plus BQ123). The dogs in the sham group underwent sham surgery. APTE was induced in the other four groups by intravenous injection of autologous blood clots. Dogs in the UK, BQ123 and combination groups received UK, BQ123 (a selective ET-1R antagonist), or UK plus BQ123, respectively. The dogs in the model group were given saline. Mean pulmonary artery pressure (mPAP), serum concentrations of ET-1, thromboxane (TXB2), and tumor necrosis factor (TNF)-alpha were determined at different time points following the induction of APTE.

RESULTSUK and BQ123 alone markedly decreased mPAP in APTE. By comparison, the reduction was more significant in the combination group. Compared with the sham group ((-0.90 +/- 0.61) mmHg), mPAP increased by (7.44 +/- 1.04), (3.42 +/- 1.12) and (1.14 +/- 0.55) mmHg in the model group, UK alone and BQ123 alone groups, respectively, and decreased by (2.24 +/- 0.67) mmHg in the combination group (P < 0.01). Serum ET-1 concentrations in the BQ123 and combination groups were (52.95 +/- 8.53) and (74.42 +/- 10.27) pg/ml, respectively, and were significantly lower than those in the model and UK groups ((84.56 +/- 7.44) and (97.66 +/- 8.31) pg/ml respectively; P < 0.01). Serum TNF-alpha concentrations were significantly lower in the BQ123 group than in the model, UK and combination groups (P < 0.05).

CONCLUSIONSOur results indicate that the selective ET-1R antagonist BQ123 not only reduces the increase of mPAP and serum ET-1 level, but also inhibits the production of TNF-alpha, and attenuates the local inflammatory response induced by APTE. Selective ET-1R antagonists may be beneficial to the treatment of APTE, particularly when used in combination with a thrombolytic agent.

Animals ; Dogs ; Endothelin A Receptor Antagonists ; Endothelin-1 ; blood ; Fibrinolytic Agents ; therapeutic use ; Peptides, Cyclic ; therapeutic use ; Pulmonary Embolism ; blood ; drug therapy ; pathology ; Random Allocation ; Thromboxanes ; blood ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo isolate viruses from mosquitoes in the south of Xinjiang and identify these viruses primarily.

METHODSA total of 13 491 mosquitoes were collected in the south of Xinjiang from Jul to Aug, 2005. These mosquitoes were divided into 130 groups and grinded respectively. The supernates were inoculated in C6/36 and Vero cells. Viruses isolated were detected, the genomic nucleic types by electrophoresis of viral genomes and the morphologies observed under electronmicroscope.

RESULTSAll 42 viruses were isolated, which caused CPEs on C6/36 but not on Vero cells. 27 viruses showed similar genomic profiles with 12 dsRNA segments. 1 virus displayed genomic profile with 10 dsRNA segments. 5 viruses took on similar genomic profiles with about 4 kbp DNA band. 9 viruses did not get any taxonomy information. Electromicroscopic pictures of these viruses revealed that above four types of viruses had distinguished morphologies indicating different virus species.

CONCLUSIONThere should be several virus species in the mosquitoes in the south of Xinjiang. dsRNA virus with 12 genomic segments should play analysis a predominant role in the south of Xinjiang.

Animals ; Bluetongue virus ; classification ; genetics ; isolation & purification ; Cercopithecus aethiops ; China ; Culicidae ; virology ; Dengue Virus ; classification ; genetics ; isolation & purification ; Genome, Viral ; Insect Viruses ; classification ; genetics ; isolation & purification ; RNA, Double-Stranded ; genetics ; RNA, Viral ; genetics ; Reassortant Viruses ; genetics ; Sequence Analysis, DNA ; Vero Cells

OBJECTIVETo probe the primary characteristic of 0507JS11 virus isolated from Culex sp. and determine the classification of 0507JS11 virus in taxonomy.

METHODS0507JS11 virus was cultured in Aedes albopictus C6/36 cells and cytopathic effects (CPEs) were recorded. Electro-microscopic morphology of 0507JS11 virus was observed. Total DNA extract of 0507JS11 virus was detected by 1% Agarose Gel Electrophoresis. Complete genomic sequence of 0507JS11 virus was sequenced and then made phylogenetic analysis.

RESULTS0507JS11 virus could cause CPEs in Aedes albopictus C6/36 cells. Viral particles have no envelope and appear icosahedron symmetry with diameter of 20 nm. The genome of 0507JS11 virus was positive single strand DNA (ssDNA) with full length of 3977 nt. However, a DNA band about 4 kbp was observed in the electrophoresis of total DNA extract of 0507JS11 virus. The coding region of the genome included three ORFs, ORF1 and ORF2 code NSP1 and NSP2, ORF3 codes VP. Phylogenetic analysis of the complete genomic sequence of 0507JS11 virus indicated an independent linear in Brevidensovirus.

CONCLUSION0507JS11 virus is a new member in Brevidensovirus.

Animals ; Culex ; virology ; DNA, Viral ; genetics ; Densovirinae ; classification ; genetics ; isolation & purification ; Genome, Viral ; Sequence Analysis, DNA