OBJECTIVETo establish a rapid specific method to identify the single-nucleotide polymorphisms (SNPs) of HBV polymerase gene region which are the methionine residue of the conserved YMDD motif.

METHODSTwo specific primers were designed to amplify interested gene region involved in SNPs which were also used as HBV DNA identification. Specific primers of SNaPshot were designed to detect 741A-G (YVDD), 743G-T (YIDD). The different fluorescent dye labeled ddNTP was used to further extend the strand of PCR product and was detected by ABI PRISM 310 Genetic Analyzer. Sera from 13 patients with chronic hepatitis B after lamivudine treatment were analyzed.

RESULTSAside from mutation of YMDD, there were mutations of 514C-A, 523C-A, 562T-A, 667C-A. The 13 samples were simultaneously tested with SNaPshot and DNA sequencing, the same results were obtained. The method of SNaPshot showed high specificity.

CONCLUSIONMutation of YMDD results in the changes of ATG codon, and there are new ATG codon in the upper strand of YMDD. SNaPshot technique is rapid, specific and accurate for the SNPs monitoring of HBV DNA mutation during lamivudine therapy. Two samples were determined by SnaPshot technique, identifying the co-existence of the mixed wild type and mutant type HBV infection.

Antiviral Agents ; therapeutic use ; Base Sequence ; DNA, Viral ; blood ; genetics ; Drug Resistance, Viral ; Gene Products, pol ; genetics ; Hepatitis B ; blood ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVETo compare the efficacy of nasal synchronized intermittent positive pressure ventilation (NSIPPV) and nasal continuous positive airway pressure ventilation (NCPAP) for the treatment of apnea in preterm infants.

METHODSEighty preterm infants with apnea from August 2010 to January 2011 were randomly administered with NSIPPV and NCPAP (n=40 each).The blood gas results before and 2 hrs after ventilation, time of using ventilator, therapeutic efficacy and complications were compared between the two groups.

RESULTSThere were no significant differences in the blood gas results between the two groups before ventilation. The blood gas results (pH, PO2, PCO2) in the NSIPPV group were better than those in the NCPAP group 2 hrs after ventilation. The time of using ventilator in the NSIPPV group was shorter than that in the NCPAP group (50±9 h vs 91±11 h; P<0.01). There were no significant differences in the total effective rate between the NSIPPV and the NCPAP groups (95% vs 85%; P>0.05). The proportion of ventilator weaning within 3 days in the NSIPPV group (23/40) was higher than that in the NCPAP group (14/40) (P<0.05). The incidence of complications in the NSIPPV group was not different from that in the NCPAP group (22% vs 25%).

CONCLUSIONSNSIPPV appears to be superior to NCPAP for the treatment of apnea in preterm infants.

Apnea ; therapy ; Blood Gas Analysis ; Continuous Positive Airway Pressure ; Humans ; Infant, Newborn ; Infant, Premature ; Intermittent Positive-Pressure Ventilation ; adverse effects ; methods

OBJECTIVETo study the efficacy of endotracheal lavage in neonatal ventilator-associated pneumonia (VAP).

METHODSFifty-eight neonates with VAP between January 2002 and December 2008 were randomly assigned to two groups: lavage and control (n=29 each). After withdrawal from ventilator, both groups received sensitive antibiotics therapy according to sputum culture results as well as supportive treatment. The lavage group was additionally treated with endotracheal lavage (2-3 times daily). The therapeutic effects were compared between the two groups.

RESULTSThere were no significant differences in the average time of mechanical ventilation between the lavage and the control groups. The effective rate in the lavage group (93%) was significantly higher than that in the control group (69%; p<0.05). Three percent of patients in the lavage group required twice or more mechanical ventilation compared with 24% in the control group (p<0.05). Blood gas analysis results were obviously improved in the lavage group 2 hrs after treatment (p<0.01).

CONCLUSIONSEndotracheal lavage can decrease the number in mechanical ventilation and improve therapeutic effects in neonates with VAP.

Female ; Humans ; Incidence ; Infant, Newborn ; Male ; Pneumonia, Ventilator-Associated ; epidemiology ; therapy ; Respiration, Artificial ; statistics & numerical data ; Therapeutic Irrigation ; methods ; Trachea

Both phytase and endoglucanase are additives in feed for mono-gastric animal known for their effects. Recombinant vector pPICZalpha-EG was constructed and transformed to GS115-phyA, a Pichia pastoris strain that had integrated with phytase gene, generating GS115-phyA-EG. Both phytase and endoglucanase activities in the supernatant were determined after methanol induction of GS115-phyA-EG. Phytase and endoglucanase activity reached 39.4% and 56.2% activity compared to GS115-phyA and GS115-EG, respectively. Properties of the mixed enzyme suggest that the optimal temperature and pH value be 55 degrees C and 5.5 respectively. Both phytase and endoglucanase showed greater than 80% activity across temperature ranges 45 degrees C to 55 degrees C and pH ranges 4.5 to 5.5. Expressing more than one enzyme in one system could save time and money during induced expression, and the mixed enzyme might apply for treating forge before feeding with poultry.
6-Phytase ; biosynthesis ; genetics ; Cellulase ; biosynthesis ; genetics ; Genetic Vectors ; Pichia ; enzymology ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

To observe a PPAR-alpha agonist effect of N-oleoylethanolamine (OEA) on CB2 (cannabinoid receptor 2), an anti-inflammatory receptor in vascular endothelial cell, healthy HUVECs and TNF-alpha induced HUVECs were used to establish a human vascular endothelial cell inflammatory model. Different doses of OEA (10, 50 and 100 micromol x L(-1)) had been given to HUVECs, cultured at 37 degrees C for 7 h and then collected the total protein and total mRNA. CB2 protein expression was detected by Western blotting and CB2 mRNA expression was assayed by real-time PCR. As the results shown, OEA (10 and 50 micromol x L(-1)) could induce the CB2 protein and mRNA expression, but not 100 micromol x L(-1). To detect if anti-inflammation effect of OEA is partly through CB2, CB2 inhibitor AM630 was used to inhibit HUVEC CB2 expression, then the VCAM-1 expression induced by TNF-alpha was detected, or THP-1 adhere to TNF-alpha induced HUVECs was examined. OEA (50 micromol x L(-1)) could inhibit TNF-alpha induced VCAM-1 expression and THP-1 adhere to HUVECs, these effects could be partly inhibited by a CB2 inhibitor AM630. The anti-inflammation effect of OEA is induced by PPAR-alpha and CB2, suggesting that CB2 signaling could be a target for anti-atherosclerosis, OEA have wide effect in anti-inflammation, it may have better therapeutic potential in anti-inflammation in HUVECs, thus achieving anti-atherosclerosis effect.
Anti-Inflammatory Agents ; pharmacology ; Atherosclerosis ; pathology ; Cell Adhesion ; drug effects ; Cells, Cultured ; Endocannabinoids ; pharmacology ; Endothelial Cells ; cytology ; metabolism ; Ethanolamines ; pharmacology ; Humans ; Indoles ; pharmacology ; Monocytes ; drug effects ; Oleic Acids ; pharmacology ; PPAR alpha ; antagonists & inhibitors ; RNA, Messenger ; metabolism ; Receptor, Cannabinoid, CB2 ; antagonists & inhibitors ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo evaluate the clinical effect of endotracheal lavage with porcine pulmonary surfactant (PS) in term neonates with severe meconium aspiration syndrome (MAS).

METHODSA total of 136 full-term infants with severe MAS who were admitted to the neonatal intensive care unit between January 2010 and June 2013 were randomly and equally divided into PS lavage and PS injection groups. In the PS lavage group, patients were treated with endotracheal lavage using 3-5 mL of diluted PS (12 mg/mL) each time, and the PS injection group was given PS by intratracheal injection at the first dose of 200 mg/kg. Blood gas, oxygenation index (OI), and PaO2/FiO2 (P/F) of the two groups were evaluated before and 2, 12, 24, and 48 hours after the treatment, and the duration of mechanical ventilation, complication rate, and cure rate were compared between the two groups.

RESULTSCompared with the PS injection group, the PS lavage group had significantly higher PaO2 and P/F ration and significantly lower PaCO2 and OI at 12, 24, and 48 hours post-treatment (P<0.01), a significantly shorter duration of mechanical ventilation (P<0.01), a significantly smaller amount of PS (P<0.01), a significantly lower complication rate (P<0.05), and a significantly higher cure rate (97% vs 88%; P<0.05).

CONCLUSIONSCompared with the intratracheal injection of PS, endotracheal lavage with diluted PS in term neonates with severe MAS can increase ventilation and oxygenation efficiency, shorten the duration of mechanical ventilation, reduce the complication rate, and increase the cure rate, indicating that this method is a safe and effective therapeutic strategy.

Animals ; Humans ; Infant, Newborn ; Meconium Aspiration Syndrome ; drug therapy ; Pulmonary Surfactants ; administration & dosage ; Swine ; Therapeutic Irrigation ; Trachea

Objective To explore the Electrocardiography (ECG) changes of residents in Keshan disease area and the status of growth and decline of Keshan disease in Shaanxi Province. Methods Using stratified randomized sampling method,2 mild,2 moderate and 2 serious disease counties were selected respectively in 2005 and 2006. A total of 6 counties were sampled,2 villages,one with severe disease and one with mild,were selected from each sampled county. A total of 12 villages were selected. The clinical examination and ECG were conducted in 3-year old children of agricultural population of the selected villages. Results ECG of 5692 cases were performed in the selected 12 village in the 6 counties,in which 4917 cases showed normal electrocardiogram,up to 86.38% (4917/5692). Two hundred and fifty-two cases showed roughly normal electrocardiograms,up to 4.43%(252/5692). Five hundred and twenty-three cases had abnormal electrocardiogram,accounting for 9.19% (523/5692). Among them,the abnormal electrocardiogram rates in mild,moderate and serious disease areas were 7.07% (144/2036), 11.41%(167/1646) and 10.54%(212/2010),respectively. Atrioventricular block was the major abnormal electrocardiogram change,followed by arrhythmia,ST-T changes,and low voltage. One hundred and thirty-nine cases were confirmed as latent and chronic Keshan diseases. One hundred and thirty-one cases were latent Keshan, and detection rate was 2.30%(131/5692). Eight cases were chronic Keshan,and the detective rate was 0.14% (8/5692). Complete right bundle branch block [37.06% (63/170) ],ST-T changes [22.35% (38/170) ],multiple premature ventricular beats [12.94% (22/170)] were the major abnormal electrocardiogram change of Keshan patients. Conclusions Atrioventricular block and arrhythmia are the major abnormal electrocardiogram changes. Keshan disease incidences are controlled under a stable condition.

Objective To know the prevalence tendency of Keshan disease(KSD) under control after 10 years in Shaanxi Province, the factors that causes or relative to the disease, to provide scientific reference for disease's prevention and control. Methods Through stratified cluster sampling, based on the severity of KSD in endemic area of Shaanxi Province, 12 villages from 6 counties were randomly selected as investigation points in 2006. The people older than 3 year-old were chosen to do clinical check up and electrocardiogram tracing. Among them, suspicious or abnormal cases were asked to take chest X-ray and cardiac ultrasound. Maize and rice, hair and whole blood were randomly collected to test the selenium content, the activity of Glutathione peroxidase (GSH-Px). Results The total detection rate of potential or chronic KSD was 2.44%(139/5694), the detection rate of abnormal ECG was 9.19% (523/5692), the detection rate of cardiac enlargement from chest X-ray and cardiac ultrasound were 45.6%(72/158) and 34.5%(59/171) respectively. The average content of selenium in staple foods(wheat and corn) were[(0.045±0.036), (0.035±0.025)mg/kg, respectively]. The level of hair selenium in patients and healthy people were [(0.376±0.091), (0.384±0.077)mg/kg, respectively], with non-significant different (u=0.77, P>0.05). There were significant differences in whole blood selenium of patients, healthy people in KSD areas and healthy people in non-KSD areas[(0.071±0.017), (0.077±0.017), (0.090±0.016)mg/L, respectively; F=4.55, P<0.05), the whole blood selenium in patients lower than in healthy people in KSD areas (P<0.05), in healthy people in KSD areas lower than in non-KSD areas (P<0.05). Conclusions After the KSD condition being controlled, the situation in Shaanxi Province has become stable and exhibited a decreasing tendency. The selenium level of both internal and external environment in the endemic area increased significantly, that is the main factors of controlling disease.

Objective To explore the status of Keshan disease in Shaanxi province to provide a scientific basis for decision-making of prevention and control of Keshan disease. Methods Nineteen infected villages were randomly selected in 19 infected counties in the range of Keshan disease infected area in Shaanxi province in 2008 as the investigation sites. Clinical examination and electrocardiography were performed in the chosen people at every spots, chest X-ray of posteroanterior position film in 2-meter distance was taken in suspicious cardiac patients, and determining the selenium contents was also determined in the collected grain samples of the investigators. Results Of the 10 228 investigated residents in the endemic area, 110 Keshan disease patients were detected, the total detection rate was 1.08% (110/10 228). Among the 110 patients, 92 were potential Keshan disease, which accounted 0.90%(92/10 228); 18 chronic Keshan disease formed a detection rate of 0.18%( 18/10 228); no acute and sub-acute type of Keshan disease had been inspected. Potential Keshan disease patients often showed electrocardiogram abnormality of complete fight bundle branch block [48.57%(51/105)], ST-T change[ 19.05% ( 20/105 ) ], frequent premature ventricular contraction [ 10.48 % ( 11/105 ) ], left ventricular hypertrophy [ 5.71% (6/105) ], block in the anterosuperior division of the left branch[5.71%(6/105)]; Chronic of Keshan patients mostly presented atrial fibrillation [ 24.00% (6/25) ], left ventricular hypertrophy [ 20.00% (5/25) ], complete right bundle branch block [ 20.00% (5/25)]. The increase rate of cardiothoracie ratio was 18.08% (32/177). Food samples of wheat, corn, millet and rice in infected area residents were of selenium content, being (0.096± 0.028), (0.089 ±0.029), (0.087 ± 0.016), (0.047 ± 0.016)mg/kg, respectively. Conclusions Keshan disease in Shaanxi province is steadily declining, potential and chronic Keshan diseases are currently the main clinical types. Selenium content of food in endemic area has reached the level of the non-endemic area.

OBJECTIVETo investigate the ways testosterone influences the murine bone marrow-derived macrophages (BMMs) and how testosterone affects the function of BMMs after bound to their membrane surface.

METHODSBMMs were cultured in vitro, their total RNA and proteins isolated, and the expression of intracellular androgen receptor (AR) detected through RT-PCR and Western blotting. The binding site of testosterone (T) to the membrane surface of BMMs was observed by confocal laser scanning microscopy after T-BSA-FITC incubation. Moreover, the intracellular Ca2+ was tested by Fura-2 method, and the influence of ionic currents on BMMs plasma membrane induced by testosterone was examined by the whole cell patch-clamp.

RESULTSRT-PCR and Western blotting failed to detect intracellular ARs in BMMs, but confocal laser scanning microscopy showed testosterone to be bound to the membrane surface of BMMs by impermeable T-BSA-FITC, inducing a rapid rise in the intracellular free Ca2+ concentration ([Ca2+]i) of Fura-2 loaded BMMs, predominantly due to the influx of extracellular Ca2+ through Ni2+ -blockable Ca2+ channels in the plasma membrane. Similarly, the patch-clamp technique revealed T-induced calcium influx in BMMs.

CONCLUSIONIt is reasonable to assume that the testosterone receptor exists on the plasma membranes, and testosterone act through unconventional plasma membrane receptors, induce Ca2+ influx and a rapid rise in the intracellular Ca2+ concentration, and influence the function of BMMs.

Animals ; Blotting, Western ; Calcium ; metabolism ; Calcium Channels ; physiology ; Cell Membrane ; metabolism ; Cells, Cultured ; Female ; Macrophages ; cytology ; metabolism ; physiology ; Membrane Potentials ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Protein Binding ; Receptors, Androgen ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; metabolism