In this paper, vegetative( vip83 ) and crystal(cry1Ac10 and cry1Ca) insecticidal protein genes from Bacillus thuri ngiensis were simultaneously electrospored into the plasmid-free strain BMB17 1. By the means of the specific P CR detection, the recombinant strains BMB2830-171 contained cry 1Ac10 and vip83, and BMB2 882-171 had cry1Ca and vip83 , were obtained respectively. Under the control of r ecombinant strains with one gene, bioassay of the synergism between vegetative V ip83 and crystal Cry1Ac10( or Cry1Ca )insecticidal proteins to three important Lepidopteran pests were done. The results, by analysis of statistic bio-so ft, showed that the synergia relation of vegetative Vip83 and crystal Cry1Ac10 i nsecticidal protein toxic to Heliothis armigera wascounteracted, while Plu tella xylostella and Spodotera exigua unobservable. There was no synergis tic action between Vip83 and Cry1Ca insecticidal proteins with Spodotera exigu a as tested insect. Bu t their cooperation to Heliothis armigera was minus, and the counterpart to Plutella xylostella plus, whose cotoxicity factor is 32.6. The experiment of bi-g ene genetic stability also suggested that the synergia effection had certain molecu lar genetic stability in the same cell. This performance can be contributed to construct high-effect and wide-spectrum engineered strain.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Diseases ; diagnosis ; Bile Duct Neoplasms ; diagnosis ; Cholangiopancreatography, Endoscopic Retrograde ; Cytodiagnosis ; Cytological Techniques ; methods ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Diseases ; diagnosis ; Pancreatic Neoplasms ; diagnosis

Bone Density ; Bone and Bones ; metabolism ; Hepatitis B ; complications ; Humans ; Liver Cirrhosis ; metabolism ; Male ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVELong time exhaled oxygen will induced oxygen toxicity. Some studies had found that different pathology may exised in normobaric and hyperbaric pulmonary oxygen toxicity, and nitric oxide synthase (NOS) may play a role. In this study, we discussed the change of NOS in normobaric and hyperbaric pulmonary oxygen toxicity.

METHODSSixty male SD rats were randomly divided into 6 groups (n = 10), exposed to 1 ATA (atmosphere absolute), 1.5 ATA, 2 ATA, 2.5 ATA and 3 ATA, 100% oxygen for 56, 20, 10, 8, 6 hours respectively. Rats were exposed to air as control. After exposure, the protein in bronchoalveolar lavage fluid (BALF), the wet/dry weight of lung and the expression of eNOS, nNOS in lung were defined.

RESULTSAs compared to air group, the protein in BALF, the wet/dry of lung were significantly elevated in 1.0 ATA group, while these changes were not so obviously in the other groups, and these changes in hyperbaric oxygen group (approximately 1.0 ATA) were significantly decreased as compared with nonnrmobaric oxygen group (1.0 ATA). The expression of nNOS were not changed in normobaric and hyperbaric pulmonary oxygen toxicity, while the expression of eNOS was significantly decreased in 2 ATA group, and significantly elevated in 2.5 ATA and 3 ATA group.

CONCLUSIONThe expression of eNOS can change when exposed to different pressures of oxygen.

Animals ; Disease Models, Animal ; Lung ; metabolism ; Male ; Nitric Oxide Synthase Type I ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxygen ; poisoning ; Pressure ; Rats ; Rats, Sprague-Dawley

The aim of the research is to investigate the genetic characteristics of Legionella pneumophila serogroup1 (LP1)in Sichuan Province.The sequence-based typing (SBT) and multiple-locus VNTR analysis (MLVA) were used to describe the genetic polymorphism of 42 strains which were isolated from 1989-2016 in Sichuan Province,China.According to the reference,PCR was used to detect the 8-VNTR loci and 7 housekeeping genes respectively.The VNTR results were determined by using capillary electrophoresis,and the SBT results were sequenced and compared with the database of EWGLI.Results showed that totally 42 stains were divided into 8 MLVA types with the advantage types were M08 (47.6 ％) and M07 (23.8 ％).Twelve ST types were obtained with 3 main clonal complex and 2 singleton,including 2 novel ST types,among those,ST1 was the predominant type,accounting for 52.3 ％,following by ST630 (14.2 ％).In conclusion,our results demonstrated MLVA and SBT were both applied to the research for molecular epidemiological investigation of LP1 and showed the high genetic polymorphism and the regional specificity.The results also suggest that the isolates are a potential threat to the public,effective control and prevention strategies are urgently needed.

Objective: To investigate the application of gradient cushion on prevention of supine hypotension syndrome (SHS) undergoing cesarean section. Methods: 450 parturients undergoing cesarean section with spinal and epidural anesthesia, aged 20-45 years, ASA Ⅰ, Ⅱor Ⅲ grades, were randomly assigned into three groups: gradient cushion group (group A), sandbag group (group B) and left-leaning-operating table group (group C), 150 cases in each. The posture intervention was alternated after completion of spinal and epidural anesthesia. Recorded the cases of SHS, and collected systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), respiratory rate (RR) and saturation of pulse oximetry (SpO₂) before anesthesia, 2 min, 5 min, 10 min after anesthesia and prefetus removal from uterus. And assessed the position comfort with surgical posture comfort scale. Results: The incidence of SHS in group A was 8.0%(12/150), in group B was 20.0% (30/150), and in group C was 21.3% (32/150). The rate of SHS was higher in group A than other groups (χ² value was 8.970, 10.653, all P<0.01). The score with surgical posture comfort scale was (47.03 ± 3.01), (38.13 ± 4.70), (36.10 ± 4.04), which was higher in group A than group B or group C, and the score with surgical posture comfort scale was higher in group B than group C (t value was 27.413, 30.227, 2.542, P<0.01 or 0.05). SBP and DBP were higher in group A and group B than group C at 2 min, 5 min, 10 min after anesthesia and prefetus removal from uterus, and SBP and DBP were higher in group A than group B at 2 min, 5 min, 10 min after anesthesia and prefetus removal from uterus, HR and RR were higher in group A and group B than group C at 2 min, 5 min, 10 min after anesthesia and prefetus removal from uterus, and HR and RR were higher in group A than group B at 2 min, 5 min, 10 min after anesthesia and prefetus removal from uterus (t value was -15.842-21.117, P<0.05 or 0.01). Conclusion After spinal and epidural anesthesia, applying the gradient cushion for adjustment of position would be effective to reduce the occurrence of SHS, simple to handle, decreasing to change the position and increasing to comfort after position in cesarean section.

Objective To evaluate the self-treatment effectiveness on patients with ehwnic Keshan disease.Methods Twenty patients with chronic Keshan disease were selected from individuals with Keshan disease in Fuyu County,Heilongjiang Province.They were trained three times every three months of self management including pathogenetic condition education,general guidance,drug therapy,and they also taught how to adiust the doBe of drug according to their illness.Major symptom score,heart rate(HR),ultrasoundcardiogram (UCG)index and cardiac functional grading of these patients at basehne,after 3 months and 6 months of treatment were compared.Results The 20 patients rated their main symptoms score as(15.03 ±6.77)before self- treatrnent,and significantly decreased to(7.25±4.82)and(6.70±4.90)after 3 and 6 months treatment(P<0.01); the heart rate(HR) was (76.40±12.06) beats per minute(bpm)before self-treatment,and dramatically decreased to (69.95±12.63),(67.15±9.76)bpm after 3 and 6 months treatment(P<0.01).As for UCG detecting index,left atrial diameter(Lad)aIld left ventricular end-diastolic diameter(LVEDd)Was(37.85 ±5.23)nun and(52.49± 9.38)mm separately before self-treatment,and notablely decreased to(36.77 ±5.63),(52.15 ±9.24)mm,and (35.29±5.50),(50.81±8.88)mm respectirely after 3 and 6 months of treatment(P<0.01 or<0.05);left ventricuIar ejection fractiOII(LVEF)markedly increased(P<0.05),from(55.15±15.80)%at baseline to(57.35± 12.51)%at 3 months and(60.30±13.42)%at 6 months;there were no significant differences in mitral flow E/A ratio changes before and after treatment(P>0.05);compared with prior to the treatment.cardiac function grading was significantly better aftertreatmentfor 3 months(T=36.0,P<0.05),but not after 6 months(T=17.5,P> 0.05).Conclusions The patients'serf-treatment is effective,which we recommend to uphold and widespread.

Objective To observe the change in cardiac shape and heart function and evaluate the effect of self-treatment on patients with Keshan disease by echocardiography. Methods To check the 31 patients with Keshan disease before the self-treatment, and follow them up in the 3rd and 6th months after self-treatment by echocardiography. The left atrium diameter(LAd), left ventricular end-diastolic diameter(LVEDd), the thickness of interventricular septum in end-diastolic(IVSTd), the thickness of LV posterior wall in end-diastolic (LVPWTd), left ventricular mass(LVM), left ventricular mass index(LVMI), left ventricular ejection fraction(LVEF) and mitral valve flow E/A ratio(E/A) were measured. Results The LAd[(35.8±5.1)ram] and LVPWTd[(9.3±1.0)mm] obviously decreased in the 3rd month after serf-treatment compared with prior self-treatment [ (37.0±5.0), (9.9± 1.2)mm](P<0.05). The LAd[(34.5±5.0)mini, IVSTd[(9.5±1.3)mm], LVEDd[(50.2±7.7)mm], LVPWTd [(8.7±1.1)mm], LVM[(196.1±87.2)g] and LVMl[(126.5±56.4)g/m2] obviously decreased in the 6th month after self-treatment compared with prior self-treatment [(37.0±5.0), (10.2±1.5), (51.3±8.1), (9.9±1.2)mm, (230.4±95.5)g, (144.0±54.6)g/m2] and in the 3rd month after self-treatment [(35.8±5.1)mm, (10.2±1.4) ram, (51.1±8.1)nun, (9.3±1.0)mm, (219.4±82.5)g, (136.8±50.0)g/m2] (P<0.05). The results of the mitral valve flow E/A ratio and LVEF in the 3nt month after self-treatment [1.0±0.5, (59.4±13.3)%] were increased compared with the prior self-treatment[0.9±0.5, (58.1±15.6)%], and the results in the 6th month after self-treat- ment[ 1.0±0.4, (60.7±13.6)%] were further inereased compared with before, but there was no signifieant differ- ence(P0.05). Conclusions Self-treatment of Keshan disease patients can improve the heart function by pre- venting left ventrieular remodeling and reversing. Echocardiography can be used as an essential technique to evalu- ate the effect of self-treatment on Keshan disease patients.

Three kinds of Bacillus thuringiensis serotype-subsp. Leesis(H33) strain YBT-833, subsp. Aizawai(H7) strain YBT-1416 and subsp. Kurstaki(H3ab) strain YBT-1535, which were isolated by our lab, are chosen as original strain to clone vegetative insecticidal protein gene. Southern hybridization showed that vip genes are all localized at roughly 4-5 kb size-fractionated XbaI fragments of total DNA from YBT-833, YBT-1416 and YBT-1535. Three subgenomic libraries containing the vip gene fragment, were constructed with pUC19 as vector. Then, three vegetative insecticidal protein gene vip83, vip14 and vip15 are obtained from the libraries through the methods of colony-blot-in-situ screening and enzyme-cut detection. Comparision of DNA sequence made out that only vip83 gene exist five different base pairs with known vip genes. Because the sequences of vip14 and vip15 are the same, two of the three genes, vip83 and vip14, were subcloned to shuttle vehicle pHT315 to get recombinant plasmids pBMB8901 and pBMB8902 in turn. The plasmids were separately transformed into vip Bt. receptors BMB171 and 4Q7 to obtain four engineered strains BMB8901-171, BMB8902-171, BMB8901-4Q7 and BMB8902-4Q7. SDS-PAGE results indicated that all recombinant strains express 88 kD vegetative insecticidal protein. Bioassay also showed that the proteins of genes vip83 and vip14 both have certain toxicity to Lepidopteran insect larvae such as Heliochis armigera, Spodotera exigua and Plutella xylostella. While the toxicity of vip protein from four engineered strains to Plutella xylostellas are highest, whose LC50 value is 28.6, 31.6, 45.4 and 37.6 microL/mL respectively. This study will contributed to construct high efficacy and wide spectrum engineered strains on theory and reality.
Animals ; Bacillus thuringiensis ; genetics ; Bacterial Proteins ; chemistry ; genetics ; pharmacology ; Cloning, Molecular ; Insecticides ; pharmacology ; Pest Control, Biological ; Recombinant Proteins ; biosynthesis ; pharmacology

OBJECTIVETo study the pathogenesis of abnormal bone metabolism in patients with HBV liver cirrhosis.

METHODSNM-300 signal-energy X-ray absorptiometry system was used to measure the bone mineral density (BMD) in 61 liver cirrhosis patients and 30 age-matched healthy controls. The serum levels of 1,25(OH)2D3, parathyroid hormone (PTH), calcitonin (CT), bone gamma-carboxyglutamic acid-containing protein (BGP), IL-1beta, IL-6, tumor necrosis factor (TNF)alpha and urine crosslaps were also detected in these patients.

RESULTSBMD in patients with HBV liver cirrhosis was lower than those of the controls. The serum levels of 1,25(OH)2D3 and BGP in cirrhosis patients were lower than those in the controls, and they were much lower in the osteoporosis (OP) group than in the non-osteoporosis (NOP) group. The PTH and CT were higher significantly in the patients than in the controls. The changes of serum 1,25(OH)2D3 and BGP were correlated with the changes of BMD. The serum levels of IL-1beta, IL-6, TNFalpha and urine crosslaps in cirrhosis patients were higher than those of the controls, and they were much higher in the OP group than in the NOP group. We also found that the serum levels of IL-1beta, IL-6, TNFalpha and urine crosslaps had a negative correlation with BMD.

CONCLUSIONSThese data suggest that bone formation is weakened and bone resorption is increased in patients with HBV liver cirrhosis, 1,25(OH)2D3 plays an important role in abnormal bone formation. Elevation of serum IL-1beta, IL-6, TNFalpha can accelerate bone resorption and cause hepatic bone disease (HBD). Taking 1,25(OH)2D3 and reducing the level of IL-1beta, IL-6, TNFalpha may be very important in preventing and treating HBD.

Adult ; Bone Density ; Bone and Bones ; metabolism ; Calcitriol ; pharmacology ; Hepatitis B, Chronic ; complications ; Humans ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Osteoporosis ; etiology