To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.

The research aimed to investigate the therapeutic effects and mechanisms of Opuntia dillenii Haw polysaccharide (OPS) on atherosclerosis of rats. First atherosclerotic rat models were established by high-fat and high-calcium diet. Thirty days later, the rats were treated with low dosage of OPS (0.2 g x kg(-1) x d(-1)) or high dosage of OPS (0.4 g x kg(-1) x d(-1)) by intraperitoneal injection for 60 days continuously. At the end of treatment, thoracic aorta rings were prepared and vasorelaxation of rat thoracic aorta in different experiment groups were determined by using 620M multi wire myograph system in vitro. Blood and livers of rats were collected. Then plasma levels of total cholesterol (TC), triglyceride (TG) and low density lipoprotein (LDL) of rats were separately determined using whole automatic biochemical analyzer; protein level of hepatic apolipoprotein B (ApoB) and that of hepatic diglyceride acyltransferase (Dgat1) were measured by Western Blot technique. Results showed that the ability of rat thoracic aorta to relax decreased markedly in the model group compared with that in the normal group, and significant differences existed in vasorelaxation ratios induced by different concentrations of carbamylcholine chloride (Carb) between these two groups (P < 0.01). After OPS treatment, the ability of rat thoracic aorta to relax improved markedly, the vasorelaxation ratios induced by Carb at 5 and 10 μmol x L(-1) were respectively 0.34 ± 0.08 and 0.62 ± 0.15 in the group treated with low dosage of OPS, while the ratios induced by Carb at 1 and 5 μmol x L(-1) were respectively 0.54 ± 0.08 and 0.98 ± 0.02 in the group treated with high dosage of OPS, which were all significantly different with those in the model group (P < 0.01). Plasma contents of TC, TG and LDL reduced significantly by the treatments both with low and high dosages of OPS compared with those in the model group (P < 0.01). Protein level of hepatic ApoB and that of hepatic Dgat1 decreased significantly after the treatment with high dosage of OPS compared with those in the model group (P < 0.01). These results indicate that OPS can markedly improve the vasorelaxation of thoracic aorta of atherosclerotic rats and has significant anti-atherosclerotic effect; inhibiting the expression of ApoB and Dgat1 and thus decreasing the amounts of TC, LDL and TG serving as one of the molecular mechanisms of its antiatherosclerosis effect.
Animals ; Aorta, Thoracic ; drug effects ; Atherosclerosis ; drug therapy ; Cholesterol ; blood ; Lipoproteins, LDL ; blood ; Opuntia ; chemistry ; Phytotherapy ; Rats ; Triglycerides ; blood

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 purpose of this study was to establish a model of trigeminal neuralgia (TN) through an approach from lower edge of cheekbone and to observe the functional changes in the voltage-gated potassium currents in the cultured trigeminal ganglion (TG) neurons. Thirty Sprague-Dawley male rats were divided into two groups, the sham-operated (sham) group and the operated group. The TN model was carried out by using a chronic constriction injury of the infraorbital nerve (ION-CCI) from lower edge of cheekbone. Peripheral pain threshold test and whole-cell patch clamp recording were used to determine the difference between sham and ION-CCI rats. The withdrawal threshold of whisker pad in operated side of ION-CCI rat was decreased significantly from 6 d after operation and then maintained until 21 d, with the lowest on the 15th day. The threshold of whisker pad in non-operated side of operated rats was also decreased significantly compared with that in the sham group. Delayed rectifier potassium current (I(K)) in cultured ION-CCI TG neurons was decreased significantly compared with that in the sham group. Transient outward potassium currents (I(A)) in both operated and non-operated sides of TG neurons from ION-CCI rats were also reduced significantly compared with that in the sham group. The present study provided a new method of ION-CCI. In this model, the decrease of I(A) and I(K) might contribute, at least in part, to the decrease in mechanical pain threshold of whisker pad and the subsequent hyperalgia.
Animals ; Cells, Cultured ; Constriction ; Disease Models, Animal ; Hyperalgesia ; Male ; Pain Threshold ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; Rats ; Rats, Sprague-Dawley ; Trigeminal Ganglion ; metabolism ; Trigeminal Neuralgia ; physiopathology ; Vibrissae

To investigate the resources of medicinal plant, such as wild Apocynum, supervised classification based on Principal Component Analysis (PCA) and texture feature were used to monitor wild medicinal plants from image captured by ZY-3 and World-view-2 and compare which satellite Image are more appropriate to monitor the wild medicinal plants. The research results shows that: for more complex growth conditions wild medicinal plants Apocynum, high-resolution images Worldview-2 is more suitable for its remote identification, the low-resolution satellite ZY-3 can only recognizes the wild medicinal plants which distributed intensively. If the study target distribution is more intensive and larger scale, and cultivated type medicinal plants, the use of satellite ZY-3 in low resolution remote sensing data to identify the target can be a good choice, it is not necessary to buy high-resolution data, in order to avoid waste of expenditure, for the scattered distribution, the high-resolution satellite imagery data may be indispensable to identify targets.
Apocynum ; chemistry ; growth & development ; China ; Conservation of Natural Resources ; Geographic Information Systems ; Plant Dispersal ; Plants, Medicinal ; chemistry ; growth & development ; Remote Sensing Technology ; methods

Mycoplasma hyopneumoniae (M .hyopneumoniae) and Mycoplasma hyorhinis (M .hyorhinis) infections are common in China .To investigate the prevalence of M .hyorhinis and M .hyopneumoniae in Jiangsu Province of China ,a mo-lecular epidemiological survey was conducted from 399 nasal swab samples of unvaccinated pigs using nested Polymerase Chain Reaction (nested PCR) .Nasal swab samples were collected from Jiangquhai porcine lean (JQHPL) strain pigs and other West-ern breeds .Clinical samples were taken from each pig and divided into different groups based on ages of pigs (7 ,14 ,21 ,28 , and 35 days) .Results indicated that the prevalence of M .hyorhinis was 70 .9% from different herds in Jiangsu Province in China ,while the prevalence of M .hyopneumoniae was 13 .5% .M .hyorhinis infection was more common in pigs for less than 5 weeks of age compared to M .hyopneumoniae infection .Co-infection was also observed in 30 samples (7 .5% ) in which both M .hyorhinis and M .hyopneumoniae were detected .The M .hyorhinis infection increased as the animals grew from 7 to 35 days .The M .hyopneumoniae infection did not change significantly as the pigs grew older .Significant difference of M .hyorhi-nis infection was observed between other Western breeds and JQHPL pigs (P<0 .001) .JQHPL pigs appear to be more sensi-tive to the M .hyorhinis infection as compared to the other Western breeds .However ,there is no obvious relationship between the breed type and M .hyopneumoniae infection (P>0 .05) .

Objective To understand epidemiological characteristics of chronic obstructive pulmonary disease(COPD)in people aged over 40 years in a rural area of Tianjin.Methods Using cluster sampling,1 508 subjects over 40 years old at five villages in Xinkaikou Township,Baodi District,Tianjin were investigated with respiratory questionnaire,lung function test and physical examination.Confirmed patients with COPD were examined by chest roentgenography and electrocardiography.Results One hundred and forty-two subjects in that area suffered from COPD,with prevalence of 9.4%,24 of them (16.9%)were diagnosed as cor pulmonale.Prevalence of COPD increased with age,higher in men (13.5%)than that in women(6.2%),higher in smokers(12.2%)than that in non-smokers(7.2%), higher in those with family history(21.4%)than that in those without it(8.45%),and higher in those with coughing history during their childhood(75.0%)than that in those without it(9.2%),all with a P-value of less than 0.01.Univariate analysis showed that out-door air pollution,cooking,time length of burning firewood during cooking,smoking,coughing history during childhood,gender,age,family history all were predisposing factors for COPD.Multivariate analysis with logistic regression model showed that gender,age, family history were independently predisposing factors for COPD.Quality of life was better in non-COPD subjects than in those with COPD,with statistically significant difference.Conclusions Prevalence of COPD was relatively higher in people of rural Tianjin,with gender,age,family history and outdoor air pollution as main risk factors.

Objective To evaluate the clinical efficacy and safety of Shenfu injection in the treatment of leukopenia induced by chemotherapy docetaxel, epirubicin and cyclophosphamide adjuvant chemotherapy ( TEC chemotherapy) after surgery of breast cancer.Methods A total of 106 patients with breast cancer were randomly divided into control group ( n=53 ) and treatment group ( n =53 ) .Control group was re-ceived 75 mg? m-2 docetaxel, intravenous infusion on day 1 +50 mg? m-2 epirubicin, intravenous infusion on day 1 +500 mg? m-2 cy-clophosphamide, intravenous infusion on day 1, 3 weeks for a course. Treatment group was given 50 mL Shenfu injection added 500 mL of 5%glucose intravenous infusion and plus the treatment of control group, 1 to 10 days for a course.Patients of two groups were received 2 courses of treatment.The clinical efficacy, the incidence of leukopenia, white blood cell count to improve the rate and quality of life before and after treatment, and adverse drug reactions in two groups were compared.Results After treatment, total effective rate in treatment group was significantly higher than that in control group (84.91%vs 69.81%, P<0.05).After treatment 7,10 d, leukopenia rates in treatment group were 9.43%and 11.32%, significantly lower than 37.74%and 30.19%in control group (P<0.05).After treatment, the white blood cell count in the two groups all decreased than those before treatment, and the degree of decline in treatment group was higher than that in control group (P<0.05).After treatment, the rate of improvement of quality of life in treatment group was significantly higher than that in control group (88.68%vs 54.72%, P<0.05).The incidence of adverse drug reactions in treatment group was significantly lower than that in control group ( P<0.05).Conclusion Shenfu injection has a definitive clinical efficacy for the treatment of chemotherapy-induced leukopenia, which may improve immune function and the quality of life, worthy of further promotion in clinical practice.

BACKGROUNDSafe placement of the screws is a critical aspect of trans-pedicle internal fixation, and little information on in vivo morphology of the cervical vertebrae pedicle measured with imaging methods is available. The aim of this study was to measure the dimensions of cervical vertebrae C3 to C7 and provide screw length, screw diameter and tilt angle for clinical cervical vertebra trans-pedicle internal fixation.

METHODSThirty Chinese men and women underwent high-speed spiral computed tomography measurements to obtain data for C3 to C7, and the morphology of the cervical vertebra pedicles was reconstructed.

RESULTSReconstructed computer tomography image data revealed that: (1) pedicle sponge width increased incrementally from C3 to C7, (2) pedicle depth was similar for C3 to C7, (3) pedicle angle decreased incrementally from 47.20° to 33.76° for C3 to C7, and (4) pedicle point to midline distance was similar for C3 to C7. There were no statistical differences in morphological data between the right and the left side. Men had statistically larger values than women for all morphological parameters.

CONCLUSIONSReconstructed computed tomography images can provide useful data for clinical cervical vertebra trans-pedicle internal fixation. The individual measurement of cervical vertebra pedicles is recommended for safe placement of trans-vertebra pedicle screws.

Adult ; Bone Screws ; Cervical Vertebrae ; diagnostic imaging ; surgery ; Female ; Humans ; Image Processing, Computer-Assisted ; Internal Fixators ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Young Adult

Estrogen-related receptor alpha (ERRα) plays an important role in the development of hormone-dependent cancers, but its roles in lung cancer remain elusive. The present study was aimed to investigate the effects of ERRα on the proliferation and metastasis of lung cancer A549 cells. The mRNA and protein levels of ERRα were detected in lung cancer A549 and MCF-7 cells and bronchial epithelial BEAS-2B cells by qRT-PCR and Western blotting, respectively. ERRα plasmid transfection and XCT-790 (an inverse agonist of ERRα) were used to up-regulate or down-regulate ERRα expression in A549 cells, respectively. The viability of A549 cells was measured by cell counting kit-8 (CCK-8) and the motility of A549 cells by wound healing assay and Transwell migration/invasion assay. The epithelial markers E-cadherin (E-Cad) and zona occludin-1 (ZO-1), the mesenchymal markers fibronectin (FN) and vimentin (Vim) and the transcription factors (Snail, Zeb1 Twist and Slug) were further detected at mRNA and protein levels by qRT-PCR and Western blotting, respectively. The results showed that ERRα promoted the growth of lung cancer A549 cells in vitro. XCT-790 significantly inhibited the migration and invasion of A549 cells. Over-expression of ERRα promoted the epithelial-to-mesenchymal transition (EMT) of A549 cells, down-regulated the epithelial makers E-Cad and ZO-1, and up-regulated the mesenchymal makers FN and Vim. Silencing of Slug, but not other transcription factors, significantly abolished the ERRα-induced EMT of A549 cells. It was suggested that ERRα promoted the migration and invasion of A549 cells by inducing EMT, and Slug was involved in the process. Targeting ERRα might be an efficient approach for lung cancer treatment.
Cell Line, Tumor ; Cell Proliferation ; Epithelial-Mesenchymal Transition ; Gene Expression Regulation ; Humans ; Lung Neoplasms ; genetics ; metabolism ; prevention & control ; Neoplasm Metastasis ; Neoplasm Proteins ; biosynthesis ; genetics ; Receptors, Estrogen ; biosynthesis ; genetics