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.

OBJECTIVETo develvp a RP-HPLC method for the determination of flavonoids in fifteen kinds of flowers such as Iris lacteal pall, prunus persica and rosa chinensis.

METHODThe contents of quercetin, kaempferol and isorhamntin in fifteen kinds of flowers were extracted with methanol. The analysis was performed on a Kromasil C18 column (4.6 mm x250 mm, 5 microm) with methanol-0.1% phosphoric acid (50:50) as mobile phase.

RESULTThe quercetin, kaempferol and isorhamntin were separated well, and the result shows that the content of quercetin in the Iris lactea Pall was the highest (1.536%), the contene of kaempferol in Persica persice was the highest (0.572%), and the content of isorhamntin in chrysamthemum morifolium was up to 0.290%.

CONCLUSIONThe contents of flavonoids in these flowers were by determined RP-HPLC for the first time and the method can be used for quantitative determination of flavonoids in the flowers.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; chemistry ; Flowers ; chemistry ; Iris Plant ; chemistry ; Kaempferols ; chemistry ; Prunus ; chemistry ; Quercetin ; chemistry ; Rosa ; chemistry

Objective To investigate the application of the cross-sectional area ratio of internal jugular vein and common carotid artery (IJV/CCA) in the evaluating the volume responsiveness of critically ill patients. Methods The capacity of critically ill patients were prospectively assessed. The diameter and sectional area of the IJV and CCA were measured by bedside ultrasonography. The cross-sectional area ratio of IJV/CCA was calculated and compared with the variety of cardiac output (ΔCO) after passive leg raising (PLR). Then the correlation index between the cross-sectional area ratio of IJV/CCA and ΔCO was evaluated, and the sensitivity and specificity parameters of capacity status were assessed by the cross-sectional area ratio of IJV/CCA. Results Of 55 critically ill patients in this study, 34 cases had positive volume responsiveness, and 21 case negative volume responsiveness.The general clinical data of the two groups had no statistically significant difference. The cross-sectional area ratio of IJV/CCA in the positive group was significantly less than that of the negative group (1.38±0.55 vs. 2.16±0.68, P<0.01). There was a significant correlation between the IJV/CCA cross-sectional area ratio and the ΔCO value of PLR (r=-0.67, P<0.01). When the ratio of the cross-sectional area of IJV/CCA was 1.65, the sensitivity of the assessment capacity was 86.4％ and the specificity was 78.8％. Conclusions The use of portable bedside ultrasonography is a noninvasive, convenient and reliable method to evaluate the capacity state of the critically ill 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

To establish reference intervals for thyroid functional indicators in early (T1), mid-term (T2), and late stage (T3) pregnancy in a population of women in Northwestern China. A cross-sectional study was conducted on 620 pregnant women. Subjects were recruited through a questionnaire where apparently healthy women were selected. Serum thyroid stimulating hormone (TSH3), total triiodothyronine (TT3), total thyroid hormone (TT4), free triiodothyronine (FT3), and free thyroid hormone (FT4) were detected using the Beckman Unicel DXI 800 automatic chemiluminescence analyzer (the third-generation TSH detection reagent for TSH3),and the reference intervals of different gestation periods were established. The results showed that the reference intervals of TSH3 in T1, T2, and T3 were 0.05-4.59, 0.61-6.01, and 0.63-4.78 mIU/L, respectively; TT3 were 1.62-2.97 nmol/L, 1.59-2.95 nmol/L, and 1.45-2.70 nmol/L, respectively; TT4 were 95.49-185.00 nmol/L, 92.70-181.54 nmol/L, and 77.93-155.09 nmol/L, respectively; FT3 were 3.18-5.22 pmol/L, 2.78-4.67 pmol/L, and 2.51-4.18 pmol/L, respectively; and FT4 were 7.72-12.97 pmol/L, 6.90-1.09 pmol/L, and 5.63-9.85 pmol/L, respectively. All thyroid function indexes had statistically significant differences between the three stages of pregnancy (TSH: H=30.879, P<0.01;FT3: H =153.827, P<0.01;FT4: H =229.967, P<0.01;TT3: H =36.484, P<0.01;TT4: H =58.531, P<0.01). 20 independent samples were collected to verify the reference intervals of TSH, FT3, FT4, TT3 and TT4 for three trimesters of pregnancy, and all of them passed.

To establish reference intervals for thyroid functional indicators in early (T1), mid-term (T2), and late stage (T3) pregnancy in a population of women in Northwestern China. A cross-sectional study was conducted on 620 pregnant women. Subjects were recruited through a questionnaire where apparently healthy women were selected. Serum thyroid stimulating hormone (TSH3), total triiodothyronine (TT3), total thyroid hormone (TT4), free triiodothyronine (FT3), and free thyroid hormone (FT4) were detected using the Beckman Unicel DXI 800 automatic chemiluminescence analyzer (the third-generation TSH detection reagent for TSH3),and the reference intervals of different gestation periods were established. The results showed that the reference intervals of TSH3 in T1, T2, and T3 were 0.05-4.59, 0.61-6.01, and 0.63-4.78 mIU/L, respectively; TT3 were 1.62-2.97 nmol/L, 1.59-2.95 nmol/L, and 1.45-2.70 nmol/L, respectively; TT4 were 95.49-185.00 nmol/L, 92.70-181.54 nmol/L, and 77.93-155.09 nmol/L, respectively; FT3 were 3.18-5.22 pmol/L, 2.78-4.67 pmol/L, and 2.51-4.18 pmol/L, respectively; and FT4 were 7.72-12.97 pmol/L, 6.90-1.09 pmol/L, and 5.63-9.85 pmol/L, respectively. All thyroid function indexes had statistically significant differences between the three stages of pregnancy (TSH: H=30.879, P<0.01;FT3: H =153.827, P<0.01;FT4: H =229.967, P<0.01;TT3: H =36.484, P<0.01;TT4: H =58.531, P<0.01). 20 independent samples were collected to verify the reference intervals of TSH, FT3, FT4, TT3 and TT4 for three trimesters of pregnancy, and all of them passed.

Background::Hyperthermia in combination with DnaJA4-knockout (KO) obviously affects the anti-viral immunity of HaCaT cells. The mechanisms of this process are not yet fully explored. However, it is known that DnaJA4 interacts with actin cytoskeleton after hyperthermia. Our aim was to investigate the effects of DnaJA4 on F-actin in HaCaT cells following hyperthermia.Methods::Wild-type (WT) and DnaJA4-KO HaCaT cells were isolated at either 37°C (unheated) or 44°C (hyperthermia) for 30 min followed by testing under conditions of 37°C and assessing at 6, 12, and 24 h after hyperthermia. The cytoskeleton was observed with immunofluorescence. Flow cytometry and Western blotting were used to detect the expression of F-actin and relevant pathway protein.Results::DnaJA4-KO and hyperthermia changed the cytoskeleton morphology of HaCaT cells. F-actin expression levels were elevated in DnaJA4-KO cells compared with WT cells (6364.33 ± 989.10 vs. 4272.67 ± 918.50, P < 0.05). In response to hyperthermia, F-actin expression levels of both WT and DnaJA4-KO cells showed a tendency to decrease followed by an obvious recovery after hyperthermia (WT cells: unheated vs. 6 h after hyperthermia or 24 h after hyperthermia: 0.34 ± 0.02 vs. 0.24 ± 0.01, 0.31 ± 0.01, P < 0.001, P < 0.05; DnaJA4-KO cells: unheated vs. 6 h after hyperthermia or 24 h after hyperthermia: 0.44 ± 0.01 vs. 0.30 ± 0.01, 0.51 ± 0.02, P < 0.001, P < 0.01). WT cells restored to baseline levels observed in the unheated condition, while DnaJA4-KO cells exceeded baseline levels in the recovery. As the upstream factors of F-actin, a similar profile in rho-associated serine/threonine kinase 1 (ROCK 1) and RhoA expressions was observed after hyperthermia. While E-cadherin expression was decreased in response to hyperthermia, it was increased in DnaJA4-KO cells compared with WT cells. Conclusions::Hyperthermia affects the expression levels of F-actin in HaCaT cells. DnaJA4 knockout increases the expression of F-actin in HaCaT cells after hyperthermia. DnaJA4 regulates the expressions of F-actin and the related pathway proteins in response to hyperthermia in HaCaT cells.

Mercury sphygmomanometer based on traditional auscultation method is widely used in primary medical institutions in China, but a large amount of blood pressure data can not be directly recorded and applied in scientific research analysis, meanwhile auscultation data is the clinical standard to verify the accuracy of non-invasive electronic sphygmomanometer. Focusing on this, we designed a miniature non-invasive blood pressure measurement and verification system, which can assist doctors to record blood pressure data automatically during the process of auscultation. Through the data playback function,the software of this system can evaluate and verify the blood pressure algorithm of oscillographic method, and then continuously modify the algorithm to improve the measurement accuracy. This study introduces the hardware selection and software design process in detail. The test results show that the system meets the requirements of relevant standards and has a good application prospect.
Auscultation ; Blood Pressure/physiology* ; Blood Pressure Determination ; Oscillometry ; Sphygmomanometers

Objective: To investigate the risk factors of chronic obstructive pulmonary disease (COPD) in Li and Han ethnic group in Hainan, China. Methods: All subjects were randomly selected from various regions in Hainan. General characteristics were compared between COPD cases and healthy control cases in both Li and Han ethnic groups. The odds ratio (OR), the corresponding 95% confidence interval (CI) of COPD were calculated by logistic regression. Results: A total of 277 Li COPD cases, 307 Li healthy control subjects, 290 Han COPD cases and 301 Han healthy control were included in this study. In both the Li and Han groups, the average age exceeded 65 years, and the cigarette number smoked per day and the smoking duration were correlated with risk of COPD. In the Li COPD subjects, low weight, smoking, and recurrent infection of respiratory tract were mainly risk factors; while the mainly risk factor of Han COPD subjects was family history of respiratory disease. Conclusions: The risk factors are different in COPD subjects of Han and Li nationalities in Hainan of China. The age and smoking are strongly correlated with COPD risk.

To solve the problem of real-time detection and removal of EEG signal noise in anesthesia depth monitoring, we proposed an adaptive EEG signal noise detection and removal method. This method uses discrete wavelet transform to extract the low-frequency energy and high-frequency energy of a segment of EEG signals, and sets two sets of thresholds for the low-frequency band and high-frequency band of the EEG signal. These two sets of thresholds can be updated adaptively according to the energy situation of the most recent EEG signal. Finally, we judge the level of signal interference according to the range of low-frequency energy and high-frequency energy, and perform corresponding denoising processing. The results show that the method can more accurately detect and remove the noise interference in the EEG signal, and improve the stability of the calculated characteristic parameters.
Algorithms ; Electroencephalography ; Signal Processing, Computer-Assisted ; Signal-To-Noise Ratio ; Wavelet Analysis