Search

In this study, the recombinant plasmid pETDuet-INO1 was constructed by introducing the expression gene INO1 of inositol-1-phosphate synthase (IPS) (a key enzyme in inositol synthesis pathway) cloned from Saccharomyces cerevisiae SC288 by gene engineering technology into plasmid pETDuet-1. The recombinant strain Escherichia coli BL21/pETDuet-INO1 was constructed by introducing the recombinant plasmid pETDuet- INO1 into E. coli BL21 to achieve efficient expression of IPS in E. coli. The fermentation conditions for myo-inositol production were optimized by single factor experiments and orthogonal experiments. The results showed that the recombinant E. coli BL21/pETDuet-INO1 was successfully constructed, and the optimal fermentation conditions for myo-inositol production were as follows: initial glucose addition 12 g/L, induction temperature 28 ℃, and inoculum 3%. The yield of myo-inositol was (0.81±0.007) g/L after induction 24 h under these conditions, which was 60.39% higher than that before optimization.

To understand rapeseed (Brassica napus L.) heterosis on salt tolerance, hybrid cultivar Ningza 21 was used to investigate genes differentially expression compared with its parental lines. Through the next-generation high-throughput transcriptome sequencing technology with no salt treatments as negative control, 1078 differentially expressed genes (DEGs) were obtained, which were recognized to be closely related to heterosis of salt tolerance in oilseed rape at early seedling stage. These DEGs were annotated by released salt stress related QTLs, and expression of some DEGs were verified by qPCR, indicating the reliability of these DEGs. Additionally, the hybrid had more up-regulation candidate DEGs in shoot than those in root, and the expression pattern of DEGs was much closer to that of the maternal line than the paternal line in hybrid Ningza 21. More DEGs in hybrid similar to female parent showed positive regulation, and their function tended to be assimilation of biosynthesis. In contrast, DEGs similar to male parent were much related to redox and other regulatory functions. It was predicted that SNP loci were specific for heterosis of salt tolerance. All the results were expected to be helpful for salt tolerance improvement of hybrid rapeseed. [ABSTRACT FROM AUTHOR]

Improvement of pod shatter resistance is the basic target for rapeseed breeding suitable for mechanized harvesting. To better understand the inheritance of pod shatter resistance in Brassica napus, a doubled haploid (DH) population was used in the study. Two parents of the population had contrasting pod shatter resistance levels. Pod shatter resistance index was determined using random impact test method in two years, and the inheritance was analyzed by major gene plus polygene genetic model for plant quantitative traits as well as the analysis of skewness and kurtosis. Results showed that the pod shatter resistance was controlled by 3 genes with major effects and 5 - 7 polygenes with minor effects. There were major genes additive effects and complementary gene interactions. The heritability of major genes was greater than 85. 00% in both years, indicating pod shatter resistance was mainly controlled by major genes and less influenced by environments. Therefore, in breeding process for pod shatter resistance of rapeseed, pyramiding major genes from lines containing different major genes by hybridization and selection in early generation should be considered, ad the influence of environmental factors can be ignored. [ABSTRACT FROM AUTHOR]

The high-resolution remote sensing imagery such as Quick Bird (QB) and IKONOS are important imagery resources of Google Earth (GE). After processed graphically, such imagery are provided free of charge to the public to browse and use. In this study, both pixel-based and object-based classification methods were used to classify land-use classes respectively according to the GE imagery and the homologous QB imagery of the same area and time. Then, we compared the classification results of these two kinds of imagery in different land use classes and different classification methods. Based on the results gained above, we exponded the feasibility of using GE imagery in land-use classification and provided corresponding recommendations for the adaptability of using GE imagery in different situation of land use class and different classification methods. [ABSTRACT FROM AUTHOR]

The cDNA fragment of a putative pod shattering gene BnRPL was cloned from Brassica napus line R2 (easy to disperse seed) by RACE according to conserved region of AtRPL gene. Results showed that two BnRPLs were amplified named as BnRPL. a [with an open reading frame (ORF) of 1 758bp, coding 585 amino acids] and BnRPL. c (with an ORF of 1 734bp, coding 577 amino acids). Each BnRPLs included 3 introns and 4 exons. BnRPL. a and BnRPL. c was 92% identical. Comparing to RPL sequences from other species, BnRPL showed high homologous to AtRPL from Arabidopsis thaliana and RPL from Lepidium campestre. Variation of RPL protein occurred during the divergence from dicots and monocots. RT - PCR expression analysis showed that BnRPL gene was highly expressed in developing siliques(2Od after blooming). Moreover, it expressed in maturing silique wall and pseudoseptum, but not in seeds. [ABSTRACT FROM AUTHOR]

To further promote the use of recycled concrete in practice, Pseudo-static tests were carried out for 4 natural RC's ( reinforced concrete) columns and 8 recycled RC's columns. During the institute of building type test, the Load ( P) -displacement (Δ) relationship curves to each specimen were obtained, and the breakage mode, hysteresis loop, skeleton curves, energy dissipation, stiffness retrogression, ductility performance and carrying capacity of each specimen were similar to those of the natural RC's columns. In small axial ratio, the breakage mode was brittleness default, and in large axial ratio, the mode was ductility breakage. The ductility performance of recycled concrete is poor to the natural concrete. The fly ash could improve the ductility performance but lower the carrying capacity. In conclusion, the seismic behavior of recycled aggregate RC's column is apparently similar to that of natural RC's column. [ABSTRACT FROM AUTHOR]

According to pentatricopeptide repeat (PPR) protein sequence encoded by restorer gene from cytoplasmic male sterile rapeseed plant, a pair of degenerate primers was obtained after PCR screening of Nsa CMS (hybrid of Brassica napus and Sinapis arvensis). PCR results showed expected products specific to fertility restored plants and S. arvensis parent. The fragment was 309bp which was 69% identical to fertility restorer gene of Radish CMS, 70% identical to PPR gene cluster from the first chromosome of Arabidopsis thaliana, and 85% identical to a B. rapa accession clone which restores the fertility of pol CMS. Further analysis revealed that the amino acid sequence encoded by this fragment contained two adjacent PPR motifs with 35 amino acids each. Results indicated this fragment could be part of the candidate Rƒ gene for Nsa CMS. [ABSTRACT FROM AUTHOR]

Genetic diversity of 46 Brassica napus L. materials (40 have oil content over 43% and 6 below 40%)was analyz edusing SSR, SRAP and ST markers. 192 polymorphic bands were amplified by 81 primer airs. The allelic polymorphism information content (PIC) of those primers varied from 0.122 to 0.943, with an average of 0.512. The genetic distance of the analyzed materials ranged from 0.016 to 0.662, with an average of 0.409. The exotic varieties were more distant than the native ones. For example, the genetic distances were greater than 0.6 between Hektor and 7 other lines, and were greater than 0.5 between Sollux and 15 other lines. Cluste-ring analysis based on similarity coefficient showed that these 46 varieties could be divided into 5,9 and 18 groups, respectively at the point of 0.66,0.70 and 0.73.These results showed large genetic differences within the high oil content varieties, and indicated that the high oil content materials might possess different loci for in creasing oil content. Therefore, higher oil content could be expected in the future by pyramiding such non-allelic loci from these materials. [ABSTRACT FROM AUTHOR]

To promote the use of recycled concrete in practical engineering, pull -- out tests were carried out to study the influence of Coarse/Fine Recycled Concrete Aggregate (CRCA/FRCA) replacement rate, water -- cement ratio and the anchoring conditions on the compressive strength and bonding properties of Recycled Aggregate Concrete(RAC). The results show that: the bond strength of RAC rises with the increasing of CRCA replacement rate, and arrives at the maximum when the rate = 60%, then reduces; the bond strength of RAC reduces while the FRCA replacement rate increases, and it is better than that of ordinary concrete; too much CRCA/FRCA replacement will reduce the compressive strength of RAC; lower water -- cement ratio can improve the compressive strength and bond strength of RAC, and water -- cement ratio variation has less influence on RAC than that of ordinary concrete; appropriate increase in the thickness of concrete cover (≤3 d) and the anchorage length can increase the bond strength of RAC. In addition, by concerning the relationship between the bond strength of RAC and CRCA!FRCA replacement rate, water -- cement ratio, compressive strength, anchorage length, a formula was established. [ABSTRACT FROM AUTHOR]

Five self-compacting concrete beams and two corresponding normal concrete beams were designed to study the bearing properties of flexural members of self-compacting concrete, and their cracking loads, utmost loads, average strain in different heights of section, rigidness, and crack were investigated by experiment. And on the basis of methods to calculate common concrete members, the formulas to calculate the rigidity, crack and ultimate height of compression strain of SCC as well as the method to calculate the bearing capacity were provided. [ABSTRACT FROM AUTHOR]

In order to investigate mechanical properties of partial recycled concrete beams, 15 partial recycled concrete beams with 5 kinds of reinforcement ratios were studied by experiment. The results indicated that the plain section presumption could be applied to partial recycled concrete structures. The bearing capacity of normal section for partial recycled concrete flexural member was not lowered compared with that of ordinary concrete flexural member, but the stiffness of partial recycled concrete flexural member was degraded. The calculating formula of flexural bearing capacity and the modified stiffness calculating formula of partial recycled concrete flexural member were established. The suggested values of ultimate compression strain of axial compression and eccentricity compression, as well as the minimum reinforcement ratio were provided. Finite element analysis of partial recycled concrete flexural member was performed by ANSYS. The calculation results were in good agreement with experimental results. [ABSTRACT FROM AUTHOR]

Objective: To observe the clinical effect of moxibustion therapy based on "sancai yizhi" (benefiting the intelligence) therapy on the improvements of memory function and serum protein markers, Aβ1-42, Tau and phosphorylated Tau （P-tau） in the patients with amnestic mild cognitive impairment (aMCI), and has a preliminary exploration on its peripheral mechanism., Methods: A total of 120 patients with aMCI were divided into a moxibustion group and a medication group using a random number table, 60 patients in each group. In the moxibustion group, 6 cases were dropped out and 5 cases were withdrawn, and then 49 cases accomplished the trial finally. In the medication group, 8 cases were dropped out and 6 ceases were withdrawn, thus 46 cases finally accomplished the trial. In the moxibustion group, moxibustion therapy was provided at Baihui (GV20), Shenque (CV8) and bilateral Yongquan (KI1), once every other day, 20 minutes each time, totally for 8 weeks. In the medication group, donepezil hydrochloride tablets were administered orally, 5 mg once a day, consecutively for 8 weeks. The scores of Rivermead behavioral memory test (RBMT) and Monterey cognitive assessment (MoCA) scale were adopted as the indicators to evaluate the therapeutic effect after treatment in the two groups. Enzyme linked immunosorbent assay (ELISA) was used to detect the changes of the levels of serum protein marker levels, i.e. Aβ1-42, Tau and P-tau before and after treatment in the patients of two groups., Results: Compared with the scores before treatment, RBMT score and MoCA score all increased after treatment in the patients of two groups ( P <0.05). Compared with the medication group at the same time points, RBMT score increased significantly ( P <0.05) in the moxibustion group after treatment. In the moxibustion group, as compared with the levels before treatment, the levels of serum Aβ1-42,Tau and P-tau decreased after treatment in the patients ( P <0.05). But in the medication group, the levels of serum Aβ1-42 and P-tau were reduced ( P <0.05). Compared with the medication group at the same time points, there were no significant differences in the changes of serum Aβ1-42,Tau and P-tau in the moxibustion group ( P >0.05)., Conclusion: Moxibustion therapy based on "sancai yizhi" theory improves the cognitive function in the patients with aMCI and it affects the levels of serum Aβ1-42, Tau and P-tau, which may be the reason for the improvement of cognitive function in the patients with aMCI.

Objective: To investigate the clinical effect and safety of moxibustion therapy based on Sancai theory for reinforcing intelligence in the early intervention of mild cognitive impairment (MCI)., Methods: A total of 210 patients with MCI were divided into moxibustion group and medication group using a random number table, with 105 patients in each group. The patients in the moxibustion group were given moxibustion at Baihui (GV20), Shenque (CV8), and Yongquan (KI1) once every other day, 20 minutes each time, for a total of 8 weeks, and those in the medication group were given oral Nimodipine 30 mg once a day for 8 weeks. Mini-Mental State Examination (MMSE) score, score of meaningless graphics recognition of Clinical Memory Scale (CMS), and event-related potential P300 latency were evaluated before treatment, after 8 weeks of treatment, and at 12 weeks after treatment, and the safety of treatment was analyzed for both groups., Results: Both groups had significant increases in MMSE score and the score of meaningless graphics recognition of CMS and a significant reduction in P300 latency after treatment and during follow-up ( P <0.01, P <0.05). Compared with the medication group, the moxibustion group had a significant increase in MMSE score and a significant reduction in P300 latency after treatment ( P <0.05)., Conclusion: Moxibustion therapy based on Sancai theory for reinforcing intelligence can improve cognitive impairment in patients with MCI and is thus an effective intervention method for improving the cognitive function of patients with MCI.

Objective: To evaluate the correlation of Fast-track extubation ultrasound score (FTEUS) and clinical multi-organ information indicators in post-cardiac surgery patients., Methods: prospectively recruit post-cardiac surgery patients who were about to extubating from Febuary 2019 to September 2019. A fast-track extubation ultrasound score protocol (FTE-USP) was developed on the basis of the conventional fast-track extubation standard precisely and individualized. Cardiac, pulmonary and inferior vena cava ultrasound examinations were performed by specially trained observers, video data were saved, FTE-USP was used for scoring, Kendall consistency coefficient was used to meature the interobserver consistency. The correlation between the FTEUS and the patients' clinical indicators was evaluated., Results: A total of 207 patients were recruited in the study, including 89 males and 118 females, aged (54.63±11.80) years. The FTEUS was performed at bedside with a mean time of (8.23±2.08) min, Kendall consistency coefficient is 0.941. With the increase of the total score of FTEUS, the incidence of clinical adverse events increased (especially the arrhythmia), and there were significant changes in liver, kidney, heart, lung and other organ function indicators, among which serum creatinine level, serum cystatin C level, serum NT-pro-brain natriuretic peptide, length of stay in intensive care unit, non-invasive mechanical ventilation time after extubation, and incidence of arrhythmia were positively correlated with FTEUS ( P < 0.05).With FTEUS increased to 5 points, the incidence of arrhythmia (14/24, 58.3%), cardiopulmonary resuscitation (2/24, 8.3%) and weaning failure (2/24, 8.3%) increased., Conclusion: FTE-USP integrates multi-organ informations, can be performed quickly at the bedside and alerts adverse events. It has the potential to be applied to assist clinical decision-making in post-cardiac surgery patients before extubation., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).)

Objective To prepare a lentiviral vector expressing LLGL2 and establish KYSE450 and TE-1 cell lines for the stable expression of LLGL2. Methods The full-length LLGL2 sequence was amplified by high-fidelity PCR, and then it was inserted into pCDH-CMV-IRES-GFP-EF1-Puro vectors. The recombinant plasmid was confirmed by double enzyme digestion and sequencing. After co-infection of pCDH-CMV-LLGL2-IRES- GFP-EF1-Puro with vesicular stomatitis virus glycoprotein (VSVG) and PHR into HEK293T cells, the lentivirus was harvested and used for infecting esophageal squamous cell carcinoma cell lines including KYSE450 and TE-1 cells. These two cell lines infected with the lentivirus were screened with puromycin, and the stable cell lines were further confirmed with green fluoresence and Western blotting. Results Dual-enzyme digestion and sequencing confirmed that the pCDH-CMV-LLGL2-IRES-GFP-EF1-Puro vector, a lentiviral expression vector for the overexpression of LLGL2, was successfully constructed through high-fidelity PCR and ligation. Western blotting showed the increased expression level of LLGL2 protein in KYSE450 and TE-1 stable cell lines compared with the controls. Conclusion The experiment successfully established KYSE450 and TE-1 stable cell lines for the overexpression of LLGL2.

Objective: To investigate sexual satisfaction (SS) and the factors associated with decreased SS among individuals with hearing disability., Methods: We conducted an investigation on SS among 439 individuals (268 males and 171 females, aged ≥18 yr) with hearing disability using a general information questionnaire, Center for Epidemiologic Studies Depression Scale, Social Support Rating Scale, and a self-report on SS. We identified the factors of decreased SS by multivariate ordinal logistic regression analysis., Results: Totally 76 (17.3%) of the hearing-disability individuals investigated were dissatisfied with their sexual life. SS reduction was significantly correlated with the status of being single (OR=1.72), grade-1 or -2 disability (OR=1.78), physical diseases (OR=2.46), depression (OR=6.61), or inadequate subjective social support (OR=3.28)., Conclusions: SS of hearing-disability persons is relatively low, which can be improved by treating physical diseases, promoting mental health, and providing psycho-social support.

Objective: Construction and identification of recombinant adenovirus vector with human/mouse interferon (IFN-gamma) and effectively transfection into human umbilical cord mesenchymal stem cells (HUMSCs)., Methods: IFN-gamma gene of human/mouse were amplified from the plasmid by polymerase chain reaction (PCR) and then inserted into the plasmid pDC316 to generate pDC316-IFN-gamma. After being confirmed by restriction enzyme digestion and DNA sequencing, the DNA encoding IFN-gamma in the new structure were inserted into the vector of recombinant plasmid adenovirus and confirmed by restricition enzyme digestion. Then the human embryonic kidney cell line 293 were transfected with confirmed Ad-IFN-gamma, and the recombinant adenovirus were amplified, and the virus titer were detected using 50% tissue culture infective dose (TCID50) assay. The expression of the green fluorescent protein (GFP) and IFN-gamma were detected by fluorescent microsope and Western blot and ELISA after the recombinant adenovirus transfected HUMSCs., Results: The recombinant adenovirus Ad-hIFN-gamma were constructed successfully, and amplified with titer of 1.6 x 10(10) IU/mL. The titer of Ad-mIFN-gamma was 1.0 x 10(10) IU/mL and the titer of Ad-GFP was 1.0 x 10(9) IU/mL. The green fluorescence proteins could be observed under fluorescent microscope in HUMSCs 24 h after transfection and with a stronger degree after 72 h, and IFN-gamma expression in HUMSCs were confirmed by Western blot and ELISA., Conclusion: Construction and identification of recombinant adenovirus vector of human/mouse IFN-ygamma and effectively transfection of HUMSCs were successful.

Objective: To explore the effect of land consolidation project on the detention of Oncomelania hupensis snails in the flood at the Yangtze River beach., Methods: The land consolidation project including removing wild willow groves, cutting the high and filling the low, and improving the slope was carried out at the Yangtze River beach in 2011, and the status of snails was investigated before and after the project, respectively, at the end of the flood season., Results: Before the project (2010), at the end of the flood season, the rate of the frames with snails was 96.5% and the snail density was 3.655/0.1 m2. After the project (2012), at the end of the flood season in the same area, the rate of the frames with snails was 2.0% and the snail density was 0.015/0.1 m2 (chi2 = 357.29, P < 0.01)., Conclusion: The land consolidation has a significant effect on the detention of snails in the flood at the Yangtze River beach.

Objective: To study the expression of beclin1 in renal tissue of diabetic rats, and its role in diabetic nephropathy (DN)., Methods: Diabetic rat model was produced by intraperitoneal injection of streptozotocin (50 mg/kg), it was considered to be diabetic nephropathy that 72 h blood glucose after modeling was higher than 16 mmol/L. The positive urine protein was detected by paper method at the end of 1 week. 24 h urine albumin excretion (ALb) and urinary albumin and urinary creatinine ratio (ACR) were examined by immunoturbidimetry, Beclin1 expression was also studied by immunohistochemistry with envision method. The electron microscopy was used to detect the form of autophagic vesicles and lysosome., Results: (1) Compared with control group, the 24-hour urinary albumin excretion rate and ACR in diabetic rats were significantly higher. (2) The electron microscopy showed there were autophagic vesicles in renal tissue in both groups; (3) Immunohistochemistry showed beclin1 expression in normal rat kidney cells partly, while no expression in glomeruli. The expression of beclin1 in DN renal tissue was stronger than that in normal rats (P < 0.05)., Conclusion: beclin1 may be involved in the pathogenesis of diabetic nephropathy with the mechanism related to the process of programmed cell death.

Histidines provide axial ligands to the primary electron donors in photosynthetic reaction centers (RCs) and play an important role in the protein environments of these donors. In this paper the authors present a systematic study of ionization energies and vibrational properties of histidine using hybrid density functional theory (DFT). All calculations were undertaken by using B3LYP method in combination with four basis sets: 6-31G(d), 6-31G(df, p), 6-31+G(d) and 6-311+G(2d, 2p) with the aim to investigate how the basis sets influence the calculation results. To investigate solvent effects and gain a detailed understanding of marker bands of histidine, the ionization energies of histidine and the vibrational frequencies of histidine which are unlabeled and 13C, 15N, and 2H labeled in the gas phase, CCl4, protein environment, THF and water solution, which span a wide range of dielectric constant, were also calculated. Our results showed that: (1) The main geometry parameters of histidine were impacted by basis sets and mediums, and C2-N3 and N3-C4 bond of imidazole ring of histidine side chain display the maximum bond lengths in the gas phase; (2) single point energies and frequencies calculated were decreased while ionization energies increased with the increasing level of basis sets and diffuse function applied in the same solvent; (3) with the same computational method, the higher the dielectric constant of the solvent used, the lower the ionization energy and vibrational frequency and the higher the intensity obtained. In addition, calculated ionization energy in the gas phase and marker bands of histidine as well as frequency shift upon 13C and 15N labeling at the computationally more expensive 6-311+G(2d, 2p) level are in good agreement with experimental observations available in literatures. All calculations indicated that the results calculated by using higher level basis set with diffuse function were more accurate and closer to the experimental value. In conclusion, the results provide useful information for the further studies of the functional and vibrational properties of chlorophyll-a ligated to histidine residue in photosynthetic reaction center.

Aim: To investigate the toxic response in auditory cortex of guinea pigs caused by cis-platinum (DDP), and the protective role of melatonin in this effect., Methods: Cis-platinum and melatonin were injected peritoneally. LDH, MDA, NO in the auditory cortex were detected by spectrophotometeR., Results: The body weight of the guinea pigs was diminished by peritoneal injection of Cis-platinum for 7 days (P < 0.01). Peritoneal injection of Cis-platinum induced the increased leakage of LDH (P < 0.05 vs injection of normal saline). This effect was reduced by injection of MT (P < 0.05). The content of MDA in the auditory cortex was also increased because of injection of Cis-platinumv for 7 days (P < 0.01) and MT reduced this effect (P < 0.05). The change of NO in the auditory cortex was not statistically significant after injection of Cis-platinum or Cis-platinum with MT., Conclusion: Peritoneal injection of Cis-platinum could destroy neurons in the auditory cortex. This effect could be reduced by melatonin by an anti-free radials mechanism.

The plant height of rapeseed varieties has increased more than 20 cm due to wide application of heterosis, which leads to high risk of lodging at late stages of rapeseed development. Using dwarf genes to decrease plant height is an effective approach to resolve the lodging problem. A dwarf mutation 99CDAM with plant height of about 85 cm was discovered from a Brassica napus line which had selfed for many years. The mutation 99CDAM has good characters of early flowering and rich branches, as well as better yield and quality traits, which can be stably inherited, so 99CDAM has important value in Brassica napus breeding. Genetic analysis on reciprocal crosses between 99CDAM with high-stalk lines 2091, 7045 and 7350, and the F2BC1 and F2:3 populations derived from the cross between 2091 with 99CDAM indicated that the genetic model of dwarf genes in 99CDAM was obviously different from what had been reported before. The mutation 99CDAM was controlled by three pairs of recessive dwarf genes and showed a maternal effect..

Objective: Despite the causes for melanin increase, the increased gene expression of TYR is a common pathological process. Based on this viewpoint, antisense-S-Oligo of TYR was designed and synthesized to regulate synthesis of melanin in order to explore the treatment for skin pigmentation., Methods: The cultured melanocytes were divided into 3 groups. The group 1 was treated with endothelin, group 2 treated with ultraviolet ray and group 3 was used as the control. In each group, the 5' antisense-S-Oligo, the 3' antisense-S-Oligo, the mixed antisense-S-Oligo of TYR or Dotap only was added. The melanin content and TYR gene expressions were examined., Results: The 5' antisense-S-Oligo, the 3' antisense-S-Oligo and the mixed antisense-S-Oligo significantly inhibited the increase of melanin content and TYR gene expression, which were caused by endothelin or ultraviolet ray treatment. Of the three treatments, the 3' antisense-S-Oligo showed the strongest effect., Conclusion: Antisense-S-Oligo has significant regulating effects on TYR gene expression and melanin content. The 3' antisense-S-Oligo is more effective than the 5' antisense-S-Oligo.